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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (03): 56-60    DOI:
    
Cloning and Expression of Complete Gene for α-amylase From Enriched Metagenomic DNA
1.College of Life Science and Technology, Guangxi University,Nanning 530005,China
2.National Engineering Research Center for Nongrain Biorefinery, Guangxi Academy of Sciences, Nanning 530007, China
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Abstract  

Inverse-nest PCR was applied to clone the full sequence of some α-amylase from enrichment soil metagenomic DNA. The successfully achieved gene(GU045523)showed high homology (Identity:99%)with the partial coding sequence of acid-stable amylase from Bacillus sp. KR-8104. The putative mature peptide gene was inserted into pSE380, a recombinant plasmid pSE380-gxaa was constructed and transformed into Escherichia coli JM109. The purified amylase GXAA showed an optimal activity at pH 7.0 and 75℃,the Km value is 11.6g/L taking soluble starch as substrate. Mutants E27G,A450T and E27G-A450T were constructed, the property showed no remarkable difference from the wild type.



Key wordsMetagenome      Inverse-nest PCR      Flanking unknown sequence      α-Amylase     
Received: 23 December 2009      Published: 25 March 2010
Cite this article:

YANG Jian, CENG Li-Juan, LIAO Sai-Meng, WANG Jing-Yan, DU Li-Qin, HUI Yu-Ta, HUANG Ri-Bei. Cloning and Expression of Complete Gene for α-amylase From Enriched Metagenomic DNA. China Biotechnology, 2010, 30(03): 56-60.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I03/56

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