期刊
BIOCHIMIE
卷 95, 期 7, 页码 1487-1493出版社
ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
DOI: 10.1016/j.biochi.2013.03.014
关键词
Calcium; Divalent ionophore
资金
- National Science Foundation of China
- Ministry of Science and Technology [2012CB721004]
- Ministry of Education of China
- Shanghai Municipal Council of Science and Technology
- Shanghai Leading Academic Discipline Project [B203]
- State Key Laboratory of Bio-organic
- Natural Products Chemistry (CAS)
- National Program of Development of Transgenic New Species of China
Calcimycin is a rare divalent cation specific ionophore antibiotic that has many biochemical and pharmaceutical applications. We have recently cloned and sequenced the Streptomyces chartreusis calcimycin biosynthesis gene cluster as well as identified the genes required for the synthesis of the polyketide backbone of calcimycin. Additional modifying or decorating enzymes are required to convert the polyketide backbone into the biologically active calcimycin. Using targeted mutagenesis of Streptomyces we were able to show that calM from the calcimycin biosynthesis gene cluster is required for calcimycin production. Inactivating calM by PCR targeting, caused high level accumulation of N-demethyl calcimycin. CalM in the presence of S-adenosyl-L-methionine converted N-demethyl calcimycin to calcimycin in vitro. The enzyme was determined to have a kinetic parameter of K-m 276 mu M, k(cat) 1.26 min(-1) and k(cat)/K-m 76.2 M-1 s(-1). These results proved that CalM is a N-methyltransferase that is required for calcimycin biosynthesis, and they set the stage for generating much desired novel calcimycin derivatives by rational genetic and chemical engineering. (C) 2013 Elsevier Masson SAS. All rights reserved.
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