Publication:
Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris

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Issued Date

2011

Resource Type

Article

File Type

application/pdf

ISSN

13891723

DOI

10.1016/j.jbiosc.2010.12.024

Other identifier(s)

2-s2.0-79954829587

Rights Holder(s)

Scopus

Bibliographic Citation

Journal of Bioscience and Bioengineering. Vol 111, No.5 (2011), p.528-536

Suggested Citation

Sriyapai T., Somyoonsap P., Matsui K., Kawai F., Chansiri K. Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris. Journal of Bioscience and Bioengineering. Vol 111, No.5 (2011), p.528-536. doi:10.1016/j.jbiosc.2010.12.024 Retrieved from: https://hdl.handle.net/20.500.14740/7334

Title

Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris

Author(s)

Sriyapai T.
 Somyoonsap P.
 Matsui K.
 Kawai F.
 Chansiri K.

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.

Subject(s)

Actinomadura sp. S14
 Family 11
 Hemicellulases
 Pichia Pastoris
 Thermostable xylanase
 Amino acids
 Cloning
 Composting
 Escherichia coli
 Hydrolases
 Purification
 Sugars
 Gene encoding
 Xylan endo 1,3 beta xylosidase
 Actinomadura
 Amino acid sequence
 Article
 Enzyme activity
 Escherichia coli
 Molecular cloning
 Nonhuman
 Nucleotide sequence
 Open reading frame
 Pichia pastoris
 Protein expression
 Protein folding
 Signal transduction
 Thermostability
 Actinomycetales
 Amino Acid Sequence
 Base Sequence
 Cloning, Molecular
 Endo-1,4-beta Xylanases
 Escherichia coli
 Fungal Proteins
 Glycoside Hydrolases
 Hydrogen-Ion Concentration
 Molecular Sequence Data
 Pichia
 Recombinant Proteins
 Substrate Specificity
 Temperature
 Thailand
 Trisaccharides
 Xylans
 Actinomadura sp.
 Escherichia coli
 Pichia pastoris
 Triticum aestivum subsp. spelta

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URI

https://hdl.handle.net/20.500.14740/7334

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Scopus: Year 1983-2021

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