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 journalArticle

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 languageEnglish (US)
Pages (from-to)1484-1490
Number of pages7
JournalBiochemistry (Moscow)
Volume75
Issue number12
DOIs
StatePublished - Dec 2010
Externally publishedYes

Keywords

  • Bacillus psychrodurans
  • DNA methyltransferase
  • enzyme kinetics

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