Production, purification, characterization, immobilization, and application of β-galactosidase: a review...
ASIA-PACIFIC JOURNAL OF CHEMICAL ENGINEERING Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 Published online 22 April 2014 in Wiley Online Library (wileyonlinelibrary.com) DOI: 10.1002/apj.1801
Special theme review
Production, purification, characterization, immobilization, and application of β-galactosidase: a review Arijit Nath,1† Subhoshmita Mondal,1† Sudip Chakraborty,1,2* Chiranjib Bhattacharjee1 and Ranjana Chowdhury1 1 2
Chemical Engineering Department, Jadavpur University, Kolkata, West Bengal 700032, India Department of Chemical Engineering and Materials, University of Calabria, Cubo-44C, Rende 87036,CS, Italy
Received 20 June 2013; Revised 29 September 2013; Accepted 5 October 2013
ABSTRACT: Biopharmaceuticals are new categorizing biomolecules, which are the results of incredible proliferation in the field of biotechnology. One of the challenging biomolecule β-galactosidase (β-galactosidase galacto hydrolysase, trivially lactase) catalyzes hydrolysis of lactose to produce glucose and galactose, and in some cases, it takes part in transgalactosylation reaction that produces functional food galato-oligosaccharide. A wide variety of strategies had been already attempted for production of β-galactosidase through fermentative route. Beside the upstream process, downstream technology towards purification and immobilization of target enzyme also create great attentions. Subsequently, its wide applications in the field of food, biopharmaceuticals, dairy, diagnosis, and waste treatment boost up biotechnological economy as well as zero-effluent discharge. In dairy industry, β-galactosidase has been used to degrade lactose, prevent crystallization of lactose, improve sweetness, and increase the solubility of milk product, otherwise which would be an environmental pollutant. In food and pharmaceutical industries, β-galactosidase has been used to prepare low lactosecontaining food products for low lactose-tolerant people. Therefore, it is obvious to elucidate different technological aspects of β-galactosidase, which may provide a great knowledge in educational and industrial field. Taking the enzyme into account, a ready review has been made about its production, purification, characterization, and immobilization technology. The review also addresses wide applications of β-galactosidase in different fields. © 2014 Curtin University of Technology and John Wiley & Sons, Ltd. Keywords: β-galactosidase; production; purification; characterization; immobilization; application
INTRODUCTION Emergence of biotechnology, which brings a boon to biological origin, possesses a challenging revolution to the categorical field, biopharmaceuticals. There is enormous range of therapeutics that involves biopharmaceuticals, and it combines biomolecular forms, which extensively give rise to the development of the microbial synthesis of diverse enzymes and metabolites. Intellectual revolution with new visions and hopes by dispensation of any method of bioprocess that deals with the design and development of equipments for the manufacturing of various products, such as food-beverages, sera, new medicines, semisynthetic organs, antibiotics, and enzymes from biological sources creates a great attention. This is responsible for the explosion of various biotechnological processes used in industries for large-scale production of *Correspondence to: Sudip Chakraborty, Chemical Engineering Department, Jadavpur University, Kolkata, West Bengal 700032, India. E-mail:
[email protected] † Authors have similar contribution. © 2014 Curtin University of Technology and John Wiley & Sons, Ltd. Curtin University is a trademark of Curtin University of Technology
biological products and optimization of their yields as well as the qualities of end products.[1] The lactose-hydrolyzing enzyme β-galactosidase has been accepted since long as an important ingredient in dairy, food processing, and pharmaceutical industries. β-galactosidase catalyzes hydrolysis of lactose into glucose and galactose and also takes part in transgalactosylation reaction that produces galatooligosaccharide (GOS) (e.g., Gal (β1 → 3) Gal (β1 → 4) Gal (β1 → 6)).[2,3] One of the most common disease hypolactasia (lactose intolerance) or lactose maldigestion is caused by insufficient synthesis of lactose-cleaving enzyme β-galactosidase in the brush border membrane of mucosa of the small intestine. In most cases, this causes several symptoms, which may include abdominal bloating and cramps, flatulence, diarrhea, nausea, rumbling stomach, or vomiting after consuming significant amounts of lactose.[4] In most of the human races, β-galactosidase is found to be lost during the first or second decade of life, and only some isolates are seen among people of Northern European origin and their overseas descendants. African and Indian communities maintain a high intestinal lactase
Asia-Pacific Journal of Chemical Engineering
activity throughout life. According to reviews of Scrimshaw, Murray, 1988 and Sahi, 1994, the global prevalence of lactose maldigestion are above 50% in South America, Africa, and Asia reaching almost 100% in some Asian countries such as China. In the USA, prevalence is 15% among Whites, 53% among Mexican-Americans, and 80% in the Black population. In Europe, it varies from around 2% in Scandinavia to about 70% in Sicily. Australia and New Zealand have prevalence of 6% and 9%, respectively.[5,6] β-galactosidase has been used in biopharmaceutical, food, and dairy industries to prevent crystallization of lactose, to improve sweetness, to increase the solubility of milk product, to prepare low lactose-containing food products for low lactose-tolerant people, and for the utilization of cheese whey, which would otherwise be an environmental pollutant.[7,8] Therefore, it is obvious to elucidate different technological aspects of β-galactosidase with special interest in production, purification, characterization, immobilization technology as well as its wide applications in different process industries. The present review may provide a great knowledge with fill-up gaps of educational field and industrial sector.
SYNTHESIS MECHANISM OF β-GALACTOSIDASE The control system of β-galactosidase synthesis was first worked out by Jacob and Monod in1961, at the molecular level. In prokaryotic type of gene expression, the lac operon showed inducible system with the control of enzymes that are produced in the presence of lactose.[9] Presences of lac operon in Escherichia coli and Bacillus sp. including lactic acid bacteria have potential of synthesizing intracellular β-galactosidase. Meanwhile, lactose metabolism in E. coli synthesizes several proteins, such as β-galactosidase, which converts lactose into glucose and galactose, β-galactoside permease that transports lactose into the cellular system,
β-GALACTOSIDASE: A REVIEW
and the function of β-galactoside transacetylase is yet to be known. The construction of lac operon is described in Fig. 1. For any consortia, all the genes involved in controlling this pathway are located next to each other on the chromosome, and together they form an operon. Generally, β-galactosidase synthesis has been often demonstrated considering E. coli as a model microorganism. The genetic switches can also combine positive and negative controls. The lac operon consists of three structural genes and a promoter, a terminator, regulator, and an operator. The three structural genes are lacZ, lacY, and lacA. Regulation for the specific control of the lac genes depends on the availability of the substrate lactose. The proteins are not produced by the bacterium when lactose is unavailable as a carbon source. The lac genes are organized into an operon oriented in the same direction, immediately adjacent on the chromosome and are co-transcribed into a single polycistronic mRNA molecule. The lac operon in E. coli, unlike trp operon, works under both negative and positive transcriptional controls by the lac repressor protein and catabolite activator protein (CAP), respectively. CAP enables bacteria to use alternative carbon sources such as lactose in the absence of glucose. The CAP cannot induce expression if lactose is not present, and the lac repressor ensures that the lac operon is shutoff in the absence of lactose. This arrangement enables the lac operon to respond to and integrate two different signals, so that it is expressed only when two conditions are met, i.e., lactose must be present and glucose must be absent. Transcription of all genes starts with the binding of the enzyme RNA polymerase (RNAP), a DNA-binding protein that binds to a specific DNA binding site, the promoter, immediately upstream of the genes. Binding of RNA polymerase to the promoter is aided by the cyclic adenosine monophosphate (cAMP)-bound CAP (also known as the cAMP receptor protein). From this position, RNAP proceeds to transcribe all three genes (lacZ, lacY, and lacA) into mRNA.[11]
Figure 1. Construction of lac operon (Figure adapted from K. L. Anderson, G. Purdom
(2008)).[10]
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
331
332
Asia-Pacific Journal of Chemical Engineering
A. NATH ET AL.
Figure 2. Regulation of lac operon in (A) presence and (B) absence of lactose.[12]
Control mechanism of lac operon In the absence of lactose, the regulatory response to lactose uses an intracellular regulatory protein, which is called the lactose repressor to hinder the synthesis of β-galactosidase. The lacI gene coding for the repressor lies nearby the lac operon and is always expressed (constitutive). If lactose is absent in the growth medium, the repressor binds very tightly to a short DNA sequence just downstream of the promoter near the beginning of lacZ called the lac operator. The repressor binding to the operator interferes with binding of RNAP to the promoter and therefore mRNA encoding LacZ and LacY are only made at very low levels. During the cells growth in the presence of lactose, a lactose metabolite allolactose, which is a combination of glucose and galactose, binds to the repressor, causing a change in its shape. Thus, the altered repressor is unable to bind to the operator, allowing RNAP to transcribe the lac genes and thereby leading to higher levels of the encoded proteins. The schematic diagrams of the proposed phenomena are described in Fig. 2. The second control mechanism is in response to glucose, which is transported into the cell by the phosphoenolpyruvate (PEP)-dependent phosphotransferase system. The phosphate group of PEP is transferred via a phosphorylation cascade, consisting of the general phosphotransferase system (PTS) proteins HPr and EIA, and the glucose-specific PTS proteins, EIIAGlc and © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
EIIBGlc, which are the cytoplasmic domain of the EII glucose transporter. Transport of glucose is accompanied by its phosphorylation by EIIBGlc through removing the phosphate group from the other PTS proteins, including EIIAGlc. The unphosphorylated form of EIIAGlc binds to the lac permease and prevents it from bringing lactose into the cell. Therefore, if both glucose and lactose are present, the transport of glucose blocks the transport of the inducer of the lac operon. This process is called inducer exclusion.[13] In other way, 16 base pairs upstream site of the promoter, which is used for a positive control of the gene expression is known as CAP site or cAMP protein site or catabolite gene activator (cga) site, because it is utilized for binding of CAP to encourage gene expression. CAP can bind to this site only when it is bound with cAMP. Hence, a positive control is exerted over the transcription process by the CAP–cAMP complex with an effect exactly opposite to that of repressor binding to an operator. Moreover, the effector molecule cAMP determines the effect of CAP on lac operon transcription in the presence of glucose, it inhibits the formation of cAMP and prevents it to bind to CAP. The DNA-bound CAP is then able to interact physically with RNA polymerase and essentially increase the affinity of RNA polymerase for the lac promoter. In this way, the catabolite repression system contributes to the selective activation of the lac operon.[14] The schematic diagram of mechanism of carbon catabolic repression is described in Fig. 3. Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering
β-GALACTOSIDASE: A REVIEW
Figure 3. Mechanism of carbon catabolic repression (A) inducer exclusion and (B) induction
prevention (Figure adapted from B. Görke and J. Stülke (2008)).[13]
A mechanism of carbon catabolite repression by which the activity of PTS-regulation domaincontaining transcription factors, which is inhibited in the presence of preferred carbon sources, is called Induction prevention. It proceeds with phosphorylation of histidine protein (HPr) at Ser46 by HPr kinase/ phosphorylase (HPrK). This phosphorylation occurs when the intracellular concentrations of fructose-1, 6-bisphosphate (FBP) and adenosine triphosphate are high, which reflects the presence of preferred carbon sources. HPr(Ser-P) binds to CcpA protein, and this interaction is enhanced by glycolytic intermediates, such as FBP and glucose-6-phosphate. The complex of CcpA and HPr(Ser-P) binds to cre sites on the DNA and thereby represses the transcription of catabolic genes. HPrK is also responsible for dephosphorylation of HPr(Ser-P) under conditions of high inorganic phosphate (Pi) and low adenosine triphosphate and when there is poor nutritional supply of FBP. Also, HPr(His-P) contributes to carbon catabolic repression: in the absence of glucose, HPr(His-P) phosphorylates glycerol kinase (GlpK), and transcriptional regulators that contain PEP–carbohydrate phosphotransferase system-regulatory domains, which acts as precursors for their activity. Thus, in the presence of glucose, activation of the phosphotransferase system-regulatory domain regulators by their inducers is prevented.[13] Research with this system is greatly added by the availability of constitutive mutants. A constitutive mutant of any consortia continuously produces gene products where there is no control over its expression. In these mutants, the aforementioned proteins are produced all the time in comparison to the wild type where the proteins only appear in the presence of lactose. So in these mutants, the mutation must be a gene other than those responsible for the structural gene.[15] © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
PRODUCTION OF β-GALACTOSIDASE β-galactosidase belongs to the group of saccharides converting enzymes, i.e., in the family of hydrolases. They are widespread, distributed in numerous biological systems, e.g., microorganisms (yeasts, fungi, bacteria, and actinomycetes), plants, and animal tissues. Compared with animal and plant sources, microbial-synthesized enzyme provides higher yields, which may decrease its production cost. Therefore, production of β-galactosidase through microbial route creates a great attention. Although, the most studied β-galactosidase is produced by E. coli, possible toxic factors associated with coliforms make it unlikely that crude isolates of this enzyme, which may be permitted in food processes.[16–21] In industrial scale, production of β-galactosidases is carried out using generally recognized as safe microorganism, yeast (mainly from Kluyveromyces marxians, Kluyveromyces lactis, and Kluyveromyces fragilis) and fungal (mainly from Aspergillus niger and Aspergillus oryzae) consortia. The detail works carried out in this direction have been represented in Table 1.
PURIFICATION OF β-GALACTOSIDASE Different separation techniques, such as membranebased separation, ion exchange membrane chromatography, gel permeation chromatography, zinc chloride, protamine sulfate, and ammonium sulfate precipitation; had already been attempted for purification of β-galactosidase from crude extract. The detail works carried out in this direction have been represented in Table 2. Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
333
Bacillus licheniformis ATCC 12759 Lactobacillus acidophilus NRRL 4495 Bifidobacterium animalis Bd12, Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842
Bifidobacterium animalis Bd12, Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 Lactobacillus plantarum Pi06 Lactobacillus acidophilus Streptococcus thermophilus Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842
Bacillus coagulans RCS3 Aspergillus flavus Penicillium chrysogenum NCAIM 00237 Aspergillus oryzae PTCC 5163
LacA from Aspergillus oryzae, expressing in Saccharomyces cerevisiae NCYC869-A3/ pVK1.1 Rhizomucor sp. Aspergillus niger
Bacteria Bacteria Bacteria
Bacteria
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Bacteria Fungus Fungus Fungus
Fungus
Lactose-based chemically defined medium Lactose-based yeast nitrogen base medium and semi-synthetic lactose medium
Lactose-based chemically defined medium Luria medium where individually galactose, glucose, mannitol, lactose, and xylose were added Luria medium Acid whey Chemically defined medium where individually glucose, lactose, and galactose were used. Moreover, individually peptone, yeast extract, casein hydrolysate, tryptone, or ammonium sulfate were used De-proteinated whey where protein was supplemented by individually peptone, yeast extract, casein hydrolysate, tryptone, and ammonium sulfate Lactose-based chemically defined medium de-Mann Rogosa and Sharpe medium Whey-based medium Skim milk, modified de-Mann Rogosa Sharpe medium, whey, and whey permeate-based broth were used. Additionally, yeast extract was added Lactose-based chemically defined medium Lactose-based chemically defined medium Lactose-based chemically defined medium Whey and chemically defined medium with different carbohydrate, i.e., lactose, wheat barn, and soybean meal Wheat bran and rice husk
Combination of skim milk powder, glucose, and yeast extract
Cheese whey Cheese whey Supplemented whey with cauliflower waste Lactose-based chemically defined medium Whey Lactose-based chemically defined medium Whey Lactose-based chemically defined medium Lactose-based yeast-defined mineral medium Lactose-based chemically defined medium, de-proteinated whey Whey Basal medium with IPTG
Substrate
Batch Continuous
Batch
Batch Batch Batch Batch
Batch Batch Batch Batch
Batch
Batch Batch Batch
Batch Batch
Batch
Fed batch Batch Batch Batch Batch Batch Batch Batch and fed batch Batch Fed batch Batch Batch
Operating mode of bioreactor
[44] [45]
[43]
[41] [42]
[39] [40]
[21]
[20] [38]
[19]
[18]
[17]
[37] [16]
[35] [36]
[34]
[32] [33]
[29] [30] [31]
[26] [27] [28]
[24] [25]
[22] [23]
Reference
A. NATH ET AL.
Fungus Fungus
Bacteria Bacteria Bacteria Bacteria
Bacteria Bacteria
Bacteria
Kluyveromyces marxians Kluyveromyces marxians Kluyveromyces marxians Kluyveromyces marxians CBS 7894 Kluyveromyces marxians NCIM 3551 Kluyveromyces marxians CCT 7802 Kluyveromyces marxians MTCC 1388 Kluyveromyces fragilis NRRL-Y-1109 Recombinant Saccharomyces cerevisiae W303 Saccharomyces cerevisiae Candida pseudotropicallis Cloned Escherichia coli XL-1 Blue, XL-1 Blue FF, TB-1 and N4830 Lactobacillus sp. Bulgaricus CHR Hansen Lb-12 Bacillus sp. MPTK 121 Bacillus sp.
Name of consortium
Yeast Yeast Yeast Yeast Yeast Yeast Yeast Yeast Yeast Yeast Yeast Bacteria
Type of consortium
Table 1. Different types of consortium used in β-galactosidase production.
334 Asia-Pacific Journal of Chemical Engineering
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Ammonium sulfate precipitation, hydrophobic interaction chromatography, and affinity chromatography Gel permeation chromatography, affinity chromatography, and ammonium sulfate precipitation Column chromatographies on resource Q and sephacryl S-200 HR Ammonium sulfate precipitation and gel permeation chromatography Protamine sulfate precipitation, ammonium sulfate precipitation, and ion exchange chromatography Heat treatment, ammonium sulfate precipitation, ion exchange, and gel filtration chromatography
Lactobacillus reuteri
Bacillus circulans
(Expressed in Escherichia coli)
Streptococcus lactis
Bacillus licheniformis (cloned and expressed in Escherichia coli)
Bacillus megaterium
Bacillus stearothermophilus (expressed in Bacillus subtilis)
Sonication, ammonium sulfate precipitation, desaltation, column chromatography using DEAE–sepharose fast flow column, and affinity chromatography Ion exchange chromatography on Ni Sepharose 6 fast flow column and ultrafiltration
ZnCl2 precipitation, ion exchange membrane chromatography, hydrophobic interaction chromatography and gel filtration chromatography
Bacillus sp. 3088
Streptococcus thermophilus
Purified 292-fold by chromatography on Ultrogel ACA 34, DEAE–Sephadex A-50 columns and by affinity chromatography in agarose-p-aminophenyl- β-D-thiogalactoside
Purification procedure
Lactobacillus murinus
Source
Table 2. Different methodology for β-galactosidase purification.
1
Intracellular, M.W. is 530 kg mol 1. Optimum pH is 7.1. Galactose is competitive inhibitor (Ki 60 × 10 3 M), Km for ONPG and lactose are 0.98 × 10 3 and 6.9 × 10 3 M, respectively. Enzyme is most labile when it is suspended at cold (278.15 K) phosphate buffer. In presence of high concentration ammonium sulfate (0.85 M), enzyme is highly active and stabilizes. Intracellular, M.W. is 70 kg mol 1. Isoelectric point is 5.1. Optimum temperature and pH are 343.15 K and 7, respectively. Kinetics of thermal inactivation and half life at 338.15 and 343.15 K are 18 × 104 and 324 × 102 s. Km and Vmax are 2.96 × 10 3 M and 0.11 × 10 3 kmol s 1 kg of protein, respectively. Inhibitors are Fe(2+), Zn(2+), Cu(2+), Pb(2+), Sn (2+), and thiol agent. M.W. is 118 kg mol 1. Optimum pH 7.5–8.0 and temperature 328 K. Enzyme is stable at pH 6.0–9.0 and below 313 K. The Km and Vmax values for ONPG and lactose are 9.5 mM, 16.6 mM.min 1 and 12.6 mM, 54.4 mM. min 1, respectively. Homodimeric, optimum temperature, and pH are 323.15 K and 6.5, respectively. Km for lactose and ONPG are 169 × 10 3 and 13.7 × 10 3 M, respectively. Inhibitors are glucose and galactose. Metal activators are Na+, K+, Mg+2, Mn+2, and Ca+2.
Subunits are same molecular weight. M.W. is 170 kg mol . Maximum enzymatic activity at 318 K and pH 7 in presence of 50 mM phosphate buffer. The Km for ONPG and ONPG + 20 × 10 3 M of lactose are 480 × 10 6 and 870 × 10 6 M, respectively. Inhibitors are 10 × 10 3 M CaCl2, glutathione, and cysteine, and stimulator is 10 × 10 3 M MgCl2. Activators are mercaptoethanol and dithiotreitol. Enzymatic activity is not shown in presence of p-β-galactosidase. M.W. is 484 kg mol 1, associate with subunits 115, 86.5, 72.5, 45.7, and 41.2 kg mol 1. Optimum pH and temperature are 8 and 333.15 K, respectively. Isoelectric point is 6.2. Km is 6.34 × 10 3 and 6.18 × 10 3 M for ONPG and lactose, respectively. Inhibitors are galactose divalent Hg, Cu, and Ag. Metal activator is divalent Mg. EDTA do not affect the enzyme activity. Heterodimer enzyme, M.W. is 105 kg mol 1 (monomers are 72 and 35 kg mol 1). Isoelectric points are 4.6–4.8 and 3.8–4.0 for L. reuteri L103 and L. reuteri L461, respectively. Optimum pH is 6–8. Inhibitor is D-glucose and activators are Na (+), Mn (+2), and K (+). M.W. is 212 kg mol 1, composed by 145 and 86 kg mol 1. Km for three subunits are 3.6 × 10 3, 5.0 × 10 3, and 3.3 × 10 3 M for ONPG, respectively and those are 3.7 × 10 3, 2.94 × 10 3, and 2.71 × 10 3 M, respectively considering lactose as a substrate. Potential of GOS synthesis
Characteristics of purified enzyme
(Continues)
[55]
[54]
[53]
[52]
[51]
[50]
[49]
[48]
[47]
[46]
Reference
Asia-Pacific Journal of Chemical Engineering β-GALACTOSIDASE: A REVIEW
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
335
Ultrasonic treatment in the presence of Triton X-100
Ammonium sulfate precipitation, dialysis, anion exchange column (Mono Q HR 5/5, Pharmacia) using a FPLC system (Pharmacia). Active β-galactosidase fractions has been collected from several chromatographic runs and concentrated using the Centriplus system (Millipore). The active fraction of βgalactosidase has been obtained from ion exchange chromatography.
Column chromatographies by DEAE–Sephadex A-50,TSK gel Toyo Pearl HW-55S, and TSK gel DEAE-5PW
Bifidobacterium infantis (expressed in Escherichia coli)
Cryptococcus laurentii OKN-4
Ion exchange chromatography on DEAE– sepharose, affinity chromatography on PABTG-sepharose, and gel filtration on Sephacryl S-300 Ultrasonication and subsequently purified by Q Fast-Flow chromatography and gel chromatography on a Superose 6 HR column
Purification procedure
Bifidobacterium bifidum
Bifidobacterium longum CCRC 15708
Arthrobacter sp.
Source
Table 2. (Continued)
[56]
Intracellular, homodimeric, each subunit M.W. is 116 kg mol 1. Optimum pH and temperature are 6–8 and 298.15 K, respectively. Activators are thiol compounds, Na (+), and K (+). Inactivated by 4-chloromercuribenzoic acid, Pb (2+), Zn (2+), and Cu (2+) Finally 15.7-fold, a yield of 29.3%, and a specific activity of 168.6*106 U kg-1 protein (β-galactosidase) has been achhived. The MW was 357 kg.mol-1 as determined from Native-PAGE. The pH and temperature optima of the purified β-galactosidase were 7.0 and 323 K, respectively considering ONPG as a substrate,. The enzyme was stable at a temperature up to 313 K and at pH values of 6.5-7.0. Values of K m and V max for this purified enzyme were found to be 0.85 mM and 70.67 *106 U kg-1, respectively. It was found that Na+ and K+ stimulated the enzyme up to 10-fold, while Fe3+, Fe2+, Co2+, Cu2+, Ca2+, Zn2+, Mn2+ and Mg2+ inhibited the activity of βgalactosidase. Furthermore it was found that glucose, galactose, maltose, or raffinose exerted little or no effect on the β-galactosidase activity, lactose and fructose inhibited the enzyme activity. The effect of lactose on the enzyme activity for ONPG is probably a case of competitive inhibition. Enzyme has different subunits, such as 163, 170, 178, and 190 kg mol 1. The M.W. is 362 kg mol 1 and isoelectric point is 5.25. The purified enzyme is stable at temperatures below 318 K and over the pH ranges from 6.5–8. Lactose hydrolysis by the purified enzyme take places at pH 6.5 and temperature 310 K. The enzyme is tetramer, with an M.W. of about 470 kg mol 1, and its subunit is 115 kg mol 1. The optimum temperature and pH for ONPG and lactose are 333 K, pH 7.5 and 323 K, pH 7.5, respectively. The enzyme is stable over a pH range of 5.0–8.5 and remains active for more than 4800 s at pH 7.0, 323 K. The enzyme activity has been significantly increased by reducing agents. Maximum activity has been shown in presence of both Na+ and K+ at a concentration of 10 × 10 3 M. The enzyme is strongly inhibited by p-chloromercuribenzoic acid, divalent metal cations and Cr3+, and to a lesser extent by EDTA and urea. The hydrolytic activity using lactose as a substrate has been significantly inhibited by galactose. The Km and Vmax values for ONPG and lactose are 2.6 × 10 3 M, 262 × 106 U kg 1 and 73.8 × 10 3 M, 1.28 × 106 U kg 1, respectively. The enzyme possesses strong transgalactosylation activity. The production rate of GOS from 20% lactose at 303 and 333 K is 120 × 10 3 kg L 1, and this rate increases to 190 × 10 3 kg L 1 when 30% lactose has been used as a substrate. M.W. is 200 kg mol 1. homodimeric, optimum pH is 4.3, and enzyme is stable at pH between 2.8 and 9.3. The optimum temperature is 333 K and enzyme is stable at temperature below 330.5 K for 600 s
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
(Continues)
A. NATH ET AL.
[60]
[59]
[58]
[57]
Reference
Characteristics of purified enzyme
336 Asia-Pacific Journal of Chemical Engineering
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Precipitation with ammonium sulfate, ion exchange chromatography on DEAE–Sephadex, affinity chromatography, and chromate focusing Gel filtration chromatography, ion exchange chromatography, and affinity chromatography Gel filtration on Superose 12 PC 3.2/30 column and ion exchange
Kluyveromyces lactis
Kluyveromyces lactis
Protein precipitation by Pectinex Ultra SP followed by desalting and column chromatography using DEAE–Sepharose fast flow.
Aspergillus aculeatus
Penicillium chrysogenum
Gel filtration, anion exchange chromatography on DEAE–Sepharose CL-6B, hydrophobic chromatography on octyl-SepharoseCL-4B and cation exchange chromatography on CMSepharose CL-6B
Metal-ion affinity chromatography (IMAC) followed by size-exclusion separation
2-propanol fractionation, column chromatography on DEAE–Sephadex A-50, and Sephadex G-200
Purification procedure
Aspergillus niger
Aspergillus oryzae
Source
Table 2. (Continued)
incubation. The Km values of the enzyme are 18.2 × 10 3 M and 11.4 × 10 3 M, and the values of Vmax are 1.28 × 10 3 kmol s 1 kg of protein, and 0.09 × 10 3 kmol s 1 kg of protein for ONPG and lactose, respectively. The enzyme is strongly inhibited by Hg2+, Ag+, 2-mercaptoethanol, glucose, maltose, and maltotriose. Enzyme is potential of GOS synthesis. Extracellular, optimum pH for ONPG and lactose are 4.5 and 4.8, respectively. Optimum temperature is 319.15 K. Km are 7.2 × 10 4 and 1.8 × 10 2 M for ONPG and lactose respectively. Inhibitors are divalent Hg, Cu, N-bromosuccinimide, and sodium laurylsulfate. Apparent M.W. is 105 kg mol 1. Extracellular, M.W. is 113 kg mol 1. Mutant enzyme has five times higher catalytic activity on the synthetic substrate ONPG compared with the wild-type enzyme. Moreover, the mutant enzyme is more thermo resistant compared with the wild type. Glycoprotein in nature. Associates with three subunits, and each M. W. are 124, 150, and 173 kg mol 1. Isoelectric point is 4.6. Optimum pH lies between in 2.5 to 4.0. Heat stable up to 333.15 K. Km value varies from 85 × 10 3 to 125 × 10 3 M for lactose. Km value varies 2.4 × 10 3 M for ONPG. Vmax values are 104 × 103 unit enzyme kg of protein and 121 × 103 unit enzyme kg of protein at 303.15 K for ONPG and lactose, respectively. The M.W. is 120 kg mol 1 (approximately), isoelectric point lies between 5.3 and 5.7 and is optimally active at pH 5.4 and temperature 328–333 K. Based on the N-terminal amino acid sequence, the enzyme probably belongs to family 35 of the glycosyl hydrolases. Enzyme is potential of synthesizing GOS. Enzyme has activity towards two exo-poly-saccharide (EPSs), having lactosyl side chains attached to different backbone structures. The enzyme degraded O-deacetylated EPS B891 faster than EPS B39. Moreover, the presence of acetyl groups in EPS B891 slows down the hydrolyzing rate but the enzyme is still able to release all terminally linked galactose. Intracellular, Specific activity is 5.84 × 103 U kg of protein. Optimum temperature and pH are 303.15 K and 4, respectively. Km and isoelectric point are 1.81 × 10 3 M and 4.6 for ONPG. Multimeric enzyme, M.W. is 270 kg mol 1, and M.W. of single monomer is 66 kg mol 1 Affinity chromatography is shown better results than gel permeation chromatography, and ion exchange chromatography. Similar molecular weight subunit. Enzyme is confirmed by specific antigenantibody reaction (ELISA test). Intracellular, M.W. of single monomer is 124 kg mol 1
Characteristics of purified enzyme
(Continues)
[66]
[65]
[41]
[64]
[63]
[62]
[61]
Reference
Asia-Pacific Journal of Chemical Engineering β-GALACTOSIDASE: A REVIEW
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
337
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Gel filtration chromatography (Ultrogel AcA 34), anion exchange (Mono Q), and gel filtration (Superose-12)
Centrifugation, cell disruption, heat treatment, DEAE–Toyopearl chromatography, salting out, Butyl-Toyopearl chromatography, Chromatofocusing chromatography, and PATG chromatography
Ammonium sulfate fractionation, column chromatography on Sephadex G-100, and DEAE–Sephadex A-50, 78-fold purification FPDA 13 column chromatography, ammonium sulfate fractionation, Ultrogel AcA34 column chromatography, DEAE–Sepharose CL-6B column chromatography, TSK gel DEAEToyopearlpak 650S High-pressure liquid chromatography (HPLC)
chromatography on Mono Q PC 1.6/5 column using a FPLC system Cell disruption, DEAE–Sephadex ion exchange chromatography, and chromatography on hydroxylapatite Extraction with 2% chloroform, acetone, and ammonium sulfate precipitation
Purification procedure
Extracellular thermostable, relative M.W. of 145 000 and s°20,w of 7.1 s. Michaelis constant Km is 0.75 × 10 6 kM and molecular activity (kcat) is 63.1 s 1 at pH 7.2 and 328 K for ONPG where as Km is 0.04 × 10 6 kM and kcat is 55.8 s 1 for p-NPG. Enzyme has a high transgalactosylation activity. The enzyme reacts with 1.75 M lactose at 333 K and pH 7.0 for 7.92 × 104 s to obtain maximum yield oligosaccharides (41% (w/w)). The general structure for the major transgalactosylic products can be expressed as (Gal)cGlc, where n is 1, 2, 3, and 4 with a glucose at a reducing terminal. M.W. is 170 and 86 kg mol 1 for denatured enzyme, isoelectric point 4.1. The optimal temperature is 358 K, and it is stable at temperatures up to 353 K for 3 600 s. The optimal pH range for the enzyme is 4.5 to 5.0 and the enzyme is stable at pH 2.5 to 7.0. Ezyme is inhibited by Hg2+. The Km values for ONPG and l actose are 9.5 × 10 3 and 2.4 × 10 3 M, respectively. The Vmax for these substrates are 96 mol s 1 kg of protein and 4 mol s 1 kg of protein, respectively. The enzyme possesses a high level of transgalactosylation activity (higher yield 39% from 200 g L 1 lactose), which produces GOS including tri- and tetrasaccharides. M.W. is 700 kg mol 1 and subunit is 59 ± 1 kg mol 1. Isoelectric point is 4.9. The optimum temperature and pH for enzyme activity are 353 K and 5.5, respectively. The enzyme is stable over a wide pH range (pH 3–12), and the thermostability of the enzyme is enhanced by CaCl2. The enzyme is significantly activated by alkali and alkaline-earth-metal salts. Inhibitors are thiol-binding agents, glucose, and galactose. The enzyme specific for b-D anomeric linkages and the identity of the aglycone moiety also influenced enzyme activity dramatically. Enzyme is potential of GOS synthesis.
M.W. is 135 kg mol 1. Optimum pH is 7.25. Km is 12 × 10 3 to 17 × 10 3 M for lactose. Km is 1.6 × 10 3 M for ONPG. Metal activators are Na(+) and Mn(+2). M.W. is 280 kg mol 1. Optimum temperature and pH are 318.15 to 325.15 K and 6.2, respectively. Inactivates in 240 s at 329.15 K. Km are 3.1 × 10 3 and 25 × 10 3 M for ONPG. Inhibition constants are 58 × 10 3, 110 × 10 3, 111 × 10 3, and 52 × 10 3 M for ONPG, galactose, ribose, and lactose. Inhibitors are also P-chloromercuribenzoate and dithiothreitol. Optimum pH is 7.2. Metal activator divalent magnesium, Km is 1.18 × 10 3 M for ONPG.
Characteristics of purified enzyme
[72]
[71]
[70]
[69]
[68]
[67]
Reference
A. NATH ET AL.
Thermus aquaticus
Sterigmatomyces elviae
Saccharopolyspora rectivirgula
Saccharomyces lactis
Kluyveromyces marxianus
Source
Table 2. (Continued)
338 Asia-Pacific Journal of Chemical Engineering
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering
CHARACTERIZATION OF β-GALACTOSIDASE Apart from catalyzing the hydrolysis of β-galactosides into monosaccharides by lactase, the enzyme may also cleave fucosides and arabinosides with much lower efficiency. Lactase is often confused as an alternative name for β-galactosidase, but it is merely a sub-class of β-galactosidase. In other words, β-galactosidase is an exoglycosidase, which hydrolyzes the β-glycosidic bond formed between galactose and its organic moiety.[73] Molecular weight, amino acids chain length, position of the active site, pH, and optimum thermal stability are significantly differed by the microbial sources.[74] The choice of suitable β-galactosidase source depends on the condition of reaction. For example, dairy yeasts with optimum pH (6.5–7.0) are habitually used for the hydrolysis of lactose in milk or sweet whey.[75] On the other hand, the fungal β-galactosidases with optimum pH (3.0–5.0) are more suitable for acidic whey hydrolysis.[76] The activity of different β-galactosidases also depends on presence of ions. The fungal β-galactosidases are active without presence of ions as cofactors; where β-galactosidase isolated from K. lactis shows its higher activity in presence of Mn2+ and Na+. β-galactosidase synthesized from K. fragilis are mostly active in presence of Mn 2+ , Mg 2+ , and K + . [77] On the contrary, Ca 2+ and heavy metals inhibit the enzyme activity of all β-galactosidases.[78] Properties of β-galactosidase synthesized by different microorganisms are described in Table 3. In 1970, 1024 amino acids of β-galactosidase of E. coli were first sequenced.[93] After 24 years, four chains comprising the protein were discovered to be 464 kg mol 1 tetramer with 222-point symmetry. Every unit of β-galactosidase contains five domains; whereas the active site persists in the third domain. This enzyme can be split into two peptides, LacZα and LacZΩ, none of them is active but both spontaneously reassemble a functional enzyme. This characteristic is used for many cloning vectors to achieve α-complementation in specific laboratory strains of E. coli, where the plasmid encodes the small LacZα while the large LacZΩ is encoded by the bacterial chromosome. Aftermath, when DNA fragments inserted in the vector, production of LacZα disrupted, the cells revealed no β-galactosidase activity, were subjected to the blue/white screening of recombinant clones further.[94] The active site of β-galactosidase catalyzes the hydrolysis of disaccharide substrate via ‘shallow’ and ‘deep’ binding. The beta-linkage of the substrate was cleaved by a terminal carboxyl group on the side chain of glutamic acid.[95] It has been determined that the DNA fragment of thermophilic bacterium Thermoanaerobacter ethanolicus contains three open reading frames. One of the open reading frames corresponded to the LacA gene for a thermostable s-galactosidase and the native recombinant LacA showed the highest activity at © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
β-GALACTOSIDASE: A REVIEW
348 K–353 K. Immobilized on aldehyde silochrome, LacA was even more thermo stable and retained high activity.[96] It has been found that individual molecules of β-galactosidase from the crystallize enzyme as well as the original enzyme displayed a range of activity of 20-fold or greater. Molecules obtained from two diverse crystals have identical activity distributions, i.e., 31 600 ± 1100 and 31 800 ± 1100 reactions per minute per enzyme molecule. This activity of the enzyme was found to be drastically different from that of the enzyme, which was used to grow the crystals (showed an activity distribution of 38 500 ± 900 reactions per minute per enzyme molecule).[97] Induced β-galactosidase in E. coli wild-type strains ATCC 8677 and 35321 in the presence of various protease inhibitors have been studied. The presence of the protease inhibitors had least effect on the average distribution of single molecule activities, and the relative activities of the enzyme for the diverse substrates differed between the strains.[98] Averagecombined turnover numbers of the enzyme from wildtype E. coli strains ATCC 35321 and 8677 in vivo and in vitro conditions in the presence and absence of His6 tag differed considerably. This indicated that synthesized enzymes in both conditions (vivo and vitro) were not alike and presence of a C-terminal His6 tag altered the activity of s-galactosidase.[99] Moreover, it was found that electrophoretic mobility and catalytic activity of individual molecules of β-galactosidase synthesized by E. coli were different, although they had potentiality to act on the same substrate molecule.[100]
IMMOBILIZATION OF β-GALACTOSIDASE Immobilization has shown to improve the stability of β-galactosidase, reusages, and reduces the processing time in food and other industries. For example, the immobilization of β-galactosidase of Thermus sp. T2 was performed using ionic adsorption by a new anionic exchanger resins (based on coating of Sepabeads internal surfaces with polyethylenimine) and conventional DEAE–agarose. Immobilization was carried out in both cases, but the adsorption strength showed much greater in the case of PEI–Sepabeads than in DEAE supported at both pH 5.0 and 7.0. Also, the PEI–Sepabeads remained wholly active, and after several weeks of incubation at 323 K, it showed the lactose hydrolysis in milk.[101] Also a new hetero functional epoxy supports were used for immobilization of β-galactosidase. The capability of a standard Sepabeads-epoxy supports to immobilize β-galactosidase from Thermus sp. strain T2 that equalized with other Sepabeads-epoxy, which supports partially modification using boronate, iminodiacetic, metal chelates and ethylenediamine. Here, immobilization was depended on the support, ranging from 95% to 5% using Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
339
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Archae
Fungus
Arthrobacter psychrolactophilus Bacillus circulans Bacillus megaterium Bacillus stearothermophilus Bifidobacterium adolescentis Bifidobacterium bifidum Bifidobacterium infantis Cryptococcus laurentii Enterobacter agglomerans Lactobacillus acidophilus Lactobacillus reuteri Streptococcus pneumonia Alicyclobacillus acidocaldarius Caldicellulosiruptor saccharolyticus Geobacillus stearothermophilus Saccharopolyspora rectivirgula Thermotoga maritima Thurmus sp. Thurmus aquaticus Kluyveromyces fragilis Kluyveromyces lactis Sporobolomyces singularis Sterigmatomyces elviae Bullera singularis Aspergillus oryzae Aspergillus aculeatus Sulfolobus solfataricus
Name of consortium 8 5–6 6–9 7 6 6.5 7.5 4.3 7.5–8 6.5–8 6–8 5.5–7.5 5.5 6 6.5 6.5–7.2 6.5 6.5 5.5 6.5 6.6–7 4 4.5–5 5 4.8 5.4 6.5
338 333 353–358 343 353 303 310–313 303 358 323 303 328–333 348
pH
333 323 313 343 323 310 323–333 331 310–313 328 323 303 338 353
Temperature (K)
20 5.1 60 125 526 — 569 12 — 230 180 — 592 211 0.5 — 70 — 5.7 — — 8.7 20 56 40 24 116
— 1.44 × 104 (333 K) 960 (363 K) 7.2 × 104 (343 K) — 1.8 × 104 (313 K) — — 3600 (353 K) 7200 (318 K) 600 (343 K) — 1.08 × 104 (358 K)
Specific activity (U mg 1)
3600 (303 K) — — 3.24 × 104 (343 K) 600 (323 K) 1.44 × 104 (318 K) 7200 (333 K) 600 (331 K) — 1.728 × 105 (310 K) — — 360 (353 K) 1.728 × 105 (343 K)
Half life (s)
[92]
[62] [64]
[90] [71] [91]
[88] [89]
[87] [88] [72]
[86] [70]
[84] [85]
[82] [48] [83]
[60] [81]
[80] [58] [59]
[54] [53]
[79] [50]
Reference
A. NATH ET AL.
Yeast
Bacteria
Type of consortium
Table 3. Properties of β-galactosidase synthesized by different microorganisms.
340 Asia-Pacific Journal of Chemical Engineering
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering
Sepabeads-epoxy-chelate, Sepabeads-epoxy-amino, or Sepabeads-epoxy-boronic using Sepabeads-epoxy-IDA. Amazingly, the immobilized β-galactosidase derivatives showed outstandingly good result but different stabilities had been notified after favoring multipoint covalent attachment by long-term alkaline incubation. The enzyme immobilized on Sepabeads-epoxy-boronic was found to be the steadiest. The crosslinking with aldehyde-dextran allowed the stabilization of the quaternary structure of the enzyme. The optimal derivative was extremely active in lactose hydrolysis even at 343 K (over 1000 IU g 1), maintaining its activity after extended incubation times with no risk of product contamination with enzyme subunits.[102] Protocol for immobilization of β-galactosidase synthesized by E. coli using diverse supports (glyoxyl, epoxy, BrCN groups, or by glutaraldehyde crosslinking on matrix, containing primary amino groups), and strategies have been studied. In each case, the immobilization yield showed 100% with active recoveries between 50% and 100% (using ortho-nitrophenylβ-galactoside as substrate). Ratio of synthetic activity to hydrolytic activity (Vs/Vh) was lower than 0.1 when soluble enzyme and the Eupergit 250 L enzyme were immobilized on BrCN at 277 K and pH 7.0, resulting 0.46 and 0.8, respectively.[103] Immobilization of β-galactosidase producing permeabilized dead cells of K. lactis ATCC 8583 into gelatin using glutaraldehyde as crosslinker was performed, where 30% activity obtained by immobilized cells relative to free disrupted cells.[104] The usage of calcium alginate, Κ-carrageenan, and gellan-xanthan gel beads for entrapment of β-galactosidase synthesized by Streptococcus thermophilus, enhanced the stability of enzyme at higher temperatures (>298 K).[105] Solid state fermentation with co-immobilized β-galactosidases synthesized by K. lactis, Aspergillus oryzae, and yeasts in polyvinyl alcohol hydrogel lens-shaped capsules have been performed. In the process the enzyme, synthesized from Kluyveromyces lactis and Saccharomyces cerevisiae showed the highest activity (galactose output increased from 3 to 4.1 g l 1 h 1), which reduced the Different reduction of processing time.[106] methodologies for immobilization of β-galactosidases have been described in Table 4.
INDUSTRIALIZED APPLICATIONS OF β-GALACTOSIDASE Microbial β-galactosidase plays a tremendously essential role in the production of various industrial relevant products such as biosensor, lactosehydrolyzed milk, ethanol, and GOS, also it has been used in the field of bioremediation, diagnosis, and in treatment of lactose digestion disorder etc. © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
β-GALACTOSIDASE: A REVIEW
Use of β-galactosidase in biosensor A biosensor associated with two distinct enzymatic activities (β-galactosidase and glucose peroxidase) has been developed for quantitative detection of lactose in commercial samples of milk. To avoid interferences with glucose, a degree of different mode of measurement was done using biosensor.[146] Foresighting this technique, presumed β-galactosidase from Streptococcus mitis with a choline-binding domain was identified recently. This remarkable property makes it differentially functional property for biotechnological applications.[147] Intended as lactose-hydrolyzed milk production The cold–stable properties of Arthrobacter sp. 32c synthesized β-galactosidase could be useful for commercial, industrial conversion of lactose into galactose and glucose in milk products.[56] It has been reported that rudimentary β-galactosidase extract produced by Lactobacillus ssp. bulgaricus CHR Hansen Lb-12 was applied in sterile milk, which has been processed through ultra-high temperature method (UHT milk) for hydrolyzing lactose. Optimum amount of crude β-galactosidase extract and Maxilact enzyme for producing lactose-hydrolyzed milk was observed to be 0.418 and 0.512 U mL 1 respectively during 6 h of processing. Using more than 418 U L 1 of crude β-galactosidase extract showed undesirable acidity of lactose-hydrolyzed milk that significantly increased at temperature of between 288 and 290 K, while enrichment of acidity in lactose-hydrolyzed milk produced through Maxilact enzyme was not significant. Total count of lactose-hydrolyzed milk by 418 U L 1 of crude β-galactosidase extract, after 2.16 × 104 s of processing was significant high (5 to 30 Colony Forming Unit). Sensory estimation of lactose-hydrolyzed milk and ordinary ultra-high temperature milk (controlled) did not show any major differences with respect to acceptability of sweetness, taste, and color.[33] The difficulty in enzyme extraction and poor permeability of cell membrane to lactose was solved when permeabilized Kluyveromyces marxianus NCIM 3465 cells were used for the production of lactose-hydrolyzed milk. The ethanol-permeabilized yeast cells showed 89% of hydrolysed lactose under optimized conditions.[148] Role of β-galactosidase in whey utilization The execution of profound environmental legislation for recycling and reuse of waste materials is grabbing lots of headlines at present century. Since many years, dairy products have been used as valuable ingredients by the confectionery industry, being a huge economical source of capital, especially in the tropical and subtropical countries. The large volume of byproduct Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
341
342
Asia-Pacific Journal of Chemical Engineering
A. NATH ET AL.
Table 4. Different techniques for immobilization of β-galactosidases.
Immobilization method
Source of β-galactosidase
Covalent binding Covalent binding Entrapment Physical adsorption Physical adsorption Covalent binding
Aspergillus Aspergillus Aspergillus Aspergillus Aspergillus Aspergillus
Covalent binding Covalent binding Entrapment Physical adsorption Covalent binding Physical adsorption Covalent binding Entrapment Physical adsorption Physical adsorption
Aspergillus oryzae Aspergillus oryzae Aspergillus oryzae Aspergillus oryzae Aspergillus oryzae Aspergillus niger Kluyveromyces lactis Kluyveromyces bulgaricus Kluyveromyces fragilis Kluyveromyces fragilis and Kluyveromyces lactis Kluyveromyces fragilis Kluyveromyces fragilis Kluyveromyces lactis Kluyveromyces lactis Kluyveromyces lactis, Aspergillus oryzae and Saccharomyces cerevisiae Kluyveromyces lactis Kluyveromyces lactis Kluyveromyces lactis Kluyveromyces fragilis Thermus aquaticus YT-1 Thermus sp. T2 Penicillium expansum F3 Saccharomyces anamensis Pisum sativum Lactobacillus bulgaricus Bacillus stearothermophilus Bacillus circulans Bacillus circulans Escherichia coli Escherichia coli Escherichia coli Escherichia coli (Recombinant β-galactosidase) Escherichia coli Escherichia coli
Physical adsorption Physical adsorption Covalent binding Physical adsorption Entrapment Covalent binding Covalent binding Covalent binding Covalent binding Entrapment Physical adsorption Entrapment Covalent binding Physical adsorption Covalent binding Physical adsorption Physical adsorption Covalent binding Entrapment Covalent binding Physical adsorption Covalent binding Covalent binding Covalent binding
oryzae oryzae oryzae oryzae oryzae oryzae
of the dairy industry, namely, whey, creates severe disposal problem. Concerning about environmental aspects, research has been emphasized on membrane separation technology, providing remarkable new opportunities for large-scale protein and lactose fractionation. They intrude proteins that are multifunctional food ingredients of high nutritional value and offer a wide range of functional properties allowing development of new products and optimization of existing products within considerable low cost, but presence of high concentration of lactose in aqueous medium after separation of proteins from whey creates severe disposal problem.[149–153] Different strategies © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Immobilization matrix
References
Amino-epoxy Sepabead Chitosan bead and nylon membrane Nylon-6 and zeolite Phenol-formaldehyde resin Polyvinyl chloride and silica gel membrane Polyvinyl alcohol hydrogel and magnetic Fe3O4-chitosan as supporting agent Silica Silica gel activated with TiCl3 and FeCl3 Spongy polyvinyl alcohol Cryogel Celite and chitosan Cotton cloth and activated with tosyl chloride Porous ceramic monolith Magnetic polysiloxane-polyvinyl alcohol Alginate using BaCl3 Cellulose beads Chitosan
[107] [108] [109] [110] [111] [112] [113] [114] [115] [116] [117] [118] [119] [120] [121] [3]
Chitosan Chitosan bead Cotton fabric CPC-silica and agarose Poly(vinylalcohol) hydrogel
[122] [123] [124]
Corn grits Thiosulfinate/thiosulfonate Graphite surface Silica-alumina Agarose bead PEI–Sepabeads, DEAE–agarose Calcium alginate Calcium alginate Sephadex G-75 and chitosan beads Egg shells Chitosan Polyvinyl chloride and silica Eupergit C (Spherical acrylic polymer) Polyacrylamide gel Polyvinyl alcohol Chromosorb-W Cyanuric chloride-activated cellulose
[127]
Hen egg white Gelatine
[144]
[125] [126]
[128] [129] [130] [131] [132] [133] [134] [135] [136] [137] [138] [139] [140] [141] [142] [143]
[145]
have been attempted for whey lactose conversion, those are related to the synthesis of nutraceuticals GOS, as well as alcohol from whey permeates would ensure ultimate consumption and utilization of casein whey.[154,155] Function of β-galactosidase in ethanol fermentation The idea of kinetic analysis of alcoholic fermentation of lactose using a recombinant flocculent strain of Saccharomyces cerevisiae NCYC869-A3/T1, expressing both the LAC4 (coding for β-galactosidase) and LAC12 (lactose permease) genes of Kluyveromyces Lactis conceded. The lactose was Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering
completely utilized in all fermentative processes, and it has been observed that with increase of initial lactose concentration (5 to 200 g L 1), the level of ethanol production increased linearly.[156] Meticulously, kinetic model with respect to biomass growth, lactose hydrolysis, and ethanol production using β-galactosidase synthesized by Kluyveromyces sp. was performed considering whey permeate and cheese whey powder as growth medium.[157–159] Moreover, Saccharomyces cerevisiae has also been reported for production of ethanol considering concentrated deproteinized whey, cheese whey powder, and salted cheese whey as feed stock.[160–162] Ethanol fermentation of cheese whey powder solution using the pure culture of Kluyveromyces marxianus (DSMZ 7239) was studied in packed column bioreactor using olive pits as sustaining particles for cell attachment.[163] Researches have also been conducted for ethanol production by membrane recycle bioreactor and using co-immobilized S. cerevisiae strain and β-galactosidase in semicontinuous fermentation process considering whey permeate and whey medium, respectively.[164,165] Thus, whey utilization by β-galactosidase reduces the burden of water pollution and establishes the concept of recycling and reuse of waste materials. Galato-oligosaccharide production by β-galactosidase In recent years, many investigations have been carried out in the field of prebiotics, considered as functional food. Among them, one of the recognized functional food ingredients is oligosaccharides. They are
β-GALACTOSIDASE: A REVIEW
carbohydrates containing three to ten sugar units bound with glycosidic bonds. It has been seen that there are a number of classes of oligosaccharides, but among them, GOS has attracted particular attention because of their presence in human breast milk. GOS are nondigestible, carbohydrate-based food ingredient, responsible for human and animal nutrition. Production of GOS by transgalactosylation activity results the formation of 4′- or 6′-galactosylactose, longer oligosaccharides, transgalactosylated disaccharides, and nonreducing oligosaccharides in presence of β-galactosidases. Depending on the source of enzyme and conditions of reaction, various glycosidic linkages, such as β(1,2), β(1,3), β (1,4), and β (1,6), are formed in the end product (GOS).[166] It has been found that the amount of GOS synthesis from lactose depends on the initial concentrations of lactose present in the reaction mixture instead of the concentration of β-galactosidase. There are different phenomena that have been reported to elucidate the synthesis mechanism of GOS, which are depicted in Figs 4–6. It is believed that the presence of GOS in human milk influences the growth of bifidobacteria in the gastrointestinal tract of newly born and breast-fed infants. GOS fraction (referred to as bifidus factor) in cow’s milk also provides several health benefits.[170] GOS is stable under acidic conditions during food processing as well as maintain excellent taste quality, which makes them popular as an active ingredient in a wide variety of food products. They pass through the small intestine without being digested for low caloric value. In addition, GOS is not metabolized by
Figure 4. Schematic diagram of galato-oligosaccharide synthesis from lactose,
(A) engineering approach and (B) biochemical approach (Figure adapted from Boon et al. (1999)).[167] © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
343
344
A. NATH ET AL.
Figure 5. Schematic diagram of galato-oligosaccharide synthesis from lactose (Figure adapted from Neri et al. (2008)).[168]
Figure 6. Schematic diagram of galato-oligosaccharide synthesis from lactose (Figure adapted from Palai et al. (2012)).[169]
microorganisms in the oral cavity.[171] The detail works carried out in this direction have been represented in Table 5. Applications β-galactosidase in medical and immunology research Clostridium perfringens ATCC 10543 synthesized endo β-galactosidase, which is capable of liberating beneficial A trisaccharide and B trisaccharide from glycoconjugates containing blood group A and B glycotopes, respectively, was isolated by Anderson et al., 2005. Recombinant EABase damages the blood group A and B antigenicity of human type A and B erythrocytes with the release of A-Tri and B-Tri from blood group A + and B + containing glycoconjugates. Here, the incomparable specificity of β-galactosidase was useful for studying the structure and function of blood group A-containing and B-containing glycoconjugates.[193] The recombinant endo-β-galactosidase (ABase), which releases A/B antigen was developed in 2009. It removed 82% of A antigen and 95% of B antigen from human A/B red blood cells and concealed anti-A/B antibody binding, also the complement activation effectively. In vivo infusion into a blood type A demonstrated the reduction of A antigen expression in the glomeruli of kidney (85% at 3600 s, 9% at 1.44 × 104 s, and 13% at 8.64 × 104 s) and the sinusoids of liver (47% at 3600 s, 1% at 1.44 × 104 s, and 3% at 8.64 × 104 s) without any grave adverse effects. This substitute approach remains useful for minimizing © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Asia-Pacific Journal of Chemical Engineering
antibody removal and anti-B cell immunosuppression as an adjuvant therapy of ABO incompatible kidney, liver, and possibly heart transplantation.[194] Expression of β-galactosidase, synthesized by E. coli within muscle fibers has been demonstrated by Liu and Roffler, 2006. They conclude that repeated intramuscular injections of β-galactosidase could encourage strong immune responses among immuno-competent animals and cause elimination of transduce muscle fibers by inflammatory cells.[195] Recently, β-galactosidase from the mesoacidophilic fungus Bispora sp. MEY-1 under simulated gastric conditions has shown greater stability (100%) and hydrolysis ratio (>80%) toward milk lactose than commercially available β-galactosidase from Aspergillus oryzae ATCC 20423. Thus, this β-galactosidase may be a better digestive supplement for alleviating symptoms associated with lactase deficiency.[196] Recombinant β-galactosidases in cooperating one or two different peptides from the foot-and-mouth disease virus (FMDV) nonstructural protein 3B per enzyme monomer, granted specified differentiation between sera of FMDV-infected pigs, cattle, and sheep, and those of native and conventionally vaccinated animals. These FMDV infection-specific biosensors can provide effectual and versatile alternatives for the serological differentiation of FMDV-infected animals.[197]
SCOPE FOR FURTHER STUDY β-galactosidase belongs to lactase group, which hydrolyzes β-glycosidic bond formed between galactose and its organic moiety. Moreover, in some cases, it takes part in transgalactosidase reaction. The disadvantage of wild-type β-galactosidase is its lower activity as well as it is inhibited by hydrolyzed product, glucose, and galactose. Therefore, research regarding transgenic β-galactosidase synthesis will boost up hydrolytic activity as well as GOS synthesis. Although, synthesis mechanism of β-galactosidase is well established for E. coli, still much more information is needed for other microorganism. Although, it is recommended that probiotic consortium (lactic acid bacteria) is much more acceptable for food grade β-galactosidase synthesis, but because of product inhibition by lactic acid, research on bioprocess design is in demand. More than 90-fold purification of β-galactosidase is yet to achieved. In this circumstances, suitable purification device as well as development of purification protocol for high throughput is required. Research in the line of process intensification of industrial β-galactosidase production and purification should be a great challenge. It is obvious, immobilized β-galactosidase provides several benefits for GOS synthesis but due to lower activity at immobilize condition; studies on the active site of enzyme and Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering
β-GALACTOSIDASE: A REVIEW
Table 5. Different microorganisms for galato-oligosaccharide synthesis and parameters of transgalactosylation reaction.
Nature of enzyme Crude Crude Crude Crude Recombinant Crude Crude Purified Purified Purified Purified Recombinant Recombinant Recombinant Whole cells Toluene-treated cells Immobilized Immobilized Immobilized Immobilized Immobilized cells Immobilized cell Crude Crude Purified Purified Recombinant Recombinant Immobilized Immobilized Immobilized Immobilized cell
Temperature (K)
Enzyme source Aspergillus oryzae Bacillus circulans Bifidobacterium longum Geobacillus stearothermophilus Geobacillus stearothermophilus R109W Lactobacillus reuteri Talaromyces thermophilus Bullera singularis Enterobacter agglomerans Lactobacillus acidophilus Penicillium simplicissimum Bifidobacterium infantis Sulfolobus solfataricus Thermotoga maritima Bifidobacterium bifidum Rhodotorula minuta Aspergillus candidus Bacillus circulans Bullera singularis Kluyveromyces lactis Talaromyces thermophilus Cryptococcus laurentii Talaromyces thermophilus Penicillium sp. Saccharopolyspora rectivirgula Sterigmatomyces elviae Pyrococcus furiosus Thermus sp. Sirobasidium magnum Sirobasidium magnum Aspergillus oryzae Sporobolomyces singularis
choice of suitable matrix are required. Without any contradiction, β-galactosidase plays several roles in biopharmaceutical, food, and dairy industries. Research for more applications in several fields particularly cancer detection and pharmaceutical research will boost up immunological research and biotechnological economy.
CONCLUSION In this review, authors have tried their best to accumulate all the information regarding production, purification, characterization, immobilization, and application of β-galactosidase. β-galactosidase is one of the important enzyme that not only offers nutritional applications but also used for waste treatment. β-galactosidase used in food, pharmaceuticals, and dairy industries for producing low lactose-containing food product, sweeteners, GOS, and also used for dairy waste treatment. It is also used for ethanol production, which is a challenging step in present century. Several microbes, especially probiotic consortia are recommended for enzyme production using casein whey as a growth medium. Purified β-galactosidase will ensure © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
313 313 318 310 310 303 313 323 323 303 323 333 353 353 312 333 313 313 318 313 313 313 313 328 333 333 353 343 323 323 313 328
pH 4.5 6 6.8 6.5 6.5 6.5 6.5 5 7.5 6.5 6.5 7.5 6 6 6.8 6 6.5 6 3.7 7 6.5 4.3 6.5 4 6.5 5 5 7 — — 4.5 5.0
Synthesis rate (g L 1 h 1) 24 2.2 13 0.4 6.9 3.9 13 3.9 3.9 7.9 11 13 5.6 18 65 3.2 87 4.2 4.8 25 18.75 3.6 — — — 3.2 — — 3.2 5.8 106 8.7
Reference [172] [173] [174] [86] [86] [175] [176] [91] [81] [82] [177] [59] [178] [179] [180] [181] [182] [183] [184] [185] [186] [187] [186] [188] [70] [71] [189] [190] [191] [191] [117] [192]
the high activity as well as poses better catalysis properties. Enzyme immobilization provides enzyme reutilization, also the immobilized enzyme showed high level of hydrolysis and thus it can be applied successfully for hydrolyzing lactose in milk and whey. The isolation of pyschrophilic bacteria with cold active β-galactosidase has opened up the possibility of processing of GOS at low temperatures. On the other hand, thermostable enzyme has the unique ability to retain their activity at higher temperature for prolonged periods, and the process is less prone to microbial contamination due to higher operating temperature. Therefore, it may be concluded that utilization of β-galactosidase is a twofold solution of biotechnology economy and waste treatment. Furthermore, this work will provide a ready reference for future scientific research regarding β-galactosidase.
Acknowledgements Arijit Nath acknowledges Council of Scientific and Industrial Research (CSIR), New Delhi for financial Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
345
346
Asia-Pacific Journal of Chemical Engineering
A. NATH ET AL.
support as an SRF. The reported work is a part of a University Grants Commission (UGC) Major project, entitled ‘Production and Purification of βgalactosidase from Milk Whey-based Lactic Acid Bacteria using Fermentation and Membrane-based Separation Techniques”. The contribution of UGC is gratefully acknowledged.
REFERENCES [1] M.L. Shuler, F. Kargi. Bioprocess Engineering Basic Concept, 2nd edn, Prentice-Hall Inc, New Jersey, 2002. [2] M.L. Sanz, G.R. Gibson, R.A. Rastall. J. Agri. Food Chem., 2005; 53(13), 5192–5199. [3] C.E. Rycroft, M.R. Jones, G.R. Gibson, R.A. Rastall. J. Appl. Microbiol., 2001; 91(5), 878–887. [4] M.C.E. Lomer, G.C. Parkes, J.D. Sanderson. Aliment. Pharm. Ther., 2008; 27, 93–103. [5] T. Sahi. Scand. J. Gastroenterol., 1994; 29(202), 7–20. [6] J. Jussila, M. Isokoski, K. Launiala. Scan. J. Gastroenterol., 1970; 5, 49–56. [7] Z. Mlichová, M. Rosenberg. J. Food Nutr. Res., 2006; 45(2), 47–54. [8] S.P.S. Panesar, S. Kumari, R. Panesar. Enzyme Res., 2010, 1–16. [9] F. Jacob, J. Monod. J. Mol. Biol., 1961; 3, 318–356. [10] K.L. Anderson, G. Purdom. In Proceedings of the Sixth International Conference on Creationism Creation Science Fellowship and Dallas, Texas, (Ed.: A.A. Snelling), Institute for Creation Research, Pittsburgh, Pennsylvania, 2008; pp. 149–163. [11] B. Alberts, A. Johnson, J. Lewis, M. Raff, K. Roberts, P. Walter. Molecular Biology of the Cell, 5th edn, Garland Science, New York, 2007; pp. 1392. ISBN:9780815341055. [12] http://academic.pgcc.edu/~kroberts/Lecture/Chapter%207/ regulation.html. Accessed date 18th june, 2013 [13] B. Görke, J. Stülke. Nat. Rev. Microbiol., 2008; 6(8), 613–624. [14] A.J.F. Griffiths, J.H. Miller, D.T. Suzuki, Lewontin R.C., W.M. Gelbart. An Introduction to Genetic Analysis. 7th edition, New York, 2000; ISBN-10: 0-7167-3520-2 [15] L.H. Hansen, S. Knudsen, S.J. Sønrense. Curr. Microbiol., 1998; 36(6), 341–347. [16] S.A.-A. Ismail, Y. EI-Mohamady, W.A. Helmy, R. Abou-Romia, A.M. Hashem. Aust. J. Basic Appl. Sci., 2010; 4(10), 5051–5058. [17] N.P. Laxmi, M.A. Mutamed, P.S. Nagendra. Int. Food Res. J., 2011; 18, 373–380. [18] N.P. Laxmi, M.A. Mutamed, P.S. Nagendra. Int. Food Res. J., 2011; 18, 445–450. [19] C.-F. Hwang, J.-H. Chang, J.-Y. Houng, C.-C. Tasi, C.-K. Lin, H.-Y. Tsen. Biotechnol. Bioprocess Eng., 2012; 17, 827–834. [20] S.K. Akolkar, A.D. Sajgure, S.S. Lele. Indian J. Biotechnol., 2006; 5, 184–188. [21] T. Vasiljevic, P. Jelen. Innov. Food Sci. Emerg, 2001; 2(2), 75–85. [22] R. Rech, M.A.Z. Ayub. Process Biochem., 2007; 42, 873–877. [23] B.L.B. Perini, H.C.M. Souza, M. Kelbert, G.P. Apati, A.P.T. Pezzin, A.L.S. Schneider. Chem. Eng. Trans., 2013; 32, 991–996. [24] H.S. Oberoi, S. Bansal, G.S. Dhillon. Int. J. Food Sci. Tech., 2008; 43, 1499–1504. [25] R. Pinheiro, I. Belo, M. Mota. Lett. Appl. Microbiol., 2003; 37, 438–442. [26] A.M. Gupte, J.S. Nair. J. Sci. Ind. Res. India, 2010; 69, 855–859. [27] A.P. Menera, J.D.C. Ores, V.A. Ribeiro, C.A.V. Burkert, S.J. Kalil. Food Technol. Biotechnol., 2008; 46(1), 66–72. [28] S. Bansal, H.S. Oberoi, G.S. Dhillon, R.T. Patil. Indian J. Microbiol., 2008; 48, 337–341. © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
[29] S. Barberies, R. Segovia. J. Chem. Technol. Biotechnol., 2002; 77, 706–710. [30] R. Rech, M.Z. Ayub. Brazil. J. Chem. Eng., 2006; 23(4), 435–442. [31] P. Guimaraes, J. Klein, L. Domingues, J.A. Teixeira. Braz. J. Food Technol., 2005, 34–39. [32] S.A.D. Bales, F.J. Castillo. Appl. Environ. Microb., 1979; 37(6), 1201–1205. [33] X. Li, J.W. Robbins Jr., K.B. Taylor. J. Ind. Microbiol., 1990; 5, 85–94. [34] A. Jokar, A. Karbassi. J. Agr. Sci. Tech., 2011; 13, 577–584. [35] D.J.M. Kumar, M. Sudha, S. Devika, M.D. Balakumaran, M.R. Kumar, P.T. Kalaichelvan. Ann. Biol. Res., 2012; 3(4), 1712–1718. [36] M. Asif, M.T. Khan, S.A. Rasool. J. Chem. Soc. Pak, 2009; 31(2), 329–334. [37] N. Akcan. Afr. J. Microbiol. Res., 2011; 5(26), 4615–4621. [38] M.V.R. Rao, S.M. Dutta. Appl. Environ. Microb., 1977; 34(2), 185–188. [39] N. Batra, J. Sing, U.C. Banarjee, P.R. Patnaik, R.C. Sobti. Biotechnol. Appl. Biochem., 2002; 36, 1–6. [40] A. Pavani, M.S. Gadge, T. Prabhakar, P. Rupesh. Asian J. Exp. Biol. Sci., 2011; 2(1), 23–27. [41] Z. Nagy, T. Kiss, A. Szentirmai, S. Biro. Protein Expres. Purif., 2011; 21, 24–29. [42] S. Mirdamadi, N. Moazami, M.N. Gorgani. Sci. I. R. Iran., 1997, 23–27. [43] S. Nizamuddin, A. Sridevi, G. Narasimha. Afr. J. Biotechnol., 2008; 7(8), 1096–1100. [44] S.A. Shaikh, J.M. Khire, M.I. Khan. J. Ind. Microbiol. Biot., 1997; 19, 239–245. [45] L. Domingues, N. Lima, J.A. Teixeira. Process Biochem., 2005; 40, 1151–1154. [46] M.E.N.D. Macías, M.C.M.D. Nadra, A.M.S.D. Saad, A.A.P.D.R. Holgado, G. Oliver. Curr. Microbiol., 1983; 9, 99–103. [47] S. Chakraborti, R.K. Sani, U.C. Banerjee, R.C. Sobti. J. Ind. Microbiol. and Biotechnol., 2000; 24(1), 58–63. [48] T.H. Nguyen, B. Splechtna, M. Steinböck, W. Kneifel, H.P. Lettner, K.D. Kulbe, D. Haltrich. J. Agric. Food Chem., 2006; 54(14), 4989–4998. [49] A. Vetere, S. Paoletti. Biochim. Biophys. Acta, 1998; 1380, 223–231. [50] H. Fujimoto, M. Miyasato, Y. Ito, T. Sasaki, K. Ajisaka. Glycoconj. J., 1998; 15, 155–160. [51] G.A. McFeters, W.E. Sandine, P.R. Elliker. J. Bacteriol., 1967; 93(3), 914–919. [52] N.A. Greenberg, R.R. Mahoney. J. Food Sci., 1982; 47, 1824–1835. [53] W. Chen, H. Chen, Y. Xia, J. Zhao, F. Tian, H. Zhang. J. Dairy Sci., 2008; 91, 1751–1758. [54] Y. Li, H. Wang, L. Lu, Z. Li, X. Xu, M. Xiao. Appl. Biochem. Biotechnol., 2009; 158, 192–199. [55] O. Juajun, T.H. Nguyen, T. Maischberger, S. Iqbal, D. Haltrich, M. Yamabhai. Appl. Microbiol. Biotechnol., 2011; 89, 645–654. [56] A.M. Biaikowska, H. Ciemlijski, K.M. Nowakowska, J. Kur, M. Turkiewicz. Arch. Microbiol., 2009; 191, 825–835. [57] C.A. Hsu, R.C. Yu, C.C. Chou. World J. Microbiol. Biotechnol., 2006; 22(4), 355–361. [58] V. Dumortier, C. Brassart, S. Bouquelet. Biotechnol. Appl. Biochem., 1994; 19, 341–354. [59] M.N. Hung, B.H. Lee. Appl. Microbiol. Biotechnol., 2002; 58, 439–445. [60] K. Ohtsuka, A. Tanoh, O. Ozawa, T. Kanematsu, T. Uchida, R. Shinke. J. Ferment. Bioeng., 1990; 70, 301–307. [61] Y. Tanaka, A. Kagamiishi, A. Kiuchi, T. Horiuchi. J. Biochem., 1975; 77, 241–247. [62] D. Todorova-Balvay, I. Stoilova, S. Gargova, M.A. Vijayalakshmi. J. Mol. Recognit., 2006; 19, 299–304. [63] F. Widmer, J.L. Leuba. Eur. J. Biochem., 1979; 100, 559–567. [64] W.H.M. van Casteren, M. Eimermann, L.A.M. van den Broek, J.P. Vincken, H.A. Schols, A.G.J. Voragen. Carbohydr. Res., 2000; 329, 75–85. [65] M. Becerra, E. Cerdán, M.I. González Siso. Biol. Proced. Online, 1998; 1, 48–58. Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
Asia-Pacific Journal of Chemical Engineering [66] M. Becerra, E. Cerdan, M.I. Gonzalez Siso. Biotechnol. Techniques, 1998; 12, 253–256. [67] R.C. Dickson, L.R. Dickson, J.S. Markin. J. Bacteriol., 1979; 137, 51–61. [68] J.A. Goncalves, F.J. Castillo. J. Dairy Sci., 1982; 65, 2088–2094. [69] L. Biermann, M.D. Glantz. Biochim. Biophys. ActaEnzymology, 1968; 167, 373–377. [70] M. Nakao, M. Hatada, Y. Kodama, T. Nakayama, Y. Shibano, T. Amachi. Appl. Microbiol. Biotechnol., 1994; 40, 657–663. [71] N. Onishi, T. Tanaka. Appl. Environ. Microbiol., 1995; 61, 4026–4030. [72] J.L. Berger, B.H. Lee, C. Lacroix. Biotechnol. Appl. Biochem., 1997; 25, 29–41. [73] G.P. Dimri, X. Lee, G. Basile, M. Acosta, G. Scott, C. Roskelley, E.E. Medrano, M. Linskens, I. Rubelj, O.P. Smith. Proc. Natl. Acad. Sci., 1995; 92(20), 9363–9367. [74] Q.Z.K. Zhou, X.D. Chen. Biochem. Eng. J., 2001; 9, 33–40. [75] M.A. Boon, A.E.M. Janssen, K.V. Riet. Enzyme Microb. Technol., 2000; 26, 271–281. [76] M. Harju. Bull. Int. Dairy Fed., 1987; 212, 50–54. [77] E. Jurado, F. Camacho, G. Luzón, J.M. Vicaria. Enzyme Microb. Technol., 2002; 31, 300–309. [78] M. Madani, M. R. Zarif-fard, F. Golchin-far. Arch. Razi. Ins., 1999; 50, 93–98. [79] T. Nakagawa, R. Ikehata, T. Myoda, T. Miyaji, N. Tomizuka. Protein Exp. Purif., 2007; 54, 295–299. [80] S.W.A. Hinz, L.A.M.V.D. Brock, G. Beldman, J.P. Vincken, A.G.J. Voragen. Appl. Microbiol. Biotechnol., 2004; 66, 276–284. [81] L. Lu, M. Xiao, X. Xu, Z. Li, Y. Li. Biochem. Biophys. Res. Commun., 2007; 356, 78–84. [82] T.H. Nguyen, B. Splechtna, S. Krasteva, W. Kneifel, K.D. Kulbe, C. Divne, D. Haltrich. FEMS Microbiol. Lett., 2007; 269, 136–144. [83] J.K. Jeong, O. Kwon, Y.M. Lee, D.B. Oh, J.M. Lee, S. Kim, E.H. Kim, T.N. Le, D.K. Rhee, H.A. Kang. J. Bacteriol., 2009; 191, 3011–3023. [84] B.D. Lauro, A. Strazzulli, G. Perugino, F.L. Cara, E. Bedini, M.M. Corsaro, M. Rossi, M. Moracci. Biochim. Biophys. Acta, 2008; 1784, 292–301. [85] A.R. Park, D.K. Oh. Appl. Microbiol. Biotechnol., 2010; 85(5), 1427–1435. [86] G. Placier, H. Watzlawick, C. Rabiller, R. Mattes. Appl. Environ. Microbiol., 2009; 75(19), 6312–6321. [87] C.S. Kim, E.S. Ji, D.K. Oh. J. Appl. Microbiol., 2004; 97, 1006–1014. [88] M. Ladero, A. Santos, J.L. Garcia, A.V. Carrascosa, B.C.C. Pessela, F. Garcia-Ochoa. Enzyme Microb. Technol., 2002; 30, 392–405. [89] C.S. Kim, E.S. Ji, D.K. Oh. Biotechnol. Lett., 2003; 25, 1769–1774. [90] E. Ishikawa, T. Sakai, H. Ikemura, K. Matsumoto, H. Abe. J. Biosci. Bioeng., 2005; 99, 331–339. [91] Y.J. Cho, H.J. Shin, C. Bucke. Biotechnol. Lett., 2003; 25, 2107–2111. [92] F.M. Pisani, R. Rella, C.A. Raia, C. Rozzo, R. Nucci, A. Gambacorta, M.D. Rosa, M. Rossi. Eur. J. Biochem., 1990; 187, 321–328. [93] A.V. Fowler, I. Zabin. J. Biol. Chem., 1970; 245(19), 5032–5041. [94] R.H. Jacobson, X.J. Zhang, R.F. Dubose, B.W. Matthews. Nature, 1994; 369(6483), 761–766. [95] J.C. Gebler, R. Aebersold, S.G. Withers. J. Biol. Chem., 1992; 267(16), 11126–11130. [96] I.Y. Volkov, N.A. Lunina, O.V. Berezina, G.A. Velikodvorskaya, V.V. Zverlov. Mol. Biol., 2005; 39(6), 799–805. [97] G.K. Shoemaker, D.H. Juers, J.M. Coombs, B.W. Matthews, D.B. Craig. Biochemistry-US, 2003; 42(6), 1707–1710. [98] E.R. Nichols, J.M.A. Gavina, R.G. McLeod, D.B. Craig. Protein J., 2007; 26(2), 95–105. [99] E.R. Nichols, D.B. Craig. Protein J., 2008; 27, 376–383. [100] D.B. Craig, E.R. Nichols. Electrophoresis, 2008; 29, 4298–4303. © 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
β-GALACTOSIDASE: A REVIEW [101] B.C.C. Pessela, R. Fernández-Lafuente, M. Fuentes, A. Vián, J.L. Garcı́a, A.V. Carrascosa, C. Mateo, J.M. Guisán. Enzyme Microb. Tech., 2003; 32(3–4), 369–374. [102] B.C. Pessela, C. Mateo, M. Fuentes, A. Vian, J.L. García, A.V. Carrascosa, J.M. Guisán, R. Fernández-Lafuente. Biotechnol. Prog., 2004; 20(1), 388–392. [103] B.C. Pessela, G. Dellamora-Ortiz, L. Betancor, M. Fuentes, J.M. Guisán, R. Fernandez-Lafuente. Enzyme Microb. Tech, 2007; 40, 310–315. [104] Y. Numanoğlu, S. Sungur. Process Biochem., 2004; 39, 703–709. [105] A. Goel, R.K. Sharma, H.K.L. Tandon. J. Food. Sci. Technol., 2006; 43(5), 526–531. [106] Z. Grosová, M. Rosenberg, M. Gdovin, L. Sláviková, M. Rebroš. Food Chem., 2009; 116, 96–100. [107] R. Torres, C. Mateo, G. Fernández-Lorente. Biotechnol. Prog., 2003; 19(3), 1056–1060. [108] M. Portaccio, S. Stellato, S. Rossi. Enzyme Microb. Tech., 1998; 23(1-2), 101–106. [109] C. Bucke. J. chemical technol. Biotechnol, 1989; 46, pp. 162–163. http://onlinelibrary.wiley.com/doi/10.1002/ jctb.v46:2.n/issuetoc [110] S.T. Yang, M.R. Okos. Biotechnol. Bioeng., 1989; 33(7), 873–885. [111] A.P. Bakken, C.G. Hill, C.H. Amundson. Biotechnol. Bioeng., 1990; 36(3), 293–309. [112] H. Hronska, M. Rosenberg, Z. Grosová. New Biotechnol., 2009; 25, 13–16. [113] M.P. Mariotti, H. Yamanaka, A.R. Araujo, H.C. Trevisan. Braz. Arch. Biol. Techn., 2008; 51(6), 1233–1240. [114] A. Kozhukharova, N. Kirova, K. Batsolva, S. Gargova, A. Dimova. Khranit. Promst., 1991; 40, 43–46. [115] A. Rossi, A. Morana, I. Di Lernia, A. Ottombrino, M. De Rosa. Ital. J. Biochem., 1999; 48(2), 91–97. [116] R. Gaur, H. Pant, R. Jain, S.K. Khare. Food Chem., 2006; 97(3), 426–430. [117] N. Albayrak, S.T. Yang. Enzyme Microb. Tech., 2002; 31(4), 371–383. [118] N. Papayannakos, G. Markas, D. Kekos. The Chem. Eng., 1993; 52(1), B1–B12. [119] D.F.M. Neri, V.M. Balcão, M.G. Carneiro-da-Cunha, L.B. Carvalho, J.A. Teixeira. Catal. Commu., 2008; 9(14), 2334–2339. [120] M. Decleire, W. De Cat, N. Van Huynh, J.C. Motte. Acta Biotechnol., 1987; 7(6), 563–566. [121] I. Roy, M.N. Gupta. Process Biochem., 2003; 39(3), 325–332. [122] C.R. Carrara, A.C. Rubiolo. Chem. Eng. J., 1997; 65(2), 93–98. [123] C.R. Carrara, A.C. Rubiolo. Biotechnol. Prog., 1994; 10(2), 220–224. [124] X. Li, Q.Z.K. Zhou, X.D. Chen. Chem. Eng. Proc: Proc Inten, 2007; 46(5), 497–500. [125] C. Giacomini, A. Villarino, L. Franco-Fraguas, F. BatistaViera. J. Mol. Catal-B Enzym., 1998; 4(5-6), 313–327. [126] B. Champluvier, B. Kamp, P.G. Rouxhet. App. Microbiol. Biot., 1988; 27(5-6), 464–469. [127] M.I. González Siso, S. Suárez Doval. Enzyme Microb. Tech., 1994; 16(4), 303–310. [128] K. Ovsejevi, V. Grazú, F. Batista-Viera. Biotechnol. Tech., 1998; 12(2), 143–148. [129] E. Taqieddin, M. Amiji. Biomaterials, 2004; 25(10), 1937–1945. [130] M. Ladero, A. Santos, F. García-Ochoa. Enzyme Microb. Tech., 2000; 27(8), 583–592. [131] J.L. Berger, B.H. Lee, C. Lacroix. Biotechnol. Tech., 1995; 9(8), 601–606. [132] B.C.C.H. Pessela, C. Mateo, M. Fuentes. Enzyme Microb. Tech., 2003; 33(2-3), 199–205. [133] Z. Li, M. Xiao, L. Lu, Y. Li. Process Biochem., 2008; 43(8), 896–899. [134] M. Banerjee, A. Chakrabarty, S.K. Majumdar. Biotechnol. Bioeng., 1982; 24(8), 1839–1850. [135] A. Dwevedi, A.M. Kayastha. Bioresour. Technol., 2009; 100(10), 2667–2675. [136] H.P.S. Makkar, O.P. Sharma. Biotechnol. Bioeng., 1983; 25, 592–597. Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
347
348
A. NATH ET AL. [137] M. Di Serio, C. Maturo, E. De Alteriis, P. Parascandola, R. Tesser, E. Santacesaria. Catal. Today, 2003; 79-80, 333–339. [138] A.P. Bakken, C.G. Hill, C.H. Amundson. Biotechnol. Bioeng., 1992; 39(4), 408–417. [139] M.J. Hernaiz, D.H.G. Crout. Enzyme Microb. Tech., 2000; 27(1-2), 26–32. [140] S.K. Khare, M.N. Gupta. Biotechnol. Bioeng., 1988; 31(8), 829–833. [141] O. Ariga, M. Kato, T. Sano, Y. Nakazawa, Y. Sano. J. Ferment. Bioeng., 1993; 76(3), 203–206. [142] A. Bódalo, E. Gómez, M.F. Máximo, J.L. Gómez, J. Bastida. Biotechnol. Tech., 1991; 5(5), 393–394. [143] V. Kéry, Š. Kučár, M. Matulová, J. Haplová. Carbohyd. Res., 1991; 209(C), 83–87. [144] S.F. D’Souza, R. Kaul, G.B. Nadkarni. Biotechnol. Lett., 1985; 7(8), 589–592. [145] S. Sungur, U. Akbulut. J. Chem. Technol. Biot., 1994; 59(3), 303–306. [146] M. Marrakchi, S.V. Dzyadevych, F. Lagarde, C. Martelet, N. Jaffrezic-Renault. Mater. Sci. Eng. C, 2008; 28, 872–875. [147] S. Campuzano, B. Serra, D. Llull, J.L. Garcia, P. Garcia. Appl. Environ. Microb., 2009; 75(18), 5972–5980. [148] R. Panesar, P.S. Panesar, R.S. Singh, J.F. Kennedy, M.B. Bera. Food Chem., 2007; 101, 786–790. [149] Z. Cui. China Particuology, 2005; 3(6), 343–348. [150] S. Nakao, H. Osada, H. Kurata, T. Tsuru, S. Kimura. Desalination, 1988; 70(1–3), 191–205. [151] S. Bhattacharjee, S. Ghosh, S. Datta, C. Bhattacharjee. Desalination, 2006; 195(1-3), 95–108. [152] P. Sarkar, S. Ghosh, S. Dutta, D. Sen, C. Bhattacharjee. Desalination, 2009; 249, 5–11. [153] S. Bhattacharjee, C. Bhattacharjee, S. Datta. J. Membrane Sci., 2006a; 275(1-2), 141–150. [154] Z. Mlichová, M. Rosenberg. J. Food Nutri. Res., 2006; 45(2), 47–54. [155] G.D. Hatzinikolaoua, E. Katsifas, D. Mamma, A.D. Karagouni, P. Christakopoulosb, D. Kekosb. Biochem. Eng. J., 2005; 24, 161–172. [156] P. Guimaraes, K.J. Domingues, J.A. Teixeira. Braz. J. Food Technol., 2005, 34–39. [157] W.B. Silveira, F.J.V. Passos, H.C. Mantovani, F.M.L. Passos. Enzyme Microb. Tech., 2005; 36(7), 930–936. [158] S. Ozmihci, F. Kargi. Bioresour. Technol., 2007; 98(16), 2978–2984. [159] S. Ozmihci, F. Kargi. Bioprocess Biosyst. Eng., 2007; 30(2), 79–86. [160] M. Rosenberg, M. Tomáška, J. Kaňuch, E. Šurdík. Acta Biotechnol., 1995; 15(4), 387–390. [161] A.C. Silva, P.M.R. Guimares, J.A. Teixeir, L. Domingues. J. Ind. Microbiol. Biot., 2010; 37(9), 973–982. [162] M.K. Tahoun, T.M. El-Nemr, O.H. Shata. Eur. Food Res. Technol., 1999; 208(1), 60–64. [163] S. Ozmihci, F. Kargi. Biochem. Eng. J., 2008; 42, 180–185. [164] M.A. Mehaia, M. Cheryan. Bioprocess Eng., 1990; 5(2), 57–61. [165] M. Staniszewski, W. Kujawski, M. Lewandowska. J. Food Eng., 2009; 91, 240–249. [166] A. Gosling, G.W. Stevens, A.R. Barber, S.E. Kentish, S.L. Gras. Food Chem., 2010; 121(2), 307–318. [167] M.A. Boon, A.E.M. Janssen, A.V.D. Padt. Biotechnol. Bioeng., 1999; 64(5), 558–567. [168] D.F.M. Neri, V.M. Balcão, R.S. Costa, I.C.A.P. Rocha, E. M.F.C. Ferreira, D.P.M. Torres, L.R.M. Rodrigues, J.L.B. Carvalho, J.A. Teixeira. Food Chem., 2009; 115(1), 92–99.
© 2014 Curtin University of Technology and John Wiley & Sons, Ltd.
Asia-Pacific Journal of Chemical Engineering [169] T. Palai, S. Mitra, P.K. Bhattacharya. J. Bios. Bioeng., 2012; 114(4), 418–423. [170] S.J. Jung, R. Houde, B. Baurhoo, X. Zhao, B.H. Lee. Poult. Sci., 2008; 87(9), 1694–1699. [171] R.C. Martinez, H.R. Cardarelli, W. Borst, S. Albrecht, H. Schols, O.P. Gutiérrez, A.J. Maathuis, B.D.D.M. Franco, E.C.D. Martinis, E.G. Zoetendal, K. Venema, S.M. Saad, H. Smidt. FEMS Microbiol. Ecol., 2013; 84(1), 110–123. [172] K. Iwasaki, M. Nakajima, S. Nakao. Process Biochem., 1996; 31, 69–76. [173] Z. Mozaffar, K. Nakanishi, R. Matsuno. J. Food Sci., 1985; 50, 1602–1606. [174] C.A. Hsu, S.L. Lee, C.C. Chou. J. Agric. Food Chem., 2007; 55, 2225–2230. [175] B. Splechtna, T.H. Nguyen, M. Steinbock, K.D. Kulbe, W. Lorenz, D. Haltrich. J. Agric. Food Chem., 2006; 54, 4999–5006. [176] P. Nakkharat, K.D. Kulbe, M. Yamabhai, D. Haltrich. Biotechnol. J., 2006, 1,633–1,638. [177] R. Cruz, V.D.A. Cruz, J.G.B.D. Oliveira, M. Khenayfes, C. Dorta, L.H.D.S. Oliveira, E. Ardiles, A. Galli. Bioresour. Technol., 1999; 70, 165–171. [178] H.Y. Park, H.J. Kim, J.K. Lee, D. Kim, D.K. Oh. World J. Microbiol. Biotechnol., 2008; 24, 1553–1558. [179] E.S. Ji, N.H. Park, D.K. Oh. World J. Microbiol. Biotechnol., 2005; 21, 759–764. [180] G. Tzortzis, A.K. Goulas, G.R. Gibson. Appl. Microbiol. Biotechnol., 2005; 68, 412–416. [181] N. Onishi, K. Yokozeki. J. Ferment., Bioeng., 1996; 82, 124–127. [182] P. Zheng, H. Yu, Z. Sun, Y. Ni, W. Zhang, Y. Fan, Y. Xu. Biotechnol. J., 2006, 1,1464–1,1470. [183] Z. Mozaffar, K. Nakanishi, R. Matsuno. Appl. Microbiol. Biotechnol., 1986; 25, 229–231. [184] H.J. Shin, J.W. Yang. J. Microbiol. Biotechnol., 1998, 8,484–8,489. [185] S. Chockchaisawasdee, V.I. Athanasopoulos, K. Niranjan, R.A. Rastall. Biotechnol. Bioeng., 2004; 89, 434–443. [186] P. Nakkharat, K.D. Kulbe, M. Yamabhai, D. Haltrich. Biotechnol. J., 2006; 1, 633–638. [187] O. Ozawa, K. Ohtsuka, T. Uchida, N.S.K. Gakkaishi. Nippon Shokuhin Kogyou Gakkaish s, 1989; 36, 898–902. [188] M.J. In, H.J. Chae. Korean J. Appl. Microbiol. Biothechnol., 1998; 26, 40–44. [189] M.E. Bruins, M. Strubel, J.F.T.V. Lieshout, A.E.M. Janssen, R.M. Boom. Enzyme Microb. Technol., 2003; 33, 3–11. [190] K. Akiyama, M. Takase, K. Horikoshi, S. Okonogi. Biosci. Biotechnol. Biochem., 2001; 65, 438–441. [191] N. Onishi, I. Kira, K. Yokozeki. Lett. Appl. Microbiol., 1996; 23, 253–256. [192] T. Sakai, H. Tsuji, S. Shibata, K. Hayakawa, K. Matsumoto. J. Gen. Appl. Microbiol., 2008; 54, 285–293. [193] K.M. Anderson, H. Ashida, K. Maskos, A. Dell, S.C. Li, Y. T. Li. J. Biolog. Chem., 2005; 280(9), 7720–7728. [194] T. Kobayashi, D. Liu, H. Ogawa, Y. Miwa, T. Nagasaka, S. Maruyama, Y. Li, A. Onishi, M. Iwamoto, T. Kuzuya, K. Kadomatsu, K. Uchida, A. Nakao. Transpl. Immunol., 2009; 20, 132–138. [195] A.B. Liu, S.R. Roffler. Tzu. Chi. Med. J., 2006; 18(4), 259–265. [196] H. Wang, H. Luo, Y. Bai, Y. Wang, P. Yang, P. Shi, W. Zhang, Y. Fan, B. Yao. J. Agric. Food Chem., 2009; 57, 5535–5541. [197] M.T. Sanchez-Aparicio, M.F. Rosas, R.M. Ferraz, L. Delgui, J.J. Veloso, E. Blanco, A. Villaverde, F. Sobrino. Clin. Vaccine Immunol., 2009; 16(8), 1228–1235.
Asia-Pac. J. Chem. Eng. 2014; 9: 330–348 DOI: 10.1002/apj
