Publication:
Purification, characterization, and overexpression of an endo-1,4-β-mannanase from thermotolerant Bacillus sp. SWU60

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Issued Date

2017

Resource Type

Article

File Type

application/pdf

ISSN

9593993

DOI

10.1007/s11274-017-2224-7

Other identifier(s)

2-s2.0-85013287779

Rights Holder(s)

Scopus

Bibliographic Citation

World Journal of Microbiology and Biotechnology. Vol 33, No.3 (2017)

Suggested Citation

Seesom W., Thongket P., Yamamoto T., Takenaka S., Sakamoto T., Sukhumsirichart W. Purification, characterization, and overexpression of an endo-1,4-β-mannanase from thermotolerant Bacillus sp. SWU60. World Journal of Microbiology and Biotechnology. Vol 33, No.3 (2017). doi:10.1007/s11274-017-2224-7 Retrieved from: https://hdl.handle.net/20.500.14740/4234

Title

Purification, characterization, and overexpression of an endo-1,4-β-mannanase from thermotolerant Bacillus sp. SWU60

Author(s)

Seesom W.
 Thongket P.
 Yamamoto T.
 Takenaka S.
 Sakamoto T.
 Sukhumsirichart W.

Abstract

Endo-β-1,4-mannanases are important catalytic agents in several industries. The enzymes randomly cleave the β-1,4-linkage in the mannan backbone and release short β-1,4-mannooligosaccharides and mannose. In the present study, mannanase (ManS2) from thermotolerant Bacillus sp. SWU60 was purified, characterized, and its gene was cloned and overexpressed in Escherichia coli. ManS2 was purified from culture filtrate (300 ml) by using hydrophobic, ion-exchange, and size-exclusive liquid chromatography. The apparent molecular mass was 38 kDa. Optimal pH and temperature for enzyme activity were 6.0 and 60 °C, respectively. The enzyme was stable up to 60 °C for 1 h and at pH 5–9 at 4 °C for 16 h. Its enzyme activity was inhibited by Hg2+. The full-length mans2 gene was 1,008 bp, encoding a protein of 336 amino acids. Amino acid sequence analysis revealed that it belonged to glycoside hydrolase family 26. Konjac glucomannan was a favorable substrate for recombinant ManS2 (rManS2). rManS2 also degraded galactomannan from locust bean gum, indicating its potential for production of glucomanno- and galactomanno-oligosaccharides. Both native and recombinant ManS2 from Bacillus sp. SWU60 can be applied in several industries especially food and feed. © 2017, Springer Science+Business Media Dordrecht.

Subject(s)

Amino acids
 Bacteriology
 Cloning
 Enzymes
 Escherichia coli
 Filtration
 Gene encoding
 Genes
 Hydrophobic chromatography
 Ion exchange
 Liquid chromatography
 Purification
 Bacillus sp
 Galactomannans
 Glucomannan
 Mannan
 Recombinant enzymes
 Enzyme activity
 Beta mannosidase
 Galactomannan
 Mannan
 Bacillus
 Biosynthesis
 Chemistry
 Enzyme activation
 Enzyme specificity
 Enzyme stability
 Enzymology
 Escherichia coli
 Genetics
 Ion exchange chromatography
 Isolation and purification
 Metabolism
 Nucleotide sequence
 Procedures
 Bacillus
 Base Sequence
 Beta-Mannosidase
 Chromatography, Ion Exchange
 Enzyme Activation
 Enzyme Stability
 Escherichia coli
 Mannans
 Substrate Specificity

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URI

https://hdl.handle.net/20.500.14740/4234

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Scopus: Year 1983-2021

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