期刊
JOURNAL OF BIOTECHNOLOGY
卷 110, 期 1, 页码 95-103出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2004.02.001
关键词
Escherichia coli; 6-deoxyerythronolide B; polyketide production; fed-batch bioprocess
A robust high cell-density fed-batch bioprocess was developed for the heterologous production of 6-deoxyerythronolide B (6-dEB), the macrocyclic core of the antibiotic erythromycin, with a recombinant Escherichia coli strain expressing the 6-deoxyerythronolide B synthase (DEBS) from Saccharopolyspora erythraea. Initial evaluation of the E. coli strain in a 5-l bioreactor with the addition of exogenous propionate for polyketide biosynthesis resulted in a maximum cell density of 30 gl(-1) (OD600 similar to 60) and the production of 700 mg l(-1) of 6-dEB. Retention of the two plasmids harboring the heterologous genes was maintained between 90 and 100% even in the absence of antibiotic selection. However, the accumulation of excess ammonia. in the culture medium was found to significantly decrease the productivity of the cells. Through optimization of the medium composition and fermentation conditions, the maximum cell density was increased by two-fold, and a final titer of 1.1 gl(-1) of 6-dEB was achieved. This represents an 11-fold improvement compared to the highest reported titer of 100 mgl(-1) with E. coli as the production host. (C) 2004 Elsevier B.V All rights reserved.
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