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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (8): 73-78    DOI:
    
Construction of Alcohol-producing Escherichia coli strains and Improvement of Biological Activity Detection Method for the Engineered Strains
JIANG Jin-ju1,3, ZHANG Hong-yu1,3, REN Bao-yong2,3, SHI Fei1,3, CUI Yu-lin2,3, QIN Song
1. Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences,Yantai 264003,China;
2. Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China;
3. Graduate University of Chinese Academy of Sciences,Beijing 100049,China
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Abstract  

The genes of adhB and pdc,coding ADHII and PDC respectively,are key genes of alcohol production pathway in Zymomonas mobilis.The following vectors,pUC-adhB,pUC-pdc and pUC-adhB-pdc,were created by inserting adhB and pdc into the plasmid pUC18. Both adhB and pdc have a ribosome binding site of rbcLS of Synechococcus sp.PCC7942. The results of biological activity detections showed that ribosome binding sites of rbcLS of PCC7942 can effectively induce the expression of adhB and pdc in E.coli. Jar fermentation showed that the recombined E.coli's ability of ethanol production was obviously improved compared with the initial strain. Aldehyde indicator plate was displaced by a new method for Biological activity detection because of its defects.The new method was as follows. Induced expression of the genes by IPTG,add substrates for the enzymes ADHII and PDC,cultivate for 0.5h to 1h at 37℃ and then add Schiff reagent.The results show that the improved method is more simple and reliable than Aldehyde indicator plate.

Key words： Fuel alcohol      Pyruvate decarboxylase(PDC)      Alcohol Dehydrogenase(ADH)      Aldehyde indicator plate      Zymomonas mobilis      Synechococcus sp. PCC7942     
Received: 11 May 2011      Published: 25 August 2011
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JIANG Jin-ju, ZHANG Hong-yu, REN Bao-yong, SHI Fei, CUI Yu-lin, QIN Song. Construction of Alcohol-producing Escherichia coli strains and Improvement of Biological Activity Detection Method for the Engineered Strains. China Biotechnology, 2011, 31(8): 73-78.

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https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I8/73


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