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Lactulose production by β-galactosidase in permeabilized cells of Kluyveromyces lactis

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Abstract

Lactulose production from lactose and fructose was investigated with several commercial β-galactosidases. The enzyme from Kluyveromyces lactis exhibited the highest lactulose productivity among the β-galactosidases tested. The reaction conditions for lactulose production were optimized using cells that had been permeabilized by treatment with 50% (v/v) ethanol: cell concentration, 10.4 g l−1; concentration of substrates, 40% (w/v) lactose and 20% (w/v) fructose; temperature, 60°C; pH 7.0. Under these conditions, the permeabilized cells produced approximately 20 g l−1 lactulose in 3 h with a lactulose productivity of 6.8 g l−1 h−1. These results represent 1.3- and 2.1-fold increases in lactulose concentration and productivity compared with untreated washed cells. This is the first reported trial of enzymatic synthesis of lactulose using permeabilized yeast cells.

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References

  • Burvall A, Asp NG, Dahlqvist A (1979) Oligosaccharide formation during hydrolysis of lactose with Saccharomyces lactis lactase (MaxilactR): part 1-quantitative aspects. Food Chem 4:243–250

    CAS  Google Scholar 

  • Carobbi R, Innocenti F (1990) Process for preparing lactulose from lactose by epimerization with sodium aluminate. European patent 0320670

  • Carobbi R, Miletti S, Franci V (1985) Process for preparing lactulose from lactose, in the form of a syrup or a crystalline product. US Patent 4536221

  • Conn HO, Floch MH (1970) Effect of lactulose and Lactobacillus on the fecal flora. Am J Clin Nutr 23:1588–1615

    PubMed  Google Scholar 

  • Cruz R, Cruz VD, Bolete JG, Khenafes MO, Dorta C, Oliveira LHS, Ardiles E, Galli A (1999) Production of transgalactosylated oligosaccharides (TOS) by galactosyltransferase activity from Penicillium simplicissimum. Bioresour Technol 70:165–171

    Article  Google Scholar 

  • D’Auria S, Nucci R, Rossi M, Bertoli E, Fabio T, Gryczynski I, Malak H, Lakowicz JR (1999) β-Galactosidase from the hyperthermophilic archaeon Sulfolobus solfataricus: structure and activity in the presence of alcohols. J Biochem 126:545–552

    PubMed  Google Scholar 

  • De Harr WT, Pluim H (1991) Method of preparing lactulose. European patent 0339749

  • Deya E, Takahashi K (1991) Production process of high purity lactulose syrup. US Patent 5034064

  • Fontes EAF, Passos FML, Passos FJV (2001) A mechanistical mathematical model to predict lactose hydrolysis by β-galactosidase in a permeabilized cell mass of Kluyveromyces lactis: validity and sensitivity analysis. Process Biochem 37:267–274

    Article  Google Scholar 

  • Gekas V, Lopez-Leiva M (1985) Hydrolysis of lactose: a literature review. Process Biochem 20:2–12

    CAS  Google Scholar 

  • Giacomini C, Irazoqui G, Gonzalez P, Batista-Viera F, Brena BN (2002) Enzymatic synthesis of galactosyl-xylose by Aspergillus oryzae β-galactosidase. J Mol Catal B 19–20:159–165

    Google Scholar 

  • Gowda LR, Bachhwat N, Bhat SG (1991) Permeabilization of baker’s yeast cells by cetytrimethylammonium bromide for intracellular enzyme catalysis. Enzyme Microb Technol 13:154–157

    CAS  Google Scholar 

  • Greenberg NA, Mahoney RR (1983) Formation of oligosaccharides by β-galactosidase from Streptococcus thermophilus. Food Chem 10:195–204

    CAS  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 

  • Hung MN, Lee BH (2002) Purification and characterization of a recombinant β-galactosidase with transgalactosylation activity from Bifidobacterium infantis HL96. Appl Microbiol Biotechnol 58:439–445

    Article  PubMed  Google Scholar 

  • Hung MN, Xia Z, Hu NT, Lee BH (2001) Molecular and biochemical analysis of two β-galactosidases from Bifidobacterium infantis HL96. Appl Environ Microbiol 67:4256–4263

    CAS  PubMed  Google Scholar 

  • Joshi MS, Gowda LR, Kawai LC, Bhat SG (1989) Permeabilization of yeast cells (Kluyeromyces fragilis) to lactose by digitonin. Enzyme Microb Technol 11:439–444

    Article  Google Scholar 

  • Kondo A, Liu Y, Furuta M, Fujita Y, Matsumoto T, Fukuda H (2000) Preparation of high activity whole cell biocatalyst by permeabilization of recombinant flocculent yeast with alcohol. Enzyme Microb Technol 27:806–811

    Article  CAS  PubMed  Google Scholar 

  • Krumbbloz RE, Dorscheid MG (1991) Method of manufacturing lactulose. European patent 0375040

  • Loguercio C, Abbiati R, Rinaldi M, Romano A, Blanco CDV, Coltorti M (1995) Long-term effects of Enterococcus faecium SF68 versus lactulose in the treatment of patients with cirrhosis and grade 1–2 hepatic encephalopathy. J Hepatol 23:39–46

    Article  PubMed  Google Scholar 

  • Méndez A, Olano A (1979) Lactulose: a review on some chemical properties and applications in infant nutrition and medicine. Dairy Sci Abstr 41:531–535

    Google Scholar 

  • Onishi N, Tanaka T (1995) Purification and properties of a novel thermostable galactooligosaccharide from lactose by Sterigmatomyces elviae CBS6803. Appl Environ Microbiol 61:4026–4030

    CAS  PubMed  Google Scholar 

  • Onishi N, Yamashiro A, Yokozeki K (1995) Production of galactooligosaccharide from lactose by Sterigmatomyces elviae CBS6803. Appl Environ Microbiol 61:4022–4025

    CAS  PubMed  Google Scholar 

  • Petzelbauer I, Zeleny R, Reiter A, Kulbe KD, Nidetzky B (2000) Development of an ultra-high-temperature process for the enzymatic hydrolysis of lactose: II. Oligosaccharide formation by two thermostable beta-glycosidases. Biotechnol Bioeng 69:140–149

    Article  PubMed  Google Scholar 

  • Reuter S, Nygaard AR, Zimmermann W (1999) β-Galactooligosaccharide synthesis with β-galactosidases from Sulfolobus solfataricus, Aspergillus oryzae, and Escherichia coli. Enzyme Microb Technol 25:509–516

    Article  Google Scholar 

  • Siso MIG, Cerdán E, Picos MAF (1992) Permeabilization of Kluyveromyces lactis cells for milk whey saccharification: a comparison of different treatments. Biotechnol Tech 6:289–292

    Google Scholar 

  • Voragen AGJ (1998) Technological aspects of functional food-related carbohydrates. Trends Food Sci Technol 9:328–335

    CAS  Google Scholar 

  • Zarate S, Lopez-Leiva MH (1990) Oligosaccharides formation during enzymatic hydrolysis: a review of literature. J Food Prot 53:262–268

    Google Scholar 

  • Zhou QZK, Chen XD (2001) Effects of temperature and pH on the catalytic activity of the immobilized β-galactosidase from Kluyveromyces lactis. Biochem Eng J 9:33–40

    Article  Google Scholar 

  • Zokaee F, Kaghazchi T, Zare A, Soleimani M (2002) Isomerization of lactose to lactulose-study and comparison of three catalytic systems. Process Biochem 37:629–635

    Article  Google Scholar 

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Acknowledgements

This study was supported by a grant of the 21C Frontier Project for Microbial Genomics, Ministry of Science and Technology, Republic of Korea.

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  1. Department of Bioscience and Biotechnology, Sejong University, 98 Gunja-Dong Gwangjin-Gu, 143–747, Seoul , South Korea

    Y.-J. Lee, C. S. Kim & D.-K. Oh

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  1. Y.-J. Lee
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  2. C. S. Kim
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  3. D.-K. Oh
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Correspondence to D.-K. Oh.

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Lee, YJ., Kim, C.S. & Oh, DK. Lactulose production by β-galactosidase in permeabilized cells of Kluyveromyces lactis . Appl Microbiol Biotechnol 64, 787–793 (2004). https://doi.org/10.1007/s00253-003-1506-1

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  • DOI: https://doi.org/10.1007/s00253-003-1506-1

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