期刊
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
卷 111, 期 5, 页码 528-536出版社
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2010.12.024
关键词
Actinomadura sp S14; Hemicellulases; Thermostable xylanase; Family 11 glycoside hydrolase; Pichia pastoris-expression
资金
- Thailand Research Fund [PHD/0222/2550]
- Srinakharinwirot University [069/2552]
- Japan Society for the Promotion of Science (JSPS)
- Grants-in-Aid for Scientific Research [19101009] Funding Source: KAKEN
A thermophilic xylan-degrading Actinomadura sp. 514 was isolated from compost in Thailand. Hemicellulase activities such as endo-1,4-beta-xylanase, beta-xylosidase and alpha-arabinofuranosidase were induced with xylan-containing agriculture wastes and oat spelt xylan. The gene encoding xylanase consisting of 687 bp 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 colt and Pichia pastoris. The specific activity of purified XynS14 (P. pastoris) was 2.4-fold higher than XynS14 (E. colt). Both XynS14s showed the same basic properties such as optimal pH and temperature (pH 6.0 and 80 degrees C) and stability in a broad pH range (pH 5.0-11.0) and at high temperatures up to 80 degrees 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. colt). Higher specific activities of XynS14 (P. pastoris) may be due to protein-folding in the host. Purified XynS14 showed more endo-1,4-beta-xylanase activity on xylan and xylooligosaccharides than on xylotriose. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.
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