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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (11): 34-38    DOI:
    
Expression of cbhB Gene Driven by Promoter glaA in Aspergillus niger
QIN Hui-bin, YANG Hong-jiang, HUANG Wen, LI Ling-yan
Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457,China
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Abstract  

Aspergillus niger produces three classes of cellulases for cellulose degradation and among these enzymes the expression level of cellobiohydrolases is very low, affecting the cellulose digestion efficiency by the cellulase system of Aspergillus niger.In order to construct an engineered strain with high cellulase enzyme activity, a hybrid gene containing strong promoter fragment of glucoamylase gene glaA and coding region of cellobiohydrolase gene cbhB was constructed via in vitro synthesis. The hybrid gene was subsequently cloned into the binary vector pCAMBIA1301 and the recombinant plasmid was introduced into conidiospores of Aspergillus niger by Agrobacterium tumefaciens-mediated genetic transformation. The transformants were hygromycin resistant with the hybrid gene inserted into chromosome DNA of Aspergillus niger via T-DNA illegitimate recombination. In total, 48 transformants were seleted and their cellulase activities were tested. The results showed that transformant A3-9 had the highest CMC activity which was 1.31-fold as high as the wild type strain of Aspergillus niger, while both transformants B1-7 and A3-6 had the highest FPA activity which was 2.51-fold as high as wild type strain of Aspergillus niger. Furthermore, induction conditions for the hybrid gene expression were also analyzed.



Key wordsAspergillus niger      Cellobiohydrolase gene cbhB      Glucoamylase gene glaA      Hybrid gene     
Received: 26 August 2010      Published: 19 November 2010
ZTFLH:  Q786  
Cite this article:

QIN Hui-bin, YANG Hong-jiang, HUANG Wen, LI Ling-yan. Expression of cbhB Gene Driven by Promoter glaA in Aspergillus niger. China Biotechnology, 2010, 30(11): 34-38.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I11/34

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