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.
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ジャーナル |
J Biosci Bioeng
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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.
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巻・号 |
111(5)
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ページ |
528-36
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公開日 |
2011-5-1
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DOI |
10.1016/j.jbiosc.2010.12.024
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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 |