Iso-migrastatin Titer Improvement in the Engineered Streptomyces lividans SB11002 Strain by Optimization of Fermentation Conditions
2010
Wu, Xueyun, Jhejiang Sci-Tech University, Hangzhou, China | Yang, Dong, Jhejiang University, Hangzhou, China | Zhu, Xiangcheng, Jhejiang University, Hangzhou, China | Feng, Zhiyang, University of Wisconsin-Madison, Madison, WI, USA | Lv, Zhengbin, Jhejiang Sci-Tech University, Hangzhou, China | Zhang, Yaozhou, Jhejiang Sci-Tech University, Hangzhou, China | Shen, Ben, University of Wisconsin-Madison, Madison, WI, USA | Xu, Zhinan, Jhejiang University, Hangzhou, China
The heterologous production of iso-migrastatin (iso-MGS) was successfully demonstrated in an engineered S. lividans SB11002 strain, which was derived from S. lividans K4-114, following introduction of pBS11001, which harbored the entire nags biosynthetic gene cluster. However, under similar fermentation conditions, the iso-MGS titer in the engineered strain was significantly lower than that in the native producer - Streptomyces platensis NRRL 18993. To circumvent the problem of low iso-MGS titers and to expand the utility of this heterologous system for iso-MGS biosynthesis and engineering, systematic optimization of the fermentation medium was carried out. The effects of major components in the cultivation medium, including carbon, organic and inorganic nitrogen sources, were investigated using a single factor optimization method. As a result, sucrose and yeast extract were determined to be the best carbon and organic nitrogen sources, resulting in optimized iso-MGS production. Conversely, all other inorganic nitrogen sources evaluated produced various levels of inhibition of iso-MGS production. The final optimized R2YE production medium produced iso-MGS with a titer of 86.5 mg/L, about 3.6-fold higher than that in the original R2YE medium, and 1.5 fold higher than that found within the native S. platensis NRRL 18993 producer.
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