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European Food Research and Technology

, Volume 236, Issue 5, pp 817–826 | Cite as

Lactobacillus plantarum 70810 from Chinese paocai as a potential source of β-galactosidase for prebiotic galactooligosaccharides synthesis

  • Hongzhi Zhang
  • Wei Li
  • Xin Rui
  • Xingmin Sun
  • Mingsheng Dong
Original Paper

Abstract

A novel food-grade strain Lactobacillus plantarum 70810 producing β-galactosidase with high transgalactosylation activity was isolated from Chinese paocai. The galactooligosaccharides (GOS) were synthesized by using this enzyme with a maximum yield of 44.3 % (w/w) from 400 g/L lactose at 45 °C for 10 h. The β-galactosidase from this strain was purified to homogeneity by ammonium sulfate precipitation, anion exchange chromatography and gel filtration chromatography. It was a heterodimer arrangement of approximately 105 kDa composed of two subunits of 35 and 72 kDa. The optimal pH of the purified β-galactosidase was 8.0 for both o-nitrophenyl-β-d-galactopyranoside (oNPG) and lactose hydrolysis, and optimal temperature was 60 °C and 55 °C, respectively. Its K m and V max values for oNPG and lactose were 0.89 ± 0.05 mM, 194 ± 3.0 μmoL/min/mg protein, and 9.88 ± 0.16 mM, 15.88 ± 0.21 μmoL/min/mg protein, respectively. This enzyme was slightly inhibited by the hydrolysis products, that is, glucose and galactose. Since the β-galactosidase from L. plantarum 70810 exhibited higher transgalactosylation activity, strong affinity for lactose and low end-product inhibition, it was suggested to be a potential candidate for the synthesis of prebiotic GOS.

Keywords

Lactobacillus plantarum 70810 β-Galactosidase Chinese paocai Transgalactosylation Galactooligosaccharides 

Notes

Acknowledgments

This work was cofinanced by High-Tech Research and Development Program of China (No. 2011AA100903), National Natural Science Foundation of China (No. 31201422), Research Fund for the Doctoral Program of Higher Education of China, State Education Ministry (No. 20110097120028), Natural Science Foundation of Jiangsu Province (No. BK2011651), Youth Science and Technology Innovation Fund of Nanjing Agricultural Univerisity (No. KJ2010018) and was also supported by the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Conflict of interest

None.

Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.

References

  1. 1.
    Gibson GR, Roberfroid MB (1995) Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr 125:1401–1412Google Scholar
  2. 2.
    Bouhnik Y, Raskine L, Simoneau G, Vicaut E, Neut C, Flourié B, Brouns F, Bornet FR (2004) The capacity of nondigestible carbohydrates to stimulate fecal bifidobacteria in healthy humans: a double-blind, randomized, placebo-controlled, parallel-group, dose-response relation study. Am J Clin Nutr 80:1658–1664Google Scholar
  3. 3.
    Torres DPM, Goncalves MPF, Teixeira JA, Rodrigues LR (2010) Galacto-oligosaccharides: production, properties, applications, and significance as prebiotics. Compr Rev Food Sci F 9:438–454CrossRefGoogle Scholar
  4. 4.
    Crittenden RG, Playne MJ (1996) Production, properties and applications of food-grade oligosaccharides. Trends Food Sci Technol 7:353–361CrossRefGoogle Scholar
  5. 5.
    Gosling A, Stevens GW, Barber AR, Kentish SE, Gras SL (2010) Recent advances refining galactooligosaccharide production from lactose. Food Chem 121:307–318CrossRefGoogle Scholar
  6. 6.
    Park A, Oh DK (2010) Galacto-oligosaccharide production using microbial β-galactosidase: current state and perspectives. Appl Microbiol Biotechnol 85:1279–1286CrossRefGoogle Scholar
  7. 7.
    Li W, Sun Y, Ye H, Zeng XX (2010) Synthesis of oligosaccharides with lactose and N-acetylglucosamine as substrates by using β-d-galactosidase from Bacillus circulans. Eur Food Res Technol 231:55–63CrossRefGoogle Scholar
  8. 8.
    Panesar PS, Panesar R, Singh RS, Kennedy JF, Kumar H (2006) Microbial production, immobilization and applications of β-d-galactosidase. J Chem Technol Biotechnol 81:530–543CrossRefGoogle Scholar
  9. 9.
    Iqbal S, Nguyen TH, Nguyen TT, Maischberger T, Haltrich D (2010) β-Galactosidase from Lactobacillus plantarum WCFS1: biochemical characterization and formation of prebiotic galacto-oligosaccharides. Carbohydr Res 345:1408–1416CrossRefGoogle Scholar
  10. 10.
    Splechtna B, Nguyen TH, Zehetner R, Lettner HP, Lorenz W, Haltrich D (2007) Process development for the production of prebiotic galacto-oligosaccharides from lactose using β-galactosidase from Lactobacillus sp. Biotechnol J 2:480–485CrossRefGoogle Scholar
  11. 11.
    Tzortzis G, Goulas AK, Gibson GR (2005) Synthesis of prebiotic galactooligosaccharides using whole cells of a novel strain, Bifidobacterium bifidum NCIMB 41171. Appl Microbiol Biotechnol 68:412–416CrossRefGoogle Scholar
  12. 12.
    Lim JS, Lee JH, Kang SW, Park SW, Kim SW (2007) Studies on production and physical properties of neo-FOS produced by co-immobilized Penicillium citrinum and neo-fructosyltransferase. Eur Food Res Technol 225:457–462CrossRefGoogle Scholar
  13. 13.
    Rabiu BA, Jay AJ, Gibson GR, Rastall RA (2001) Synthesis and fermentation properties of novel galacto-oligosaccharides by β-galactosidase from Bifidobacterium species. Appl Environ Microbiol 67:2526–2530CrossRefGoogle Scholar
  14. 14.
    Iqbal S, Nguyen TH, Nguyen HA, Nguyen TT, Maischberger T, Kittl R, Haltrich D (2011) Characterization of a heterodimeric GH2 β-galactosidase from Lactobacillus sakei Lb790 and formation of prebiotic galacto-oligosaccharides. J Agric Food Chem 59:3803–3811CrossRefGoogle Scholar
  15. 15.
    Rastall RA, Maitin V (2002) Prebiotics and synbiotics: towards the next generation. Curr Opin Biotechnol 13:490–496CrossRefGoogle Scholar
  16. 16.
    Gänzle MG (2012) Enzymatic synthesis of galacto-oligosaccharides and other lactose derivatives (hetero-oligosaccharides) from lactose. Int Dairy J 22:116–122CrossRefGoogle Scholar
  17. 17.
    Cardelle-Cobas A, Corzo N, Martínez-Villaluenga C, Olano A, Villamiel M (2011) Effect of reaction conditions on lactulose-derived trisaccharides obtained by transgalactosylation with β-galactosidase of Kluyveromyces lactis. Eur Food Res Technol 233:89–94CrossRefGoogle Scholar
  18. 18.
    Feng MQ, Chen XH, Li CC, Nurgul R, Dong MS (2012) Isolation and identification of an exopolysaccharide-producing lactic acid bacterium strain from Chinese paocai and biosorption of Pb(II) by its exopolysaccharide. J Food Sci 77:T111–T117CrossRefGoogle Scholar
  19. 19.
    Cardelle-Cobas A, Martínez-Villaluenga C, Sanz ML, Montilla A (2009) Gas chromatographic–mass spectrometric analysis of galactosyl derivatives obtained by the action of two different β-galactosidases. Food Chem 114:1099–1105CrossRefGoogle Scholar
  20. 20.
    Li W, Xiang XL, Tang S, Hu B, Tian L, Sun Y, Ye H, Zeng XX (2009) Effective enzymatic synthesis of lactosucrose and its analogues by β-d-galactosidase from Bacillus circulans. J Agric Food Chem 57:3927–3933CrossRefGoogle Scholar
  21. 21.
    Leary NO, Pembroke A, Duggan PF (1992) Improving accuracy of glucose oxidase procedure for glucose determinations on discrete analyzers. Clin Chem 38:298–302Google Scholar
  22. 22.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  23. 23.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277:680–685CrossRefGoogle Scholar
  24. 24.
    Nguyen TH, Splechtna B, Steinböck M, Kneifel W, Lettner HP, Kulbe KD, Haltrich D (2006) Purification and characterization of two novel β-galactosidases from Lactobacillus reuteri. J Agric Food Chem 54:4989–4998CrossRefGoogle Scholar
  25. 25.
    Hung MN, Lee BH (2002) Purification and characterization of a recombinant β-galactosidase with transgalactosylation activity from Bifidobacterium infantis HL96. Appl Microbiol Biotechnol 58:439–445CrossRefGoogle Scholar
  26. 26.
    Hsu CA, Lee SL, Chou CC (2007) Enzymatic production of galactooligosaccharides by β-galactosidase from Bifidobacterium longum BCRC 15708. J Agric Food Chem 55:2225–2230Google Scholar
  27. 27.
    Nguyen TH, Splechtna B, Krasteva S, Kneifel W, Kulbe KD, Divne C, Haltrich D (2007) Characterization and molecular cloning of a heterodimeric β-galactosidase from the probiotic strain Lactobacillus acidophilus R22. FEMS Microbiol Lett 269:136–144CrossRefGoogle Scholar
  28. 28.
    Adams RM, Yoast S, Mainzer SE, Moon K, Palombella AL, Estell DA, Power SD, Schmidt BF (1994) Characterization of two cold-sensitive mutants of the v-galactosidase from Lactobacillus delbrückii subsp. Bulgaricus. J Biol Chem 269:5666–5672Google Scholar
  29. 29.
    Nadder de Macias ME, Manca de Nadra MC, Strasser de Saad AM, Pesce de Ruiz Holgado AA, Oliver G (1983) Isolation and properties of β-galactosidase of a strain of Lactobacillus helveticus isolated from natural whey starter. J Appl Biochem 5:275–281Google Scholar
  30. 30.
    Tello-Solis SR, Jimenez-Guzman J, Sarabia-Leos C, Gomez-Ruiz L, Cruz-Guerrero AE, Rodriguez-Serrano G, García-Garibay M (2005) Determination of the secondary structure of Kluyveromyces lactis β-galactosidase by circular dichroism and its structure–activity relationship as a function of the pH. J Agric Food Chem 53:10200–10204CrossRefGoogle Scholar
  31. 31.
    Vetere A, Paoletti S (1998) Separation and characterization of three β-galactosidases from Bacillus circulans. Biochim Biophys Acta 1380:223–231CrossRefGoogle Scholar
  32. 32.
    Maischberger T, Leitner E, Nitisinprasert S, Juajun O, Yamabhai M, Nguyen TH, Haltrich D (2010) β-Galactosidase from Lactobacillus pentosus: purification, characterization and formation of galacto-oligosaccharides. Biotechnol J 5:838–847CrossRefGoogle Scholar
  33. 33.
    De Roos A (2004) Industrial enzymes: enzymes in dairy applications. In: Aehle W (ed) Enzymes in industry, 2nd edn. Wilsey-VCH, WeinheimGoogle Scholar
  34. 34.
    Kim CS, Ji ES, Oh DK (2003) Expression and characterization of Kluyveromyces lactis β-galactosidase in Escherichia coli. Biotechnol Lett 25:1769–1774CrossRefGoogle Scholar
  35. 35.
    Hsu CA, Yu RC, Chou CC (2006) Purification and characterization of a sodium-stimulated β-galactosidase from Bifidobacterium longum CCRC15708. World J Microbiol Biotechnol 22:355–361CrossRefGoogle Scholar
  36. 36.
    Splechtna B, Nguyen TH, Steinböck M, Kulbe KD, Lorenz W, Haltrich D (2006) Production of prebiotic galacto-oligosaccharides from lactose using β-galactosidase from Lactobacillus reuteri. J Agric Food Chem 54:4999–5006CrossRefGoogle Scholar
  37. 37.
    Rhimi M, Boisson A, Dejob M, Boudebouze S, Maguin E, Hasera R, Aghajaria N (2010) Efficient bioconversion of lactose in milk and whey: immobilization and biochemical characterization of a β-galactosidase from the dairy Streptococcus thermophilus LMD9 strain. Res Microbiol 161:515–525CrossRefGoogle Scholar
  38. 38.
    Yi SH, Alli I, Park KH, Lee BH (2011) Overexpression and characterization of a novel transgalactosylic and hydrolytic β-galactosidase from a human isolate Bifidobacterium breve B24. New Biotechnol 28:806–813CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Hongzhi Zhang
    • 1
  • Wei Li
    • 1
  • Xin Rui
    • 1
  • Xingmin Sun
    • 2
  • Mingsheng Dong
    • 1
  1. 1.College of Food Science and TechnologyNanjing Agricultural UniversityNanjingPeople’s Republic of China
  2. 2.Department of Biomedical SciencesTufts Cummings School of Veterinary MedicineNorth GraftonUSA

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