Publication:
Purification, characterization of GH11 Endo-β-1,4-xylanase from thermotolerant streptomyces sp. SWU10 and overexpression in pichia pastoris KM71H

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Issued Date

2013

Resource Type

Article

File Type

application/pdf

ISSN

10736085

DOI

10.1007/s12033-012-9541-8

Other identifier(s)

2-s2.0-84876155258

Rights Holder(s)

Scopus

Bibliographic Citation

Molecular Biotechnology. Vol 54, No.1 (2013), p.37-46

Suggested Citation

Deesukon W., Nishimura Y., Sakamoto T., Sukhumsirichart W. Purification, characterization of GH11 Endo-β-1,4-xylanase from thermotolerant streptomyces sp. SWU10 and overexpression in pichia pastoris KM71H. Molecular Biotechnology. Vol 54, No.1 (2013), p.37-46. doi:10.1007/s12033-012-9541-8 Retrieved from: https://hdl.handle.net/20.500.14740/6664

Title

Purification, characterization of GH11 Endo-β-1,4-xylanase from thermotolerant streptomyces sp. SWU10 and overexpression in pichia pastoris KM71H

Author(s)

Deesukon W.
 Nishimura Y.
 Sakamoto T.
 Sukhumsirichart W.

Abstract

We have previously described two forms of an endo-β-1,4-xylanase (XynSW2A and XynSW2B) synthesized by thermotolerant Streptomyces sp. SWU10. Here, we describe another xylanolytic enzyme, designated XynSW1. The enzyme was purified to homogeneity from 2 L of culture filtrate. Its apparent molecular mass was 24 kDa. The optimal pH and temperature were pH 5.0 and 40 C, respectively. The enzyme was stable in a wide pH ranges (pH 1-11), more than 80 % of initial activity remained at pH 2-11 after 16 h of incubation at 4 C and stable up to 50 C for 1 h. Xylobiose and xylotriose were the major xylooligosaccharides released from oat spelt xylan by the action of XynSW1, indicating of endo-type xylanase. The complete xynSW1 gene contains 1,011 bp in length and encode a polypeptide of 336 with 41 amino acids of signal peptide. The amino acid sequence analysis revealed that it belongs to glycoside hydrolase family 11 (GH11). The mature xynSW1 gene without signal peptide sequence was overexpressed in Pichia pastoris KM71H. The recombinant XynSW1 protein showed higher molecular mass due to the differences in glycosylation levels at the six N-glycosylation sites in the amino acid sequence and exhibited better physicochemical properties than those of the native enzyme including higher optimal temperature (60 C), and specific activity, but lower optimal pH (4.0). Because of their stability in a wide pH ranges, both of native and recombinant enzymes of XynSW1, may have potential application in several industries including food, textile, biofuel, and also waste treatment. © 2012 Springer Science+Business Media, LLC.

Subject(s)

Amino acid sequence analysis
 GH11
 Glycoside hydrolases
 N-glycosylation sites
 Over-expression
 Physicochemical property
 Pichia Pastoris
 Xylanases
 Amino acids
 Bacteria
 Characterization
 Genes
 Molecular mass
 Optimization
 Polypeptides
 Purification
 Recombinant proteins
 Textile industry
 Waste treatment
 Yeast
 Enzymes
 Biofuel
 Glycosidase
 Recombinant enzyme
 Amino acid sequence
 Article
 Food
 Gene overexpression
 Molecular weight
 Nonhuman
 PH
 Physical chemistry
 Pichia pastoris
 Sequence analysis
 Streptomyces
 Temperature
 Textile
 Waste management
 Amino Acid Sequence
 Cloning, Molecular
 Disaccharides
 Endo-1,4-beta Xylanases
 Gene Expression
 Pichia
 Streptomyces
 Temperature
 Trisaccharides
 Pichia pastoris
 Streptomyces sp.
 Triticum aestivum subsp. spelta

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URI

https://hdl.handle.net/20.500.14740/6664

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Scopus: Year 1983-2021

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