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中国生物工程杂志

China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (1): 67-74    DOI: 10.13523/j.cb.20150110
    
The Construction and Application of EchDA Genetic Engineering Bacteria
ZHAO Guo-ling, TAO Xin-yi, WANG Feng-qing, WEI Dong-zhi
State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China
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Abstract  

The EchDA was systematically overexpressed by genetic engineering in the heterologous hosts Streptomyces lividans TK24. Mature peptide coding fragement of EchDA was amplified from Actinoplanesutahensis NRRL 12052. The expression plasmids pNW-S11, pNW-S12 and pNW-S13 are constructed by inserting EchDA,PermE*promoter and PSau3A promoter into vector pNW-S1. As a result, The EchDA was secretion expressed after induction with IPTGand the enzyme activity can reach 125 U/L under the best conditions for industrialized production.With optimization of the reaction conditions with 15 g/Liter FR901379, the molar conversion can achieve 100% within 24 hours.The highest enzyme activity by 300 L fermentation tank can reach 80 U/L, and the molar conversion can achieve 100%with 10 g/L FR901379. EchDA in industry have been relying on fermentationof original producer,Actinoplanesutahensis, which have low yield and activity. The construction of recombinant EchDA producing strain will be must to improve the current production process of echinocandin antifungal drugs.



Key wordsStreptomyces lividans      TK24      Echinocandindeacylase      Genetic engineering Actinoplanesutahensis     
Received: 25 July 2014      Published: 25 January 2015
ZTFLH:  Q786  
Cite this article:

ZHAO Guo-ling, TAO Xin-yi, WANG Feng-qing, WEI Dong-zhi. The Construction and Application of EchDA Genetic Engineering Bacteria. China Biotechnology, 2015, 35(1): 67-74.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150110     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I1/67


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