Optimization of refolding conditions for the aklavinone 11-hydroxylase of Streptomyces peucetius overexpressed in Escherichia coli = 대장균에서 대량발현된 Streptomyces peucetius 유래 Aklavinone 11-hydroxylase 효소의 최적 가용화 조건

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Title
Optimization of refolding conditions for the aklavinone 11-hydroxylase of Streptomyces peucetius overexpressed in Escherichia coli = 대장균에서 대량발현된 Streptomyces peucetius 유래 Aklavinone 11-hydroxylase 효소의 최적 가용화 조건
Author(s)
Woo Keun Min; Young Soo Hong; Yong Kyung Choe; Jung Joon Lee; Soon Kwang Hong
Bibliographic Citation
Korean Journal of (Applied) Microbiology & Biotechnology, vol. 26, no. 4, pp. 365-368
Publication Year
1998
Abstract
The aklavinone 11-hydroxylase which was overexpressed using T7 promoter in E. coli could be detected in SDS-PAGE only in insoluble precipitate without any detectable enzyme activity. The insoluble enzyme was solubilized in 6M guanidine - HCl solution and their refolding ability was tested under various conditions. When the enzymatic activity was checked by the bioconversion experiment, stepwise dialysis against 6M, 3M, 1M guanidine · HCl and finally 100 mM potassium phosphate buffer of the solubilized protein gave the best bioconversion efficiency. The aklavinone 11-hydroxylase showed its enzymatic activity in the reaction buffer containing NADPH with vigorous shaking. The enzymatic activity was lost during partial purification and regained by the addition of crude extract of S. lividans in the reaction mixture. This effect was confirmed to due to some low-molecular weight component(s) in the crude extract, because the addition of dialyzed crude extract could not recover the enzymatic activity.
Keyword
refolding conditionsaklavinone 11-hydroxylase
ISSN
0257-2389
Publisher
South Korea
Type
Article
Appears in Collections:
Ochang Branch Institute > Anticancer Agent Research Center > 1. Journal Articles
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