Recombinant Expression of Rhizopus chinensis Lipase in Aspergillus niger

doi:10.13560/j.cnki.biotech.bull.1985.2015.04.024

Abstract

Abstract: This article carried out the recombinant expression of Rhizopus chinensis lipase(RCL)gene in Aspergillus niger. Promoter of Pgpd from A. niger and RCL-TtrpC fusing fragment were respectively obtained by PCR and further cloned to plasmid pCHAMBIA2302 to generate the RCL over-expression vector pCHAMBIA2302∷Pgpd-RCL-TtrpC. The resulting vector was introduced into Agrobacterium tumefaciens EHA105, and then transformed into A. niger through the mediation of A. tumefaciens. Seven randomly selected transformants were analyzed by PCR and Southern blot. As a result, all the transformants were found to be positive. Then, the seven transformants were subjected to fermentation and lipase assay, and the results showed that all the transformants secreted recombinant RCL, among which transformant T6 produced the highest lipase specific activity, reaching up to approximately 71 U/mL. The fermentation broth was further analyzed by SDS-PAGE, which revealed the molecular weight of the recombinant RCL was 37 kD.

Key words: recombinant expression, Rhizopus chinensis, lipase, Aspergillus niger, Agrobacterium tumefaciens

Zhang Qian, Wang Jianying, Lin Zhi, Jia Jia, Guo Hongtao. Recombinant Expression of Rhizopus chinensis Lipase in Aspergillus niger[J]. Biotechnology Bulletin, 2015, 31(3): 165-170.

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