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Title: Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris
Authors: Sriyapai T.
Somyoonsap P.
Matsui K.
Kawai F.
Chansiri K.
Keywords: Actinomadura sp. S14
Family 11
Pichia Pastoris
Thermostable xylanase
Amino acids
Escherichia coli
Gene encoding
xylan endo 1,3 beta xylosidase
amino acid sequence
enzyme activity
Escherichia coli
molecular cloning
nucleotide sequence
open reading frame
Pichia pastoris
protein expression
protein folding
signal transduction
Amino Acid Sequence
Base Sequence
Cloning, Molecular
Endo-1,4-beta Xylanases
Escherichia coli
Fungal Proteins
Glycoside Hydrolases
Hydrogen-Ion Concentration
Molecular Sequence Data
Recombinant Proteins
Substrate Specificity
Actinomadura sp.
Escherichia coli
Pichia pastoris
Triticum aestivum subsp. spelta
Issue Date: 2011
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 Km values toward various xylans, but XynS14 (P. pastoris) showed higher Vmax and Kcat 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. © 2010 The Society for Biotechnology, Japan.
ISSN: 13891723
Appears in Collections:SCOPUS 1983-2021

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