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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2013, Vol. 33 Issue (8): 67-74    DOI:
    
Engineering E.coli for Isobutanol Production by Two-promoter Vectors
ZHENG Li-juan, CHEN Shao-yun, XU Gang, WU Jian-ping, YANG Li-rong
Zhejiang University, Hangzhou 310027, China
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Abstract  In order to engineer E.coli for efficient isobutanol production, the two-promoter plasmids, pCDFDuet, pRSFDuet and pACYCDuet were utilized to coexpress the isobutanol biosynthetic enzymes. These plasmids are compatible, and each has two T7 promoters. The kivd and alsS genes were coexpressed using the plasmid pCDFDuet. The yqhD and ilvCD genes were coexpressed using the plamid pRSFDuet or pACYCDuet. Thus, two coexpression systems were constructed, resulting in recombinant E. coli named as Eco(CDF+RSF) and Eco(CDF+ACYC), respectively. Eco(CDF+RSF) produced isobutanol 2.7g/L, while Eco(CDF+ACYC) produced more, up to 3.5g/L. The analysis of expression level and activity of each enzyme showed that AHAS and KDCA were expressed more efficiently in Eco(CDF+ACYC), suggesting their important roles in enhanced isobutanol production. In conclusion, the recombinant E. coli Eco(ACYC+CDF) was successfully constructed using two-promoter vectors. It had high isobutanol productivity and was suitable for further engineering for industrial application.

Key words： Isobutanol      E.coli      two-T7 promoter vector      Coexpression     
Received: 11 April 2013      Published: 25 August 2013
ZTFLH:  Q785  
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ZHENG Li-juan
CHEN Shao-yun
XU Gang
WU Jian-ping
YANG Li-rong
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ZHENG Li-juan, CHEN Shao-yun, XU Gang, WU Jian-ping, YANG Li-rong. Engineering E.coli for Isobutanol Production by Two-promoter Vectors. China Biotechnology, 2013, 33(8): 67-74.

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https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2013/V33/I8/67

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