請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/68279
標題: Secreted Production of Renilla Luciferase in Bacillus subtilis
作者: Chiang, C.J.
Chen, P.T.
Chao, Y.P.
關鍵字: Renilla luciferase
secreted protein production
Bacillus subtilis
recombinant nattokinase
molecular-cloning
messenger-rna
stability
expression
vectors
instability
plasmids
proteins
pub110
期刊/報告no:: Biotechnology Progress, Volume 26, Issue 2, Page(s) 589-594.
摘要: Luciferase (Rluc) from the soft coral Renilla reniformis has been widely used as a bioluminescent reporter, and its secreted production has been solely performed in mammalian cells thus far. To make the production more efficient, a series of approaches was attempted to overproduce Rluc extracellularly in Bacillus subtilis. First, Cys124 in the Rluc gene was substituted with Ala. The mutant gene was subsequently incorporated into a pUB110/R6K-based plasmid, consequently, fusing with the P43 promoter and the sacB signal peptide. With the nitrogen-rich medium, B. subtilis strain bearing the plasmid became able to secret a detectable amount of Rluc. Moreover, the secretion signal for the Rluc gene was replaced by the aprN leader peptide with or without the propeptide. The result led to a more than twofold increase in the secreted Rluc. Finally, by enhancing the transcription of the Rluc gene implementing the P43 and spac tandem promoter, it resulted in the secreted Rluc with a yield of 100 mg/L. Overall, this study illustrates a potential strategy for improving the secretion efficiency of heterologous proteins in B. subtilis. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 26: 589-594, 2010
URI: http://hdl.handle.net/11455/68279
ISSN: 8756-7938
文章連結: http://dx.doi.org/10.1002/btpr.351
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