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Enzymatic production of lactulose and 1-lactulose: current state and perspectives

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Abstract

Lactulose, a synthetic ketose disaccharide, has been widely used in food and pharmaceutical industries as prebiotic food additives and drugs against constipation and hepatic encephalopathy. Lactulose has, so far, been produced chemically using lactose on a commercial scale. The key problems associated with such chemical process are the cost of removal of the catalyst and colored by-products and the product safety. Enzymatic production of lactulose is safe, environment-friendly, and simpler in comparison to the chemical method. As a promising alternative to the chemical method, enzymatic conversion of lactose into lactulose by β-galactosidase or cellobiose 2-epimerase has recently gained a great deal of attention. This could be considered as a possible route for whey surplus because lactose is the major component of cheese whey. Herein, we summarize recent advances on the enzymatic synthesis of lactulose with emphasis on the selectivity of biocatalysts and their catalytic efficiency in free and immobilized forms. The production of 1-lactulose by enzymatic bioconversion of lactose has also been discussed. Furthermore, future research needs of β-galactosidase and cellobiose 2-epimerase for the enzymatic synthesis of lactulose and 1-lactulose are reviewed.

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References

  • Adachi S, Patton S (1961) Presence and significance of lactulose in milk products: a review. J Dairy Sci 44:1375–1393

    Article  CAS  Google Scholar 

  • Adamczak M, Charubin D, Bednarski W (2009) Influence of reaction medium composition on enzymatic synthesis of galactooligosaccharides and lactulose from lactose concentrates prepared from whey permeate. Chem Pap 63:111–116

    Article  CAS  Google Scholar 

  • Agustín O, Nieves C (2009) Lactulose as a food ingredient. J Sci Food Agric 89:1987–1990

    Article  Google Scholar 

  • Aider M, Halleux D (2007) Isomerization of lactose and lactulose production: review. Trends Food Sci Technol 18:356–364

    Article  CAS  Google Scholar 

  • Ajisaka K, Fujimoto H, Nishida H (1988) Enzymic synthesis of disaccharides by use of the reversed hydrolysis activity of β-d-galactosidases. Carbohydr Res 180:35–42

    Article  CAS  Google Scholar 

  • Ajisaka K, Nishida H, Fujimoto H (1987) Use of an activated carbon column for the synthesis of disaccharides by use of a reversed hydrolysis activity of β-galactosidase. Biotechnol Lett 9:387–392

    Article  CAS  Google Scholar 

  • Anderson NG (2012) Using continuous processes to increase production. Org Process Res Dev 16:852–869

    Article  CAS  Google Scholar 

  • Athanasiadis I, Boskou D, Kanellaki M, Kiosseoglou V, Koutinas AA (2002) Whey liquid waste of the dairy industry as raw material for potable alcohol production by kefir granules. J Agric Food Chem 50:7231–7234

    Article  CAS  Google Scholar 

  • Bednarski W, Kulikowska A (2007) Influence of two-phase system composition on biocatalytic properties of β-galactosidase preparations. Chem Pap 61:364–369

    Article  CAS  Google Scholar 

  • Bhosale SH, Rao MB, Deshpande VV (1996) Molecular and industrial aspects of glucose isomerase. Microbiol Rev 60:280–300

    CAS  Google Scholar 

  • Boller T, Meier C, Menzler S (2002) Eupergit oxirane acrylic beads: how to make enzymes fit for biocatalysts. Org Process Res Dev 6:509–519

    Article  CAS  Google Scholar 

  • Bolto B, Dixon D, Eldridge R, King S, Linge K (2002) Removal of natural organic matter by ion exchange. Water Res 36:5057–5065

    Article  CAS  Google Scholar 

  • Bornscheuer UT, Huisman GW, Kazlauskas RJ, Lutz S, Moore JC, Robins K (2012) Engineering the third wave of biocatalysis. Nature 485:185–194

    Article  CAS  Google Scholar 

  • Bourne Y, Henrissat B (2001) Glycoside hydrolases and glycosyltransferases: families and functional modules. Curr Opin Struct Biol 11:593–600

    Article  CAS  Google Scholar 

  • Chen X, Zhai X, Kang Z, Sun X (2012) Lactulose: an effective preventive and therapeutic option for ischemic stroke by production of hydrogen. Med Gas Res 2:3

    Article  CAS  Google Scholar 

  • Chen X, Zuo Q, Hai Y, Sun X (2011) Lactulose: an indirect antioxidant ameliorating inflammatory bowel disease by increasing hydrogen production. Med Hypotheses 76:325–327

    Article  CAS  Google Scholar 

  • Dagbagli S (2008) Optimization of β-galactosidase production using Kluyveromyces lactis NRRL Y-8279 by response surface methodology. Electron J Biotechnol 11:1–12

    Google Scholar 

  • Davies G, Henrissat B (1995) Structures and mechanisms of glycosyl hydrolases. Structure 3:853–859

    Article  CAS  Google Scholar 

  • De Muynck C, Beauprez J, Soetaert W, Vandamme EJ (2006) Boric acid as a mobile phase additive for high performance liquid chromatography separation of ribose, arabinose and ribulose. J Chromatogr A 1101:115–121

    Article  Google Scholar 

  • Deschavanne PJ, Viratelle OM, Yon JM (1978) Conformational adaptability of the active site of β-galactosidase. J Biol Chem 253:833–837

    CAS  Google Scholar 

  • De Souza Oliveira RP, Rodrigues Florence AC, Perego P, De Oliveira MN, Converti A (2011) Use of lactulose as prebiotic and its influence on the growth, acidification profile and viable counts of different probiotics in fermented skim milk. Int J Food Microbiol 145:22–27

    Article  Google Scholar 

  • European Commission (2011) Commission regulation (EC) No. 575/2011 of 16 June 2011 on the catalogue of feed materials. Off J Europ Union L 159:25–65

    Google Scholar 

  • Fischer L, Bromann R, Kengen SWM, de Vos WM, Wagner F (1996) Catalytical potency of β-glucosidase from the extremophile Pyrococcus furiosus in glucoconjugate synthesis. Biotechnology 14:88–91

    Article  CAS  Google Scholar 

  • Förster-Fromme K, Schuster-Wolff-Bühring R, Hartwig A, Holder A, Schwiertz A, Bischoff CS, Hinrichis J (2011) A new enzymatically produced 1-lactulose: a pilot study to test the bifidogenic effects. Int Dairy J 21:940–948

    Article  Google Scholar 

  • Foster KJ, Lin S, Turck CJ (2010) Current and emerging strategies for treating hepatic encephalopathy. Crit Care Nurs Clin North Am 22:341–350

    Article  Google Scholar 

  • Fujiwara T, Saburi W, Inoue S, Mori H, Matsui H, Tanaka I (2013) Crystal structure of Ruminococcus albus cellobiose 2-epimerase: structural insights into epimerization of unmodified sugar. FEBS Lett 587:840–846

    Article  CAS  Google Scholar 

  • Gänzle MG, Haase G, Jelen P (2008) Lactose: crystallization, hydrolysis and value-added derivatives. Int Dairy J 18:685–694

    Article  Google Scholar 

  • Guerrero C, Vera C, Plou F, Illanes A (2011) Influence of reaction conditions on the selectivity of the synthesis of lactulose with microbial β-galactosidases. J Mol Catal B: Enzym 72:206–212

    Article  CAS  Google Scholar 

  • Guimarães PM, Teixeira JA, Domingues L (2010) Fermentation of lactose to bio-ethanol by yeasts as part of integrated solutions for the valorisation of cheese whey. Biotechnol Adv 28:375–384

    Article  Google Scholar 

  • Hanssen T, Andersson M, Wehtje E, Adlercreutz P (2001) Influence of water activity on the competition between β-galactosidase-catalysed transglycosylation and hydrolysis. Enzyme Microb Technol 29:527–534

    Article  Google Scholar 

  • He X, Zhang S, Yang S (2001) Cloning and expression of thermostable β-glycosidase gene from Thermus nonproteolyticus HG102 and characterization of recombinant enzyme. Appl Biochem Biotechnol 94:243–255

    Article  CAS  Google Scholar 

  • Henrissat BA (1991) A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 280:309–316

    CAS  Google Scholar 

  • Hicks KB, Parrish FW (1980) A new method for the preparation of lactulose from lactose. Carbohydr Res 82:393–397

    Article  CAS  Google Scholar 

  • Hua X, Yang R, Zhang W, Fei Y, Jin Z, Jiang B (2010) Dual-enzymatic synthesis of lactulose in organic-aqueous two-phase media. Food Res Int 43:716–722

    Article  CAS  Google Scholar 

  • Hua X, Yang R, Shen Q, Ye F, Zhang W, Zhao W (2013) Production of 1-lactulose and lactulose using commercial β-galactosidase from Kluyveromyces lactis in the presence of fructose. Food Chem 137:1–7

    Article  CAS  Google Scholar 

  • Ito S (2009) Features and applications of microbial sugar epimerases. Appl Microbiol Biotechnol 84:1053–1060

    Article  CAS  Google Scholar 

  • Ito S, Taguchi H, Hamada S, Kawauchi S, Ito H, Senoura T, Watanabe J, Nishimukai M, Ito S, Matsui H (2008) Enzymatic properties of cellobiose 2-epimerase from Ruminococcus albus and the synthesis of rare oligosaccharides by the enzyme. Appl Microbiol Biotechnol 79:433–441

    Article  CAS  Google Scholar 

  • Juajun O, Nguyen TH, Maischberger T, Iqbal S, Haltrich D, Yamabhai M (2010) Cloning, purification, and characterization of β-galactosidase from Bacillus licheniformis DSM 13. Appl Microbiol Biotechnol 89:645–654

    Article  Google Scholar 

  • Katchalski-Katzir E, Kraemer DM (2000) Eupergit® C, a carrier for immobilization of enzymes of industrial potential. J Mol Catal B: Enzym 10:157–176

    Article  CAS  Google Scholar 

  • Kim NH, Kim HJ, Kang DI, Jeong KW, Lee JK, Kim Y, Oh DK (2008) Conversion shift of d-fructose to d-psicose for enzyme-catalyzed epimerization by addition of borate. Appl Environ Microbiol 74:3008–3013

    Article  CAS  Google Scholar 

  • Kim YS, Kim JE, Oh DK (2013) Borate enhances the production of lactulose from lactose by cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus. Bioresour Technol 128:809–812

    Article  CAS  Google Scholar 

  • Kim YS, Oh DK (2012) Lactulose production from lactose as a single substrate by a thermostable cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus. Bioresour Technol 104:668–672

    Article  CAS  Google Scholar 

  • Kim YS, Park CS, Oh DK (2006) Lactulose production from lactose and fructose by a thermostable β-galactosidase from Sulfolobus solfataricus. Enzyme Microb Technol 39:903–908

    Article  CAS  Google Scholar 

  • Kumari S, Panesar PS, Panesar R (2011a) Production of β-galactosidase using novel yeast isolate from whey. Int J Dairy Sci 6:150–157

    Article  CAS  Google Scholar 

  • Kumari S, Panesar PS, Bera MB, Singh B (2011b) Permeabilization of yeast cells for β-galactosidase activity using mixture of organic solvents: a response surface methodology approach. Asian J Biotechnol 3:406–411

    Article  Google Scholar 

  • Lee YJ, Kim CS, Oh DK (2004) Lactulose production by β-galactosidase in permeabilized cells of Kluyveromyces lactis. Appl Microbiol Biotechnol 64:787–793

    Article  CAS  Google Scholar 

  • Lilly M, Dunnill P (1976) Immobilized-enzymes reactors. In: Mosbach K (ed) Methods in enzymology, vol. 44. Academic, New York, pp 717–738

    Google Scholar 

  • Lindmark-Mansson L, Fonden R, Pettersson HE (2003) Composition of Swedish dairy milk. Int Dairy J 13:409–425

    Article  CAS  Google Scholar 

  • Lorenzen PC, Breiter J, Clawin-Radecker I, Dau A (2013) A novel bi-enzymatic system for lactose conversion. Int J Food Sci Technol. doi:10.1111/ijfs.12101

    Google Scholar 

  • MacGillivray PC, Finlay HVL, Binns TB (1959) Use of lactulose to create a preponderance of Lactobacilli in the intestine of bottle-fed infants. Scott Med J 4:182–189

    CAS  Google Scholar 

  • Mayer J, Conrad J, Klaiber I, Lutz-Wahl S, Beifuss U, Fischer L (2004) Enzymatic production and complete nuclear magnetic resonance assignment of the sugar lactulose. J Agric Food Chem 52:6983–6990

    Article  CAS  Google Scholar 

  • Mayer J, Kranz B, Fischer L (2010) Continuous production of lactulose by immobilized thermostable β-glycosidase from Pyrococcus furiosus. J Biotechnol 145:387–393

    Article  CAS  Google Scholar 

  • Miletić N, Nastasović A, Loos K (2012) Immobilization of biocatalysts for enzymatic polymerizations: possibilities, advantages, applications. Bioresour Technol 115:126–135

    Article  Google Scholar 

  • Montgomery EM, Hudson CS (1930) Relations between rotatory power and structure in the sugar group. XXVII. Synthesis of a new disaccharide ketose (lactulose) from lactose. J Am Chem Soc 52:2101–2106

    Article  CAS  Google Scholar 

  • Naber JR, Buchwald SL (2010) Packed-bed reactors for continuous-flow C=N cross-coupling. Angew Chem Int Edit 122:9659–9664

    Article  Google Scholar 

  • Olano A, Corzo N (2009) Lactulose as a food ingredient. J Sci Food Agric 89:1987–1990

    Article  CAS  Google Scholar 

  • Panesar PS, Kumari S (2011) Lactulose: production, purification and potential applications. Biotechnol Adv 29:940–948

    Article  CAS  Google Scholar 

  • Park AR, Oh DK (2010) Galacto-oligosaccharide production using microbial β-galactosidase: current state and perspectives. Appl Microbiol Biotechnol 85:1279–1286

    Article  CAS  Google Scholar 

  • Park CS, Kim JE, Choi JG, Oh DK (2011) Characterization of a recombinant cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus and its application in the production of mannose from glucose. Appl Microbiol Biotechnol 92:1187–1196

    Article  CAS  Google Scholar 

  • Pham TT, Shah NP (2008) Effect of lactulose on biotransformation of isoflavone glycosides to aglycones in soymilk by Lactobacilli. J Food Sci 73:158–165

    Article  Google Scholar 

  • Pinelo M, Jonsson G, Meyer AS (2009) Membrane technology for purification of enzymatically produced oligosaccharides: molecular and operational features affecting performance. Sep Purif Technol 70:1–11

    Article  CAS  Google Scholar 

  • Porkka L, Salaminen E, Salaminen S (1988) The effects of lactulose-sweetened yoghurt on the rate of gastric emptying and intestinal transit in healthy human volunteers. Z Ernahrungswiss 27:150–154

    Article  CAS  Google Scholar 

  • Salonen N, Salonen K, Leisola M, Nyysölä A (2013) d-Tagatose production in the presence of borate by resting Lactococcus lactis cells harboring Bifidobacterium longum l-arabinose isomerase. Bioprocess Biosyst Eng 36:489–497

    Article  CAS  Google Scholar 

  • Sanz-Aparicio J, Hermoso JA, Martínez-Ripoll M, Lequerica JL, Polaina J (1998) Crystal structure of β-glucosidase A from Bacillus polymyxa: insights into the catalytic activity in family 1 glycosyl hydrolases. J Mol Biol 275:491–502

    Article  CAS  Google Scholar 

  • Schumann C (2002) Medical, nutritional and technological properties of lactulose. An update. Eur J Nutr 41:I17–I25

    Article  Google Scholar 

  • Seki N, Saito H (2012) Lactose as a source for lactulose and other functional lactose derivatives. Int Dairy J 22:110–115

    Article  CAS  Google Scholar 

  • Sheldon RA (2007) Enzyme immobilization: the quest for optimum performance. Adv Synth Catal 349:1289–1307

    Article  CAS  Google Scholar 

  • Shen Q, Yang R, Hua X, Ye F, Wang H, Zhao W, Wang K (2012) Enzymatic synthesis and identification of oligosaccharides obtained by transgalactosylation of lactose in the presence of fructose using β-galactosidase from Kluyveromyces lactis. Food Chem 135:1547–1554

    Article  CAS  Google Scholar 

  • Siso MI (1996) The biotechnological utilization of cheese whey: a review. Bioresour Technol 57:1–11

    Article  Google Scholar 

  • Song YS, Lee JH, Kang SW, Kim SW (2010) Performance of β-galactosidase pretreated with lactose to prevent activity loss during the enzyme immobilisation process. Food Chem 123:1–5

    Google Scholar 

  • Song YS, Lee HU, Park C, Kim SW (2012a) Batch and continuous synthesis of lactulose from whey lactose by immobilized β-galactosidase. Food Chem 136:689–694

    Article  Google Scholar 

  • Song YS, Shin HY, Lee JY, Park C, Kim SW (2012b) β-galactosidase-immobilised microreactor fabricated using a novel technique for enzyme immobilisation and its application for continuous synthesis of lactulose. Food Chem 133:611–617

    Article  CAS  Google Scholar 

  • Song YS, Lee HU, Park C, Kim SW (2013a) Optimization of lactulose synthesis from whey lactose by immobilized β-galactosidase and glucose isomerase. Carbohydr Res 369:1–5

    Article  CAS  Google Scholar 

  • Song YS, Suh YJ, Park C, Kim SW (2013b) Improvement of lactulose synthesis through optimization of reaction conditions with immobilized β-galactosidase. Korean J Chem Eng 30:160–165

    Article  CAS  Google Scholar 

  • Tang L, Li Z, Dong X, Yang R, Zhang J, Mao Z (2010) Lactulose biosynthesis by β-galactosidase from a newly isolated Arthrobacter sp. J Ind Microbiol Biotechnol 38:471–476

    Article  Google Scholar 

  • Tyler TR, Leatherwood JM (1967) Epimerization of disaccharides by enzyme preparations from Ruminococcus albus. Arch Biochem Biophys 119:363–367

    Article  CAS  Google Scholar 

  • Vaheri M, Kaupinnen V (1978) The formation of lactulose (4-O-β-galactopyranosylfructose) by β-galactosidase. Acta Pharm Fenn 87:75–83

    CAS  Google Scholar 

  • van den Berg R, Peters JA, van Bekkum H (1994) The structure and (local) stability constants of borate esters of mono- and di-saccharides as studied by 11B and 13C NMR spectroscopy. Carbohydr Res 253:1–12

    Article  Google Scholar 

  • Wang K, Lu Y, Liang W, Wang S, Jiang Y, Huang R, Liu Y (2012) Enzymatic synthesis of galacto-oligosaccharides in an organic-aqueous biphasic system by a novel β-galactosidase from a metagenomic library. J Agric Food Chem 60:3940–3946

    Article  CAS  Google Scholar 

  • Wang X, Liu X, Zhao C, Ding Y, Xu P (2011) Biodiesel production in packed-bed reactors using lipase-nanoparticle biocomposite. Bioresour Technol 102:6352–6355

    Article  CAS  Google Scholar 

  • Watanabe H, Yagi M, Nishimoto T, Chaen H, Fukuda S (2010) Cellobiose 2-epimerase, process for producing same, and use of same. Patent WO 2010090095

  • Wollowski I, Rechkammer G, Pool-Zobel BL (2001) Protective role of probiotics and prebiotics in colon cancer. Am J Clinical Nutr 73:451–455

    Google Scholar 

  • Zhang YW, Jeya M, Lee JK (2010) l-Ribulose production by an Escherichia coli harboring l-arabinose isomerase from Bacillus licheniformis. Appl Microbiol Biotechnol 87:1993–1999

    Article  CAS  Google Scholar 

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Acknowledgments

This study was financially supported by the Key Project of National Natural Science Fund (31230057), the National Key Technology R&D Program in the 12th Five Year Plan of China (grant no. 2011BAD23B03), the Natural Science Fund of Jiangsu Province (BK2011149), and Ph.D. Programs Foundation of Jiangnan University (JUDCF09025).

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  1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, Jiangsu, People’s Republic of China

    He Wang

  2. School of Food Science and Technology, Jiangnan University, Wuxi, 214122, Jiangsu, People’s Republic of China

    He Wang, Ruijin Yang, Xiao Hua, Wei Zhao & Wenbin Zhang

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  1. He Wang
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  2. Ruijin Yang
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  3. Xiao Hua
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  4. Wei Zhao
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  5. Wenbin Zhang
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Correspondence to Ruijin Yang.

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Wang, H., Yang, R., Hua, X. et al. Enzymatic production of lactulose and 1-lactulose: current state and perspectives. Appl Microbiol Biotechnol 97, 6167–6180 (2013). https://doi.org/10.1007/s00253-013-4998-3

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