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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제22권 제12호
- 발행연도
- 2012.1
- 수록면
- 1,705 - 1,713 (9page)
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초록· 키워드
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A gene encoding β-glucosidase was cloned from Weissella cibaria 37, an isolate from human feces. Sequence analysis showed that the gene could encode a protein of 415 amino acids in length, and the translated amino acid sequence showed homology (34-31%) with glycosyl hydrolase family 1 β-glucosidases. The gene was overexpressed in E.
coli BL21(DE3) using pET26b(+) and a 50 kDa protein was overproduced, which matched well with the calculated size of the enzyme, 49,950.87 Da. Recombinant β-glucosidase was purified by using a his-tag affinity column. The purified β-glucosidase had an optimum pH and a temperature of 5.5 and 45oC, respectively. Among the metal ions (5mM concentration), Ca2+ slightly increased the activity (108.2%) whereas Cu2+ (46.1%) and Zn2+ (56.7%)reduced the activity. Among the enzyme inhibitors (1 mM concentration), SDS was the strongest inhibitor (16.9%),followed by pepstatin A (45.2%). The Km and Vmax values of purified enzyme were 4.04 mM and 0.92 μmol/min,respectively, when assayed using pNPG (p-nitrophenyl-β-D-glucopyranoside) as the substrate. The enzyme liberated reducing sugars from carboxymethyl cellulose (CMC).
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참고문헌 (22)
참고문헌 신청
Bauer, M. W. / 1996 / Comparison of a β-glucosidase and a β-mannosidasefrom the hyperthermophilic Archaeon Pyrococcus furiosus / J.Biol. Chem 271 : 23749 ~ 23755
Belancic, A. / 2003 / Beta-glucosidase from the grape native yeast Debrayomyces; purification, characterization, and its effect on monoterpene content of a muscat grape juice
Benoliel, B. / 2010 / Expression of a glucose-tolerant β-glucosidase from Humicola grisea var. thermoidea in Saccharomyces cerevisiae / Appl. Biochem. Biotechnol 160 : 2036
Bhatia, Y. / 2002 / Microbial β- glucosidases: Cloning, properties, and applications / Crit. Rev. Biotechnol 22 : 375 ~ 407
Bradford, M. M. / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 7
Belancic, A. / 2003 / Beta-glucosidase from the grape native yeast Debrayomyces; purification, characterization, and its effect on monoterpene content of a muscat grape juice
Benoliel, B. / 2010 / Expression of a glucose-tolerant β-glucosidase from Humicola grisea var. thermoidea in Saccharomyces cerevisiae / Appl. Biochem. Biotechnol 160 : 2036
Bhatia, Y. / 2002 / Microbial β- glucosidases: Cloning, properties, and applications / Crit. Rev. Biotechnol 22 : 375 ~ 407
Bradford, M. M. / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 7
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