Skip to main content

Advertisement

SpringerLink
Log in

Purification and characterization of hydrolytic and transgalactosyl α-galactosidase from Lactobacillus helveticus ATCC 10797

  • Original Paper
  • Published:
European Food Research and Technology Aims and scope Submit manuscript

Abstract

α-Galactosidase purified from Lactobacillus helveticus ATCC 10797 by fast performance liquid chromatography system using ion exchange and gel-filtration columns showed the K m of 3.83 mM and V max of 416.44 µmol/min/mg protein calculated from the substrate p-nitrophenyl-α-d-galactopyranoside. The molecular mass was 188 kDa by gel-filtration, but 90 kDa by SDS-PAGE, indicating a homodimer. The optimum temperature was 37 °C, and the optimum pH was at 6 with an acceptable stability between pH 4 and 8. This enzyme was activated by 10 mM monovalent ions such as K+, NH4 +, Li+, and CS+, while the activity was inhibited by divalent ions such as Cu2+, Zn2+, and Fe2+. Melibiose was hydrolyzed to glucose and galactose, raffinose to galactose and sucrose, while stachyose to galactose and sucrose. A novel source of α-galactosidase from L. helveticus possessing both hydrolytic activity to eliminate flatulence sugars and transgalactosylation activities to synthesize galacto-oligosaccharides is identified and characterized.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Crittenden RG, Playne MJ (1996) Production, properties, and applications of food-grade oligosaccharides. Trends Food Sci Technol 7(11):353–361

    Article  CAS  Google Scholar 

  2. Kang HC, Lee SH (2001) Characteristics of α-galactosidase associated with grape flesh. Phytochem 58:213–219

    Article  CAS  Google Scholar 

  3. Baek YJ, Lee BH (2009) Probiotics and prebiotics as bioactive components in dairy products. In: Park YW (ed) Bioactive components in milk and dairy products. Wiley-Blackwell Publ, New York, p 449

    Google Scholar 

  4. Machaiah JP, Pednekar MD, Thomas P (1999) Reduction in flatulence factors in mung beans (Vigna radiata) using low-dose gamma -irradiation. J Sci Food Agric 79:648–652

    Article  CAS  Google Scholar 

  5. Yamaguishi CT, Sanada CT, Gouvea PM, Pandey A, Patricia M, Woiciechowski AL, Parada JL, Soccol CR (2009) Biotechnological process for producing black bean slurry without stachyose. Food Res Intern 42:425–429

    Article  CAS  Google Scholar 

  6. LeBlanc JG, Ledue-Clier F, Bensaada M, de Giori GS, Guerekobaya T, Sesma F, Juillard V, Rabot S, Piard JC (2008) Ability of Lactobacillus fermentum to overcome host alpha- galactosidase deficiency, as evidenced by reduction of hydrogen excretion in rats consuming soya alpha-galacto-oligosaccharides. BMC Microbiol 8:22–26

    Article  Google Scholar 

  7. Di Stefano M, Miceli E, Armellini E, Missanelli A, Corazza GR (2004) Probiotics and functional abdominal bloating. J Clin Gastroenterol 38:S102–S103

    Article  Google Scholar 

  8. Zhao H, Lu LL, Xiao M, Wang Q, Yu L (2008) Cloning and characterization of a novel α- galactosidase from Bifidobacterium breve 203 capable of synthesizing Gal-α-1,4 linkage. FEMS Microbiol Lett 285:278–283

    Article  CAS  Google Scholar 

  9. Katrolia P, Huiyong J, Qiaojuan Y, Song S, Jiang Z, Xu H (2012) Characterization of a protease-resistant α-galactosidase from the thermophilic fungus Rhizomucor miehei and its application in removal of raffinose family oligosaccharides. Bioresour Technol 110:578–586

    Article  CAS  Google Scholar 

  10. Lee BH, Hache S, Simard RE (1986) A rapid method for differentiation of dairy lactic acid bacteria by enzyme systems. J Ind Microbiol 1:209–217

    Article  CAS  Google Scholar 

  11. Xiaoli Liu, Champagne CP, Lee BH, Boye JI, Casgrain M (2013) Thermostability of probiotics and their α-galactosidases, and the potential for bean processing. Biotech Res Intern. Article ID 472723, 21 pages, http://dx.doi.org/10.1155/2014/472723

  12. Scalabrini P, Rossi M, Spettoli P, Matteuzzi D (1998) Characterization of strains for use in soymilk fermentation. Intern J Food Microbiol 39:213–219

    Article  CAS  Google Scholar 

  13. Garro MS, Valdez GF, Oliver G, Giori GS (1996) Purification of α-galactosidase from Lactobacillus fermentum. J Bacteriol 45:103–109

    CAS  Google Scholar 

  14. Sugimoto H, Van Buren JM (1970) Removal of oligosaccharides from soymilk by an enzyme from Aspergillus saitoi. J Food Sci 35:655–660

    Article  CAS  Google Scholar 

  15. McGhee JE, Silman R, Bagley EB (1978) Production of α-galactosidase from Aspergillus awamori: properties and action on para-nitrophenyl-α-d-galactopyranoside and galacto-oligosaccharides of soy milk. J Am Oil Chem 55:244–247

    Article  CAS  Google Scholar 

  16. Church FC, Meyers SP, Srinivasan VR (1980) Isolation and characterization of α-galactosidase from Pichia guilliermondii. In: Underkofler LA, Wulf ML (eds) Developments in industrial microbiology, vol 21. Society for industrial microbiology, Arlington, pp 339–348

    Google Scholar 

  17. Kawamura S, Kasai T, Tanusi S (1976) Purification and properties of alpha- galactosidase from Escherichia coli subsp. communior IAM 1272. Agric Biol Chem 40:641–648

    Article  CAS  Google Scholar 

  18. Kusumoto K, Shirahata S, Kamei Y (2000) Purification and characterization of alpha-d- galactosidase produced by ADG cell line established from abalon digestive gland. Cytotechnology 33:47–52

    Article  CAS  Google Scholar 

  19. Aihara K, Kajimoto O, Hirata H (2005) Effect of powdered fermented milk with Lactobacillus helveticus on subjects with high-normal blood pressure or mild hypertension. J Amer Coll Nutr 24:257–265

    Article  Google Scholar 

  20. Li YT, Shetlar MR (1964) Occurrence of α-galactosidase in higher fungi; isolation of α-galactosidase from Calvatia cyathiformis. Arch Biochem Biophys 108:523–530

    Article  CAS  Google Scholar 

  21. Suzoki H, Li SC, Li YT (1970) α-Galactosidase from Moteriella vinacea. Crystallization and properties. J Biol Chem 245:781–786

    Google Scholar 

  22. Kim GB, Lee BH (2008) Genetic analysis of a bile salt hydrolase (bsh) in Bifidobacterium animalis subsp.lactis KL612. J Appl Microbiol 105:778–789

    Article  CAS  Google Scholar 

  23. Fortier LF, Tourdot-Marechal R, Davies C, Lee BH, Guzzo J (2003) Induction of Oenococcus oeni H+-ATPase activity and mRNA transcription under acidic condition of growth. FEMS Microbiol Lett 222:165–169

    Article  CAS  Google Scholar 

  24. Dey PM, Pridham JB (1972) Biochemistry of α-galactosidase. Adv Enzymol Relat Areas Mol Biol 36:91–130

    CAS  Google Scholar 

  25. Mital BK, Steinkraus KH, Naylor HB (1975) Growth of lactic acid bacteria in soymilk. J Food Sci 39:1018–1022

    Article  Google Scholar 

  26. Wong HC, Hu CA, Yeh HL, Su W, Lu HC, Lin CF (1986) Production, purification, and characterization of α-galactosidase from Monascus pilosus. Appl Environ Microbiol 52:1147–1152

    CAS  Google Scholar 

  27. Grossmann GA, Terra WR (2001) Alpha-galactosidases from the larval midgut of Tenebrio molitor (coleoptera) and Spodoptera frugiperda (lepidoptera). Comp Biochem Physiol 128:109–122

    Article  CAS  Google Scholar 

  28. King MR, White BA, Blaschek HP, Chassy BM, Mackie RI, Cann IKO (2002) Purification and characterization of a thermostable α-galactosidase from Thermobacterium polysaccharolyticum. J Agric Food Chem 50:5672–5682

    Article  Google Scholar 

  29. Saulnier DM, Kolida S, Gibson GR (2009) Microbiology of the human intestinal tract and approaches for its dietary modulation. Curr Pharm Design 15:1403–1414

    Article  CAS  Google Scholar 

  30. Dey PM, Naik S, Pridham JB (1986) Properties of alpha-galactosidase II2 from Vicia faba seeds. Planta 167:114–118

    Article  CAS  Google Scholar 

  31. Zapater IG, Ullah AHJ, Wodzinsky RJ (1990) Extracellular alpha-galactosidase (E.C. 3.2.1.22) from Aspergillus ficuum NRRL 3135 purification and characterization. Prep Biochem Biotechnol 20:263–296

    CAS  Google Scholar 

  32. Cavazzoni V, Adami A, Craveri R (1987) Alpha-galactosidase from yeast Candida javanica. Appl Microbiol Biotechnol 26:555–559

    Article  CAS  Google Scholar 

Download references

Conflict of interest

No conflict of Interest exists in this study.

Compliance with ethics requirements

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

Author information

Authors and Affiliations

  1. Department of Food Science and Agricultural Chemistry, McGill University, Ste-Anne-de Bellevue, QC, H9X 3V9, Canada

    Sharifa Kandari & Byong H. Lee

  2. Life Sciences, NRC Human Health Therapeutics Portfolio, National Research Council of Canada, 6100 Royalmount Ave., Montreal, QC, H4P 2R2, Canada

    Young J. Choi

  3. School of Biotechnology, Jiangnan University, Wuxi, 214122, China

    Byong H. Lee

Authors
  1. Sharifa Kandari
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Young J. Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Byong H. Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Byong H. Lee.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kandari, S., Choi, Y.J. & Lee, B.H. Purification and characterization of hydrolytic and transgalactosyl α-galactosidase from Lactobacillus helveticus ATCC 10797. Eur Food Res Technol 239, 877–884 (2014). https://doi.org/10.1007/s00217-014-2284-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00217-014-2284-y

Keywords

Search

Navigation