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KMID : 0545120000100040551
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Journal of Microbiology and Biotechnology
2000 Volume.10 No. 4 p.551 ~ p.553
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Constitutive Overexpression of the Endoxylanase Gene in Bacillus subtilis
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KIM, JONG-HYUN
KIM, JUNG-HOE/KIM, SUN-CHANG/NAM, SOO-WAN
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Abstract
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A strong constitutive P_JH promoter from Bacillus was applied to overexpress the endoxylanase gene in B. subtilis. The expression plasmid, pJHKJ4, was designed to contain the P_JH promoter and endoxylanase promoter (P_B), and introduced into B. subtilis DB 104. Through batch fermentation of the transformant cell on a maltose medium, endoxylanase was produced in a growth-associated manner as the predominant protein. The total activity reached about 600unit/㎖ at the end of the cultivation, which corresponded to 698㎎ endoxylanase protein/l with a specific activity of 860unit/㎎ protein. It was also found that the segregational plasmid instability was less than 30% and most of the endoxylanase activity was detected in the culture medium. This result suggests that the secretory production of endoxylanase can be significantly enhanced with the use of the P_JH promoter and high-cell density culture techniques, quantitatively as well as qualitatively.
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KEYWORD
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