Purification, characterization and amplification of a 1.8 kbp fragment of xylanase 5 from Aeromonas caviae W-61.
Aeromonas caviae W-61 produces multiple extracellular xylanases, the xylanases 1, 2, 3, 4, and 5. In this study, we purified and characterized the xylanase 5 of A. caviae W-61, and amplified a part of xylanase 5 gene (xyn5). The purified xylanase 5 was found to be a single polypeptide with molecular mass of 140 kDa. It was an endo-beta-1,4-xylanase showing optimum temperature 40 degrees C and optimum pH 6.0. Xylobiose, xylotriose, xylotetrose, xylopentose, xylohexose and a small amount of xylose were detected as the hydrolysis products. The N-terminal amino acid sequence and several internal amino acid sequences of xylanases 5 were determined. From the sequence, a 1.8 kbp fragment was amplified by PCR using forward and reverse primers. DNA sequencing confirmed the presence of nucleotide sequences corresponding to the N-terminal amino acid sequence and the internal amino acid sequences of xylanase 5.
Department of Biochemistry, University of Rajshahi, Bangladesh. nroy_64@yahoo.com
Narayan
N
Roy
Yoshiyuki
Y
Kamio
D057585:Aeromonas caviae / Q000201:enzymology*; D000595:Amino Acid Sequence; D018554:Avena; D001483:Base Sequence; D003001:Cloning, Molecular; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity; D013696:Temperature; D014995:Xylosidases / Q000737:chemistry*
D014995:Xylosidases