不动杆菌属菌株NCIMB 9871中环己醇分解代谢途径中一种转录激活因子（ChnR）和一种6-氧代己酸脱氢酶（ChnE）的鉴定及其编码基因的定位。
Identification of a transcriptional activator (ChnR) and a 6-oxohexanoate dehydrogenase (ChnE) in the cyclohexanol catabolic pathway in Acinetobacter sp. Strain NCIMB 9871 and localization of the genes that encode them.
作者信息
Iwaki H, Hasegawa Y, Teraoka M, Tokuyama T, Bergeron H, Lau P C
机构信息
Department of Biotechnology, Faculty of Engineering & High Technology Research Center, Kansai University, Suita, Osaka 564-8680, Japan.
出版信息
Appl Environ Microbiol. 1999 Nov;65(11):5158-62. doi: 10.1128/AEM.65.11.5158-5162.1999.
Abstract
We identified chnR, a gene encoding an AraC-XylS type of transcriptional activator that regulates the expression of chnB, the structural gene for cyclohexanone monooxygenase (CHMO) in Acinetobacter sp. strain NCIMB 9871. The gene sequence of chnE, which encodes an NADP(+)-linked 6-oxohexanoate dehydrogenase, the enzyme catalyzing the fifth step of cyclohexanol degradation, was also determined. The gene arrangement is chnB-chnE-chnR. The predicted molecular masses of the three polypeptides were verified by radiolabeling by using the T7 expression system. Inducible expression of cloned chnB in Escherichia coli depended upon the presence of chnR. A transcriptional chnB::lacZ fusion experiment revealed that cyclohexanone induces chnB expression in E. coli, in which a 22-fold increase in activity was observed.
摘要
我们鉴定出了chnR基因，它编码一种AraC-XylS型转录激活因子，该因子可调控不动杆菌属菌株NCIMB 9871中环己酮单加氧酶（CHMO）的结构基因chnB的表达。我们还确定了chnE的基因序列，chnE编码一种与NADP(+)相连的6-氧代己酸脱氢酶，该酶催化环己醇降解的第五步反应。基因排列顺序为chnB-chnE-chnR。通过使用T7表达系统进行放射性标记，验证了这三种多肽的预测分子量。克隆的chnB在大肠杆菌中的诱导表达依赖于chnR的存在。一项转录chnB::lacZ融合实验表明，环己酮可诱导大肠杆菌中chnB的表达，其活性增加了22倍。
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