title: Purification and characterization of xylanase A from Clostridium stercorarium F-9 and a recombinant Escherichia coli.
abstract: Xylanase A encoded by the Clostridium stercorarium F-9 xynA gene was purified to homogeneity from a recombinant clone of Escherichia coli. The N-terminal amino acid sequence and molecular weight (54,000) estimated by SDS-PAGE of the purified enzyme were consistent with those deduced from the nucleotide sequence [Biosci. Biotech. Biochem., 57, 273-277 (1993)]. A xylanase was also purified to homogeneity from a culture supernatant of C. stercorarium F-9. Its N-terminal amino acid sequence, molecular weight, and enzymatic properties were quite in agreement with those of the recombinant enzyme, indicating that the xynA gene was predominantly expressed as a xylanase gene in C. stercorarium F-9. The purified enzyme hydrolyzed xylotriose to yield xylobiose and xylose while it was less active toward xylobiose. It was optimally active at 75 degrees C and pH 7.0 Km and Vmax were estimated to be 1.9 mg/ml and 2.8 mumol of xylose equivalent/min/micrograms for oat spelt xylan, respectively.
authors.affiliation: Department of Bioscience, Faculty of Bioresources, Mie University, Tsu, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sakka
authors.forename: Y
authors.initials: Y
authors.lastname: Kojima
authors.forename: T
authors.initials: T
authors.lastname: Kondo
authors.forename: S
authors.initials: S
authors.lastname: Karita
authors.forename: K
authors.initials: K
authors.lastname: Shimada
authors.forename: K
authors.initials: K
authors.lastname: Ohmiya
mesh_terms: D000595:Amino Acid Sequence; D050260:Carbohydrate Metabolism; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D003013:Clostridium / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping
chemical_list: D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; D043364:Endo-1,4-beta Xylanases