Advertisement

Food Science and Biotechnology

, Volume 22, Issue 1, pp 63–70 | Cite as

Some characteristics and isolation of novel thermostable β-galactosidase from Thermus oshimai DSM 12092

  • Yüksel GezginEmail author
  • Bahattin Tanyolac
  • Rengin Eltem
Research Article

Abstract

The β-galactosidases belong to the class of hydrolytic enzymes and have been used as lactose hyrolysis. The enzyme is used in reducing lactose milk production, an outstanding industrial product used by a large lactoseintolerant population. This is the first detailed report of some characteristics of β-galactosidase and the gene encoding β-galactosidase in Thermus oshimai DSM 12092. The growth rate (μ, 1/h), and the doubling time (tD, h) for T. oshimai grown both in shaking flasks and in a bioreactor were determined. The optimal temperature and pH for β-galactosidase were determined as 75°C and 7.4, respectively. This enzyme was thermostable and was retained by more than 70% at 90°C for 3 h. The β-galactosidase from T. oshimai DSM 12092 was more stable in basic pH and Zn2+ was the most effective divalent cation. Also, 2 steps of purification consisting of ammonium sulfate precipitation and gel filtration were employed and purified 32-fold.

Keywords

β-galactosidase T. oshimai DSM 12092 thermostability galactooligosaccarides synthesis 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Maciunska J, Czyz B, Synowiecki J. Isolation and some properties of β-galactosidase from the thermophilic bacterium Thermus thermophilus. Food Chem. 63: 441–445 (1998)CrossRefGoogle Scholar
  2. 2.
    Chen W, Chen H, Xia Y, Yang J, Zhao J, Tian F, Zhang HP, Zhang H. Immobilization of recombinant thermostable β-galactosidase from Bacillus stearothermophilus for lactose hydrolysis in milk. J. Dairy Sci. 92: 491–498 (2009)CrossRefGoogle Scholar
  3. 3.
    Berger JL, Lee BH, Lacroix C. Oligosaccharide synthesis by free and immobilized β-galactosidases from Thermus aquaticus YT-1. Biotechnol. Lett. 17: 1077–1080 (1995)CrossRefGoogle Scholar
  4. 4.
    Grosova Z, Rosenberg M, Rebroš M. Perspectives and applications of immobilised β-galactosidase in food industry — A review. Czech J. Food Sci. 26: 1–14 (2008)Google Scholar
  5. 5.
    Panesar PS, Panesar R, Singh RS, Kennedy JF, Kumar H. Microbial production, immobilization, and applications of β-D-galactosidase. J. Chem. Technol. Biot. 81: 530–543 (2006)CrossRefGoogle Scholar
  6. 6.
    Park, AR, Oh DK. Galacto-oligosaccharide production using microbial β-galactosidase: Current state and perspectives. Appl. Microbiol. Biot. 85: 1279–1286 (2010)CrossRefGoogle Scholar
  7. 7.
    Gosling A, Stevens GW, Barber AR, Kentish SE, Gras SL. Recent advances refining galactooligosaccharide production from lactose. Food Chem. 121: 307–318 (2010)CrossRefGoogle Scholar
  8. 8.
    Pisani FM, Rella R, Raia CA, Rozzo C, Nucci R, Gambacorta A, Rosa MD, Rossi M. Thermostable β-galactosidase from the archaebacterium Sulfolobus solfataricus purification and properties. Eur. J. Biochem. 187: 321–328 (1990)CrossRefGoogle Scholar
  9. 9.
    Chen W, Chen H, Xia Y, Zhao J, Tian F, Zhang H. Production, purification, and characterization of a potential thermostable galactosidase for milk lactose hydrolysis from Bacillus stearothermophilus. J. Dairy Sci. 91: 1751–1758 (2008)CrossRefGoogle Scholar
  10. 10.
    Ohtsu N, Motoshıma H, Goto K, Tsukasaki F, Matsuzawa H. Thermostable β-galactosidase from an extreme thermophile, Thermus sp. A4: Enzyme purification and characterization, and gene cloning and sequencing. Biosci. Biotech. Bioch. 62: 1539–1545 (1998)CrossRefGoogle Scholar
  11. 11.
    Koyama Y, Hoshino T, Tomizuka N, Furukawa K. Genetic transformation of the extreme thermophile Thermus thermophilus and of other Thermus spp. J. Bacteriol. 166: 338–340 (1986)Google Scholar
  12. 12.
    Kim CS, Ji ES, Oh DK. Characterization of a thermostable recombinant β-galactosidase from Thermotoga maritima. J. Appl. Microbiol. 97: 1006–1014 (2004)CrossRefGoogle Scholar
  13. 13.
    Vian A, Carrascosa AV, García JL, Cortés E. Structure of the β-galactosidase gene from Thermus sp. strain T2: Expression in Escherichia coli and purification in a single step of an active fusion protein. Appl. Environ. Microb. 64: 2187–2191 (1998)Google Scholar
  14. 14.
    Nam ES, Choi JW, Lim JH, Hwang SK, Jung HJ, Kang SK, Cho KK, Choi YJ, Ahn JK. β-Galactosidase gene of Thermus thermophilus KNOUC112 isolated from hot springs of a volcanic area in New Zealand: Identification of the bacteria, cloning, and expression of the gene in Escherichia coli. Asian Austral. J. Anim. Sci. 17: 1591–1598 (2004)Google Scholar
  15. 15.
    Fridjonsson O, Watzlawick H, Gehweiler A, Rohrhirsch T, Mattes R. Cloning of the gene encoding a novel thermostable α-galactosidase from Thermus brockianus ITI360. Appl. Environ. Microb. 65: 3955–3963 (1999)Google Scholar
  16. 16.
    Kang SK, Cho KK, Ahn JK, Bok JD, Kang SH, Woo JH, Lee HG, You SK, Choi YJ. Three forms of thermostable lactose-hydrolase from Thermus sp. IB-21: Cloning, expression, and enzyme characterization. J. Biotechnol. 116: 337–346 (2005)CrossRefGoogle Scholar
  17. 17.
    Miller JH. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory Press, New York, NY, USA. p. 468 (1972)Google Scholar
  18. 18.
    Ornelas AP, Silveira WB, Sampaio FC, Passos FML. The activity of β-galactosidase and lactose metabolism in Kluyveromyces lactis cultured in cheese whey as a function of growth rate. J. Appl. Microbiol. 104: 1008–1013 (2007)CrossRefGoogle Scholar
  19. 19.
    Bradford MM. A rapid and sensitive for the quantitation of microgram quantitites of protein utilizing the principle of proteindye binding. Anal. Biochem. 72: 248–254 (1976)CrossRefGoogle Scholar
  20. 20.
    Hsu CA, Lee SL, Chou CC. Enzymatic production of galactooligosaccharides by β-galactosidase from Bifidobacterium longum BCRC 15708. J. Agr. Food. Chem. 55: 2225–2230 (2007)CrossRefGoogle Scholar
  21. 21.
    Demirtas MU, Kolhatkar A, Kýlbane JJ. Effect of aeration and agitation on growth rate of Thermus thermophilus in batch mode. J. Biosci. Bioeng. 95: 113–117 (2003)Google Scholar
  22. 22.
    Ranzi BM, Porro D, Compagno C, Martegani E. Protein and cell volume distributions during the production of β-galactosidase in batch cultures of Kluyveromyces lactis. J. Biotechnol. 5: 227–231 (1987)CrossRefGoogle Scholar
  23. 23.
    Dickson RC, Markin JS. Physiological studies of β-galactosidase induction in Kluyveromyces lactis. J. Bacteriol. 142: 777–785 (1980)Google Scholar
  24. 24.
    Barberis S, Segovia R. Maximum volumetric production of β-galactosidase by Kluyveromyces fragilis in fed-batch culture with automatic control. J. Chem. Technol. Biot. 77: 706–710 (2002)CrossRefGoogle Scholar
  25. 25.
    Pinheiro R, Belo I, Mota M. Growth and β-galactosidase activity in cultures of Kluyveromyces marxianus under increased air pressure. Lett. Appl. Microbiol. 37: 438–442 (2003)CrossRefGoogle Scholar
  26. 26.
    Ladero M, Ferrero R, Vian A, Santos A, Garcia-Ochoa F. Kinetic modelling of the thermal and pH inactivation of a termostable β-galactosidases from Thermus sp. strain T2. Enzyme Microb. Tech. 37: 505–513 (2005)CrossRefGoogle Scholar
  27. 27.
    Ladero M, Santos A, Garcia JL, Carrascosa AV, Pessela BCC, Garcia-Ochoa F. Studies on the activity and the stability of β-galactosidases from Thermus sp. strain T2 and from Kluyveromyces fragilis. Enzyme Microb. Tech. 30: 392–405 (2002)CrossRefGoogle Scholar
  28. 28.
    Cowan, DA, Daniel RM, Morgan HW. Some properties of β-galactosidase from an extremely thermophilic bacterium. Biotechnol. Bioeng. 26: 1141–1145 (1984)CrossRefGoogle Scholar
  29. 29.
    Koyama Y, Okamoto S, Furukawa K. Cloning of α- and β-galactosidase genes from an extreme thermophile, Thermus strain T2, and their expression in Thermus thermophilus HB27. Appl. Microbiol. Biot. 56: 2251–2254 (1990)Google Scholar

Copyright information

© The Korean Society of Food Science and Technology and Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Yüksel Gezgin
    • 1
    Email author
  • Bahattin Tanyolac
    • 1
  • Rengin Eltem
    • 1
  1. 1.Department of Bioengineering, Faculty of EngineeringEge UniversityIzmir-BornovaTurkey

Personalised recommendations