RRC ID 11517
著者 Sriyapai T, Somyoonsap P, Matsui K, Kawai F, Chansiri K.
タイトル Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris.
ジャーナル J Biosci Bioeng
Abstract A thermophilic xylan-degrading Actinomadura sp. S14 was isolated from compost in Thailand. Hemicellulase activities such as endo-1,4-β-xylanase, β-xylosidase and α-arabinofuranosidase were induced with xylan-containing agriculture wastes and oat spelt xylan. The gene encoding xylanase consisting of 687bp was cloned from Actinomadura sp. S14. The deduced amino acid sequence contained a signal peptide of 41 amino acids and a probable mature xylanase of 188 amino acids. An open reading frame (xynS14) corresponding to a mature xylanase was expressed in Escherichia coli and Pichia pastoris. The specific activity of purified XynS14 (P. pastoris) was 2.4-fold higher than XynS14 (E. coli). Both XynS14s showed the same basic properties such as optimal pH and temperature (pH 6.0 and 80°C) and stability in a broad pH range (pH 5.0-11.0) and at high temperatures up to 80°C. Both XynS14s showed approximately the same substrate specificity and K(m) values toward various xylans, but XynS14 (P. pastoris) showed higher V(max) and K(cat) than XynS14 (E. coli). Higher specific activities of XynS14 (P. pastoris) may be due to protein-folding in the host. Purified XynS14 showed more endo-1,4-β-xylanase activity on xylan and xylooligosaccharides than on xylotriose.
巻・号 111(5)
ページ 528-36
公開日 2011-5-1
DOI 10.1016/j.jbiosc.2010.12.024
PII S1389-1723(10)00428-7
PMID 21269876
MeSH Actinomycetales / enzymology* Actinomycetales / genetics* Amino Acid Sequence Base Sequence Cloning, Molecular Endo-1,4-beta Xylanases / genetics Endo-1,4-beta Xylanases / metabolism* Escherichia coli / genetics Escherichia coli / metabolism Fungal Proteins / genetics Fungal Proteins / metabolism* Glycoside Hydrolases / genetics Glycoside Hydrolases / metabolism Hydrogen-Ion Concentration Molecular Sequence Data Pichia / genetics Pichia / metabolism Recombinant Proteins / biosynthesis Recombinant Proteins / genetics Substrate Specificity Temperature Thailand Trisaccharides / metabolism Xylans / metabolism
IF 2.366
引用数 48
WOS 分野 FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
一般微生物 JCM 16836