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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (03): 46-51    DOI:
    
Deletion Analysis of aveD Gene from a Streptomyces avermitilis Mutant Producing Doramectin
1. Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin 300457, China
2.Academy of State Administration of Grain, Beijing 100037, China
3.Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education,Fujian Agriculture and Forestry University, Fuzhou 350002, China
4.Department of Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100094, China
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Abstract  

The avermectin analog doramectin(CHC-B1), sold commercially as DectomexTM, was co-produced with the undesired analog CHC-B2, CHC-A1 and CHC-A2 by Streptomyces avermitilis bkd76-3 with the supplementation of cyclohexanecarboxylic acid(CHC) during fermentation. Gene deletion vector pXJ04(pKC1139::△aveD1+△aveD2) was used to delete aveD gene in S.avermitilis bkd76-3. The aveD gene was displaced by deletion allele on the plasmid via double crossover. Shaking flask experiments and HPLC analysis showed that the aveD deletion mutant no longer produced the undesired analogue CHC-A1 and CHC-A2, and only made two components which were confirmed as CHC-B1 and CHC-B2 by LC/MS analysis. The yield of CHC-B1 improved 78.19%, and B2 improved 602.3%, the ratio of effective component had a 93.16% increase. The deletion mutant was proved to be genetically stable, and thus might be promising strain in industrial production of doramectin.



Key wordsStreptomyces avermitilis      aveD      Gene deletion      Doramectin     
Received: 26 November 2009      Published: 25 March 2010
Cite this article:

WANG Ji-Ying, GAN Qiu-Feng, ZHANG Xiao-Lin, HONG Xiang, SONG Yuan, LU Fu-Beng. Deletion Analysis of aveD Gene from a Streptomyces avermitilis Mutant Producing Doramectin. China Biotechnology, 2010, 30(03): 46-51.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I03/46

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