Cloning, expression, and transglycosylation reaction of Paenibacillus sp. strain W-61 xylanase 1.
A xylanase gene, xyn1, which encodes Paenibacillus sp. strain W-61 xylanase 1 (Xyn1), was cloned in Escherichia coli. xyn1 encodes 211 amino acid residues, including 28 amino acid residues of a signal peptide. The deduced amino acid sequence of the mature Xyn1 showed 95.7%, 84.0%, and 83.7% identity to family 11 xylanases of Aeromonas caviae ME-1, Paenibacillus sp., and Bacillus stearothermophilus respectively. The physico-chemical properties of recombinant Xyn1 were very similar to those of intact Xyn1, except for the molecular mass. The pattern of xylooligosaccharides generated by rXyn1 was investigated by fluorophore-assisted carbohydrate electrophoresis (FACE). The degradation rate of xylohexaose by rXyn1 increased markedly as compared with that of xylopentaose. Xylohexaose had a single preferential point of cleavage by rXyn1. On the basis of the pattern of action of xylooligosaccharides, the number of subsites was estimated to be six. The catalytic site was located between the third and the fourth subsites from non-reducing end.
Watanabe
Seiji
S
Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981-8555, Japan.
Viet
Dung Nguyen
DN
Kaneko
Jun
J
Kamio
Yoshiyuki
Y
Yoshida
Shigeki
S
D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004586:Electrophoresis; D015964:Gene Expression Regulation, Bacterial; D006031:Glycosylation; D007700:Kinetics; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein; D014995:Xylosidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D009844:Oligosaccharides; D011994:Recombinant Proteins; D014995:Xylosidases