Publication:
Identification and characterization of GH62 bacterial α-L-arabinofuranosidase from thermotolerant Streptomyces sp. SWU10 that preferentially degrades branched L-arabinofuranoses in wheat arabinoxylan

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Issued Date

2018

Resource Type

Article

File Type

application/pdf

ISSN

1410229

DOI

10.1016/j.enzmictec.2018.01.009

Other identifier(s)

2-s2.0-85041418557

Rights Holder(s)

Scopus

Bibliographic Citation

Enzyme and Microbial Technology. Vol 112, (2018), p.22-28

Suggested Citation

Phuengmaung P., Kunishige Y., Sukhumsirichart W., Sakamoto T. Identification and characterization of GH62 bacterial α-L-arabinofuranosidase from thermotolerant Streptomyces sp. SWU10 that preferentially degrades branched L-arabinofuranoses in wheat arabinoxylan. Enzyme and Microbial Technology. Vol 112, (2018), p.22-28. doi:10.1016/j.enzmictec.2018.01.009 Retrieved from: https://hdl.handle.net/20.500.14740/5744

Title

Identification and characterization of GH62 bacterial α-L-arabinofuranosidase from thermotolerant Streptomyces sp. SWU10 that preferentially degrades branched L-arabinofuranoses in wheat arabinoxylan

Author(s)

Phuengmaung P.
 Kunishige Y.
 Sukhumsirichart W.
 Sakamoto T.

Abstract

We previously described thermotolerant Streptomyces sp. SWU10, which produced four endo-xylanases and one xylosidase able to digest xylan backbones. To achieve arabinoxylan degradation, the swu62A gene was cloned and overexpressed in Escherichia coli, and the recombinant enzyme, termed SWUAbf62A, was characterized. The 438 amino acids of SWUAbf62A revealed Glyco_hydro_62 and closely related with putative α-L-arabinofuranosidases belonging to glycoside hydrolase family 62. SWUAbf62A was purified in two steps, Ni-affinity and size-exclusion column chromatographies, and its molecular mass without signal peptide was determined to be 49 kDa. SWUAbf62A showed optimum activity at pH 5.0 and 50 °C, and more than 70% of its initial enzymatic activity remained after incubation at pH 4.1–10.5, while SWUAbf62A lost all activity after 1 h at 60 °C. SWUAbf62A activity was stimulated by Ba2+, Ca2+, and Mn2+ and decreased by Ag+, Cu2+, Fe2+, and EDTA. SWUAbf62A had no activity towards p-nitrophenyl-α-L-arabinofuranoside or p-nitrophenyl-β-D-xylopyranoside synthetic substrates. On the other hand, SWUAbf62A had the highest activity against wheat arabinoxylan, with a specific activity of 1.29 U/mg, and was also active against sugar beet arabinan, with a specific activity of 0.14 U/mg; these results indicated that SWUAbf62A is an arabinoxylan arabinofuranohydrolase. Using 1H-NMR analysis, SWUAbf62A was found to release L-arabinofuranoses singly linked to O-3 of wheat arabinoxylan. In addition, SWUAbf62A acted synergistically with endo-xylanase (XynSW3) and α-L-arabinofuranosidase, which releases arabinose linked to O-3 of double-substituted xylose residues on arabinoxylan, to digest the wheat arabinoxylan. SWUAbf62A is an important debranching enzyme for hydrolysis of hemicelluloses to monosaccharides and can be applied in various industrial biotechnologies. © 2018 Elsevier Inc.

Subject(s)

Cloning
 Escherichia coli
 Gene encoding
 Hydrolases
 Sugar beets
 Sugars
 Arabinoxylans
 Enzymatic activities
 Glycoside hydrolases
 Industrial biotechnology
 Recombinant enzymes
 Streptomyces
 Synergistic action
 Synthetic substrates
 Enzyme activity
 Alpha arabinofuranosidase
 Arabinose
 Arabinoxylan
 Bacterial enzyme
 Glycosidase
 Hemicellulose
 Monosaccharide
 Recombinant enzyme
 Xylan endo 1,3 beta xylosidase
 Xylose
 Alpha-N-arabinofuranosidase
 Arabinofuranose
 Arabinoxylan
 Bacterial protein
 Polysaccharide
 Recombinant protein
 Xylan
 Article
 Crystallization
 Enzyme activity
 Enzyme assay
 Escherichia coli
 Heat tolerance
 Molecular cloning
 Molecular weight
 Nonhuman
 PH
 Regioselectivity
 Streptomyces
 Streptomyces coelicolor
 Sugar beet
 Wheat
 Amino acid sequence
 Analogs and derivatives
 Bacterial gene
 Biomass
 Biotechnology
 Chemistry
 Enzyme specificity
 Enzymology
 Genetics
 Heat
 Hydrolysis
 Kinetics
 Metabolism
 Sequence homology
 Streptomyces
 Amino Acid Sequence
 Arabinose
 Bacterial Proteins
 Biomass
 Biotechnology
 Genes, Bacterial
 Glycoside Hydrolases
 Hot Temperature
 Hydrolysis
 Kinetics
 Polysaccharides
 Recombinant Proteins
 Sequence Homology, Amino Acid
 Streptomyces
 Substrate Specificity
 Triticum
 Xylans

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URI

https://hdl.handle.net/20.500.14740/5744

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Scopus: Year 1983-2021

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