DOI QR코드

DOI QR Code

청국장 유래 Bacillus licheniformis의 ${\beta}$-Galactosidase 특성

Properties of ${\beta}$-Galactosidase from Bacillus licheniformis Isolated from Cheongkookjang

  • 윤기홍 (우송대학교 식품생물과학과)
  • Yoon, Ki-Hong (Department of Food Science & Biotechnology, Woosong University)
  • 투고 : 2012.02.07
  • 심사 : 2012.03.10
  • 발행 : 2012.03.28

초록

가정에서 제조된 청국장으로부터 lactose를 glucose와 galactose로 가수분해하는 ${\beta}$-galactosidase의 생산균을 분리하였다. 분리균 YB-1105는 형태적 특성, 생화학적 성질 및 16S rRNA 유전자 염기서열에 근거하여 Bacillus licheniformis로 확인되었다. B. licheniformis YB-1105의 배양상등액과 균체파쇄액에서 모두 ${\beta}$-galactosidase 활성이 관찰되었으며 이들은 모두 pH 6.5와 $50^{\circ}C$의 반응조건에서 paranitrophenyl-${\beta}$-D-galactopyranoside의 가수분해 활성이 최대로 나타났다. 그러나 균체파쇄상등액에 비해 배양상등액의 ${\beta}$-galactosidase 활성은 산성 pH와 고온에서 크게 영향을 받았다. 한편 두 분획의 가수분해 활성은 낮은 농도의 galactose에 의해도 급격하게 저해되었으나, glucose와 mannose는 고농도에 의해서는 약하게 저해를 받는 것으로 확인되었다.

A bacterial strain was isolated from homemade Cheongkookjang as a producer of the ${\beta}$-galactosidase, capable of hydrolyzing lactose to liberate galactose and glucose residues. The isolate YB-1105 has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. ${\beta}$-Galactosidase activity was detected in both the culture supernatant and the cell extract of B. licheniformis YB-1105. The enzymes of both fractions demonstrated maximum activity for hydrolysis of para-nitrophenyl-${\beta}$-D-galactopyranoside (pNP-${\beta}Gal$) under identical reaction conditions of pH 6.5 and $50^{\circ}C$. However, ${\beta}$-galactosidase activity from the culture filtrate was affected more than that from the cell free extract at acidic pHs and high temperatures. The hydrolyzing activity of both ${\beta}$-galactosidases for pNP-${\beta}Gal$ was dramatically decreased by the addition of low concentrations of galactose, but was only marginally decreased by high concentrations of glucose or mannose.

키워드

참고문헌

  1. Anema, P. J. 1964. Purification and some properties of $\beta$-galactosidase of Bacillus subtilis. Biochim. Biophy. Acta. 89: 495-502.
  2. Batra, N., J. Singh, A. Joshi, and S. Bhatia. 2011. Applications of $\beta$-gal-III isozyme from Bacillus coagulans RCS3, in lactose hydrolysis. Int. J. Biol. Macromol. 49: 879-884.
  3. Batra, N., J. Singh, U. C. Banerjee, P. R. Patnaik, and R. C. Sobti. 2002. Production and characterization of a thermostable $\beta$-galactosidase from Bacillus coagulans RCS3. Biotechnol. Appl. Biochem. 36: 1-6.
  4. Chen, W., H. Chen, Y. Xia, J. Yang, J. Zhao, F. Tian, H. P. Zhang, and H. Zhang. 2009. Immobilization of recombinant thermostable $\beta$-galactosidase from Bacillus stearothermophilus for lactose hydrolysis in milk. J. Dairy Sci. 92: 491-498.
  5. Chen, W., H. Chen, Y. Xia, J. Zhao, F. Tian, and H. Zhang. 2008. Production, purification, and characterization of a potential thermostable galactosidase for milk lactose hydrolysis from Bacillus stearothermophilus. J. Dairy Sci. 91: 1751-1758.
  6. Goodman, R. E. and D. M. Pederson. 1976. $\beta$-Galactosidase from Bacillus stearothermophilus. Can. J. Microbiol. 22: 817-825.
  7. Hernaiz, M. J. and D. H. G Crout. 2000. A highly selective synthesis of N-acetyllactosamine catalyzed by immobilised $\beta$-galactosidase from Bacillus circulans. J. Mol. Catalysis B. 10: 403-408.
  8. Hirata, H., S. Negoro, and H. Okada. 1984. Molecular basis of isozyme formation of $\beta$-galactosidases in Bacillus stearothermophilus: isolation of two $\beta$-galactosidase genes, bgaA and bgaB. J. Bacteriol. 160: 9-14.
  9. Hong, S. W., J. Y. Kim, B. K. Lee, and K. S. Chung. 2006. The bacterial biological response modifier enriched Cheongkukjang fermentation. Kor. J. Food Sci. Technol. 38: 548-553.
  10. Ito, Y. and T. Sasaki. 1997. Cloning and characterization of the gene encoding a novel $\beta$-galactosidase from Bacillus circulans. Biosci. Biotechnol. Biochem. 61: 1270-1276.
  11. Juajun, O., T. H. Nguyen, T. Maischberger, S. Iqbal, D. Haltrich, and M. Yamabhai. 2011. Cloning, purification, and characterization of $\beta$-galactosidase from Bacillus licheniformis DSM 13. Appl. Microbiol. Biotechnol. 89: 645-654.
  12. Lee, G. H. and K. -H. Yoon. 2009. Sensory comparison of fermented soybeans (Cheongkookjang) inoculated with various Bacillus species by principal component analysis. J. Kor. Soc. Appl. Biol. Chem. 52: 726-730.
  13. Lee, K. S., C. -J. Kim, and K. -H. Yoon. 2003. Characterization of the $\beta$-galactosidase produced by Streptomyces sp. YB-10. Kor. J. Microbiol. Biotechnol. 31: 151-156.
  14. Li, Y., H. Wang, L. Lu, Z. Li, X. Xu, and M. Xiao. 2009. Purification and characterization of a novel $\beta$-galactosidase with transglycosylation activity from Bacillus megaterium 2- 37-4-1. Appl. Biochem. Biotechnol. 158: 192-199.
  15. Montero, E., J. Alonso, F. J. Canada, A. Fernandez- Mayoralas, and M. Martin-Lomas. 1997. Regioselectivity of the enzymatic transgalactosidation of D- and L-xylose catalysed by $\beta$-galactosidases. Carbohydr. Res. 305: 383-391.
  16. Pollard, H. B. and E. Jr. Steers. 1973. Bacillus megaterium, KM $\beta$-galactosidase: purification by affinity chromatography and characterization of the active species. Arch. Biochem. Biophys. 158: 650-661.
  17. Rahim, K. A. and B. H. Lee. 1991. Specificity, inhibitory studies, and oligosaccharide formation by $\beta$-galactosidase from psychrotrophic Bacillus subtilis KL88. J. Dairy Sci. 74: 1773-1778.
  18. Sanni, A. I., A. A. Onilude, and O. R. Ogundoye. 1997. Effect of bacterial galactosidase treatment on the nutritional status of soybean seeds and its milk derivative. Food/Nahrung 41: 18-21.
  19. Shaw, G. C., H. S. Kao, and C. Y. Chiou. 1998. Cloning, expression, and catabolite repression of a gene encoding $\beta$-galactosidase of Bacillus megaterium ATCC 14581. J. Bacteriol. 180: 4734-4738.
  20. Song, J., K. Abe, H. Imanaka, K. Imamura, M. Minoda, S. Yamaguchi, and K. Nakanishi. 2011. Causes of the production of multiple forms of $\beta$-galactosidase by Bacillus circulans. Biosci. Biotechnol. Biochem. 75: 268-278.
  21. Tran, L. S., L. Szabo, L. Fulop, L. Orosz, T. Sik, and A. Holczinger. 1998. Isolation of a $\beta$-galactosidase-encoding gene from Bacillus licheniformis: purification and characterization of the recombinant enzyme expressed in Escherichia coli. Curr. Microbiol. 37: 39-43.

피인용 문헌

  1. 된장에서 분리된 Bacillus licheniformis의 ${\beta}$-galactosidase 생산성과 효소특성 vol.42, pp.4, 2012, https://doi.org/10.4014/kjmb.1410.10004
  2. 전통장류 유래 Bacillus sp.를 이용한 콩 발효물의 품질 특성 vol.43, pp.5, 2014, https://doi.org/10.3746/jkfn.2014.43.5.756