Journal of South China University of Technology(Natural Science Edition) ›› 2012, Vol. 40 ›› Issue (5): 84-89.

• Biological Engineering • Previous Articles     Next Articles

Effective Expression of Lipase in Non-Spore Aspergillus niger via Agrobacterium tumefaciens-Mediated Transformation

Pan Li  Wang Yun-yan  Wang Bin  He Pan  Li Jun-xing   

  1. School of Biological Science and Engineering,South China University of Technology,Guangzhou 510006,Guangdong,China
  • Received:2011-09-26 Revised:2011-11-29 Online:2012-05-25 Published:2012-03-31
  • Contact: 潘力(1967-) ，男，教授，博士生导师，主要从事工业微生物遗传生理及应用研究． E-mail:btlipan@scut.edu.cn
  • About author:潘力(1967-) ，男，教授，博士生导师，主要从事工业微生物遗传生理及应用研究．
  • Supported by:

    国家自然科学基金资助项目( 30970045)

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Abstract:

Aspergillus niger ( A. niger) owns effective capabilities of protein expression and secretion. In order to implement the effective recombinant expression of heterologous protein in non-spore A. niger,several selection markers were evaluated and hygromycin B resistance gene was selected to establish an Agrobacterium tumefaciens ( A. tumefaciens) -mediated transformation system of A. niger. Then,Rhizomucor miehei lipase ( RML) was transformed into A. niger SH-1 via the A. tumefaciens-mediated transformation and was verified by means of PCR identification,enzyme activity assay,Ni affinity chromatography and SDS-PAGE. Finally,RML transformants effectively expressing RML protein were obtained after the fermentation optimization. The p-nitrophenol colorimetry results show that the transformants possesses a lipase activity of 15 U/mL,which is more than ten times that expressed in yeast.

Key words: non-spore Aspergillus niger, Agrobacterium tumefaciens-mediated transformation, Rhizomucor miehei lipase, gene expression

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