High-level Expression of an Endoxylanase Gene From Bacillus Sp. In Bacillus Subtilis DB104 for the Production of Xylobiose From Xylan

Appl Microbiol Biotechnol. 1998 Jul;50(1):113-8. doi: 10.1007/s002530051264.

Abstract

To produce xylobiose from xylan, high-level expression of an endoxylanase gene from Bacillus sp. was carried out in Bacillus sabtilis DB104. A 1.62-kb SmaI DNA fragment, coding for an endoxylanase of Bacillus sp., was ligated into the Escherichia coli/B. subtilis shuttle vector pJH27 delta 88, producing pJHKJ4, which was subsequently transformed into B. subtilis DB104. A maximum endoxylanase activity of 105 U/ml was obtained from the supernatant of B. subtilis DB104 harboring pJHKJ4. The endoxylanase was purified to homogeneity by ion-exchange chromatography and the production profile of xylooligosaccharides from xylan by the endoxylanase was examined by HPLC with a carbohydrate analysis column. Xylobiose was the major product from xylan at 40 degrees C and its proportion in the xylan hydrolyzates increased with the reaction time; at 12 h, over 60% of the reaction products was xylobiose. These results suggest that xylobiose, which has a stimulatory effect on the selective growth of the intestinal bacterium Bifidobacterium, can be mass-produced effectively by the endoxylanase of Bacillus sp. cloned in B. subtilis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / enzymology
  • Bacillus subtilis / genetics*
  • Bacillus subtilis / metabolism
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Congo Red / chemistry
  • DNA, Bacterial / chemistry
  • Disaccharides / biosynthesis*
  • Electrophoresis, Agar Gel
  • Electrophoresis, Polyacrylamide Gel
  • Endo-1,4-beta Xylanases
  • Escherichia coli / chemistry
  • Gene Expression Regulation, Bacterial*
  • Gene Expression Regulation, Enzymologic
  • Time Factors
  • Xylans / chemistry*
  • Xylosidases / chemistry
  • Xylosidases / genetics*

Substances

  • DNA, Bacterial
  • Disaccharides
  • Xylans
  • Congo Red
  • Xylosidases
  • Endo-1,4-beta Xylanases
  • xylobiose