期刊
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
卷 90, 期 3, 页码 1171-1177出版社
SPRINGER
DOI: 10.1007/s00253-011-3186-6
关键词
Whole-cell biocatalyst; Aspergillus oryzae; Biodiesel fuel; Transesterification; Fusarium heterosporum lipase
资金
- Ministry of Economy, Trade and Industry (METI)
- MEXT, Japan
In the present study, a system with high lipase expression in Aspergillus oryzae was developed using an improved enolase promoter (P-enoA124) and the 5' , methanolysiuntranslated region of a heat-shock protein (Hsp-UTR). P-enoA142 enhanced the transcriptional level of a heterologous lipase gene and Hsp-UTR improved its translational efficiency. Fusarium heterosporum lipase (FHL) was inserted into a pSENSU-FHL expression vector harboring P-enoA142 and Hsp-UTR and was transformed into an A. oryzae NS4 strain. Transformants possessing pSENSU-FHL in single (pSENSU-FHL#1) and double copies (pSENSU-FHL#2) were selected to evaluate the lipase activity of the whole-cell biocatalyst. The two strains, pSENSU-FHL#1 and #2, showed excellent lipase activity in hydrolysis compared with the strain transformed with conventional expression vector pNAN8142-FHL. Furthermore, by using pSENSU-FHL#2s could proceed much more effectively without deactivation, which allowed a swift addition of methanol to the reaction mixture, thereby reducing reaction time.
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