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標題: Improved subtilisin YaB production in Bacillus subtilis using engineered synthetic expression control sequences
作者: Wang, J.P.
Yeh, C.M.
Tsai, Y.C.
關鍵字: subtilisin YaB;synthetic expression control sequence;Bacillus subtilis;alkalophilic bacillus;rna-polymerase;alkaline elastase;translational;efficiency;bacterial promoters;gene;binding;construction;recognition;elements
Project: Journal of Agricultural and Food Chemistry
期刊/報告no:: Journal of Agricultural and Food Chemistry, Volume 54, Issue 25, Page(s) 9405-9410.
Alkaline elastase YaB, a favorable meat tenderizer, is an extracellular subtilisin-type protease produced by wild strain alkalophilic Bacillus YaB. The gene ale coding for subtilisin YaB with its own expression control sequence has been cloned and expressed in Bacillus subtilis, but at levels much lower than in the parental strain Bacillus YaB. This study investigates the influence of various expression control sequences including expression control sequences of cdd and veg from B. subtilis, a synthetic expression control sequence ( SECS), and engineered synthetic expression control sequences ( engineered SECSs) on the expression of subtilisin YaB in B. subtilis. The engineered SECSs were generated by using the Polymerase Chain Reaction; their UP element, Shine-Dargarno (SD) sequence, or both were different from those of the native SECS. The expression efficiencies of SECS and engineered SECSs were higher than those of expression control sequences of ale, cdd, and veg. Substitution of the SD sequence of SECS resulted in higher expression of subtilisin YaB than substitution of the UP element, whereas combined substitution of both gave the highest expression. These results demonstrate that engineering of SECSs is an approach for improving subtilisin YaB production in B. subtilis. Moreover, it is suggested that these enginnered SECSs could potentially be used to express homologous and heterologous proteins in B. subtilis at high level.
ISSN: 0021-8561
DOI: 10.1021/jf061982f
Appears in Collections:食品暨應用生物科技學系

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