Recombinant DNA-methyltransferase M1.BspACI from Bacillus psychrodurans AC: Purification and properties

被引：0
|
作者：
M. V. Tarasova
V. V. Kuznetsov
N. A. Netesova
D. A. Gonchar
S. Kh. Degtyarev
机构：
[1] SibEnzyme,
[2] Novosibirsk State University,undefined
[3] State Research Center of Virology and Biotechnology “Vector”,undefined
来源：
Biochemistry (Moscow) | 2010年 / 75卷
关键词：
DNA methyltransferase; enzyme kinetics;
D O I：
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中图分类号：
学科分类号：
摘要：
A restriction-modification system from Bacillus psychrodurans AC (recognition sequence 5′-CCGC-3′) comprises two DNA methyltransferases: M1.BspACI and M2.BspACI. The bspACIM1 gene was cloned in the pJW2 vector and expressed in Escherichia coli cells. High-purity M1.BspACI preparation has been obtained by chromatography on different carriers. M1.BspACI has a temperature optimum of 30°C and demonstrates maximum activity at pH 8.0. M1.BspACI modifies the first cytosine in the recognition sequence 5′-CCGC-3′. The kinetic parameters of M1.BspACI DNA methylation are as follows: Km for phage λ DNA is 0.053 μM and Km for S-adenosyl-L-methionine is 5.1 μM. The catalytic constant (kcat) is 0.095 min−1.
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页码：1484 / 1490
页数：6
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