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

M. V. Tarasova; V. V. Kuznetsov; N. A. Netesova; D. A. Gonchar; S. Kh Degtyarev

doi:10.1134/S0006297910120096

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

M. V. Tarasova, V. V. Kuznetsov, N. A. Netesova, D. A. Gonchar, S. Kh Degtyarev

Research output: Contribution to journal › Article › peer-review

Abstract

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: K _m for phage λ DNA is 0.053 μM and K _m for S-adenosyl-L-methionine is 5.1 μM. The catalytic constant (k _cat) is 0.095 min^-1.

Original language	English (US)
Pages (from-to)	1484-1490
Number of pages	7
Journal	Biochemistry (Moscow)
Volume	75
Issue number	12
DOIs	https://doi.org/10.1134/S0006297910120096
State	Published - Dec 2010
Externally published	Yes

Keywords

Bacillus psychrodurans
DNA methyltransferase
enzyme kinetics

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10.1134/S0006297910120096

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title = "Recombinant DNA-methyltransferase M1.BspACI from Bacillus psychrodurans AC: Purification and properties",

abstract = "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: K m for phage λ DNA is 0.053 μM and K m for S-adenosyl-L-methionine is 5.1 μM. The catalytic constant (k cat) is 0.095 min-1.",

keywords = "Bacillus psychrodurans, DNA methyltransferase, enzyme kinetics",

author = "Tarasova, {M. V.} and Kuznetsov, {V. V.} and Netesova, {N. A.} and Gonchar, {D. A.} and Degtyarev, {S. Kh}",

year = "2010",

month = dec,

doi = "10.1134/S0006297910120096",

language = "English (US)",

volume = "75",

pages = "1484--1490",

journal = "Biochemistry (Moscow)",

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T1 - Recombinant DNA-methyltransferase M1.BspACI from Bacillus psychrodurans AC

T2 - Purification and properties

AU - Tarasova, M. V.

AU - Kuznetsov, V. V.

AU - Netesova, N. A.

AU - Gonchar, D. A.

AU - Degtyarev, S. Kh

PY - 2010/12

Y1 - 2010/12

N2 - 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: K m for phage λ DNA is 0.053 μM and K m for S-adenosyl-L-methionine is 5.1 μM. The catalytic constant (k cat) is 0.095 min-1.

AB - 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: K m for phage λ DNA is 0.053 μM and K m for S-adenosyl-L-methionine is 5.1 μM. The catalytic constant (k cat) is 0.095 min-1.

KW - Bacillus psychrodurans

KW - DNA methyltransferase

KW - enzyme kinetics

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AN - SCOPUS:79251560612

SN - 0006-2979

VL - 75

SP - 1484

EP - 1490

JO - Biochemistry (Moscow)

JF - Biochemistry (Moscow)

IS - 12

ER -

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

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