Constitutive Overexpression of the Endoxylanase Gene in Bacillus subtilis

  • Kim, Jong-Hyun (Department of Microbiology, Dong-Eui University) ;
  • Kim, Jung-Hoe (Department of Biologicla Sciences, Korea Advanced Institute of Science and Technology) ;
  • Kim, Sun-Chang (Department of Biologicla Sciences, Korea Advanced Institute of Science and Technology) ;
  • Nam, Soo-Wan (Department of Microbiology, Dong-Eui University)
  • Published : 2000.08.01

Abstract

A strong constitutive $P_{JH}$ promoter from Bacillus was applied to overexpress the endoxylanase gene in B. subtilis. The expression plasmid, pHJKJ4, was designed to contain the $P_{JH}$ promoter and endoxylanase promoter ($P_B$), and introduced into B. subtilis DB104. Through batch fermentation of the trasformant cell on a maltose medium, endoxylanase was produced in a growth-associated manner as the predominant protein. The total activity reached about 600 unit/ml at the end of the cultivation, which corresponded to 698 mg endoxylanase protein/l with a specific activity of 860 unit/mg 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.

Keywords

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