High-level expression of an endoxylanase gene from Bacillus sp. in Bacillus subtilis DB104 for the production of xylobiose from xylan.
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.
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Taejon, Korea.
K J
KJ
Jeong
I Y
IY
Park
M S
MS
Kim
S C
SC
Kim
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003224:Congo Red / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry; D004187:Disaccharides / Q000096:biosynthesis*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases; D004926:Escherichia coli / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D013997:Time Factors; D014990:Xylans / Q000737:chemistry*; D014995:Xylosidases / Q000737:chemistry / Q000235:genetics*
D004269:DNA, Bacterial; D004187:Disaccharides; D014990:Xylans; D003224:Congo Red; D014995:Xylosidases; D043364:Endo-1,4-beta Xylanases; C004173:xylobiose