期刊
METABOLIC ENGINEERING
卷 9, 期 3, 页码 293-303出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2007.02.001
关键词
methylmalonyl-CoA mutase (MCM); metabolic engineering; Saccharopolyspora erythraea; erythromycin; precursor; strain improvement
资金
- NIGMS NIH HHS [R43 GM063278, R43 GM058943-01A1, R44GM063278, R44 GM063278, R44GM58943, R43 GM063278-01] Funding Source: Medline
Engineering of the methylmalonyl-CoA (mmCoA) metabolite node of the Saccharopolyspora erythraea wild-type strain through duplication of the mmCoA mutase (MCM) operon led to a 50% increase in erythromycin production in a high-performance oil-based fermentation medium. The MCM operon was carried on a 6.8 kb DNA fragment in a plasmid which was inserted by homologous recombination into the S. erythraea chromosome. The fragment contained one uncharacterized gene, ORFI; three MCM related genes, mutA, mutB, meaB: and one gntR-family regulatory gene, mutR. Additional strains were constructed containing partial duplications of the MCM operon, as well as a knockout of ORFI. None of these strains showed any significant alteration in their erythromycin production profile. The combined results showed that increased erythromycin production only occurred in a strain containing a duplication of the entire MCM operon including mutR and a predicted stem-loop structure overlapping the 3' terminus of the mutR coding sequence. (c) 2007 Elsevier Inc. All rights reserved.
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