지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2013.4
- 수록면
- 97 - 111 (15page)
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초록· 키워드
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α-Glucosidases (HBGases) catalyze the non-reducing end of a substrate to liberate α-D-glucose, and are classified into the three types of HBGase I, II and III according to their substrate specificity. The full length ORF cDNA (1,734 bp) and the deduced encoded amino acid sequence (577 residues) of the HBGase I from Apis cerana indica (AciHBGase I) was obtained by RT-PCR. The deduced amino acid sequence showed 99% and 80% sequence identity to the HBGase I from the closely related honey bees, A. cerana japonica and A. mellifera, respectively. After cloning in the pEcoli-Nterm-6xHN expression vector and transforming into E. coli Rosetta (DE3) cells the maximal glucosidase activity (assumed to be due to the expression of the recombinant (r)(His)<SUB>6</SUB>-AciHBGase I enzyme) was found in the cell lysate (0.054U/mL; equivalent to 2.16U/mL culture medium) after 1mM IPTG induction for 3 h. The partially purified r(His)<SUB>6</SUB>-AciHGase I enzyme displayed an optimal glucosidase activity, in terms of the hydrolysis of p-nitrophenyl α-D-glucoside, at pH 3.5 and 40°C, whilst it was stable (> 90% residual activity) at pH 3.5-5.5 (24 h at 4°C) and up to 45°C (15 min exposure at pH 5.5). The most preferable substrate was sucrose (K<SUB>m</SUB> value of 10.82±0.39mM).
목차
Abstract
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
LITERATURE CITED
참고문헌 (27)
참고문헌 신청
Bause, E. / 1986 / Purification and characterization of trimming glucosidase I from Saccharomyces cerevisae / FEBS Lett 206:208~212
Bradford, M. M. / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindin / Anal. Biochem 72:248~254
Cregg, J. M. / 2009 / Expression in the yeast Pichia pastoris / Methods Enzymol 463:169~189
Daly, R. / 2005 / Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production / J. Mol. Recognit 18:119~138
Dhanawansa, R. / 2002 / Overexpression, purification and partial characterization of Saccharomyces cerevisiae processing α-glucosidase I / Glycobiology 12:229~234
Bradford, M. M. / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindin / Anal. Biochem 72:248~254
Cregg, J. M. / 2009 / Expression in the yeast Pichia pastoris / Methods Enzymol 463:169~189
Daly, R. / 2005 / Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production / J. Mol. Recognit 18:119~138
Dhanawansa, R. / 2002 / Overexpression, purification and partial characterization of Saccharomyces cerevisiae processing α-glucosidase I / Glycobiology 12:229~234
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UCI(KEPA) : I410-ECN-0101-2014-520-002595708