Please use this identifier to cite or link to this item: https://ir.swu.ac.th/jspui/handle/123456789/13499
ชื่อเรื่อง: Identification and characterization of GH62 bacterial α-L-arabinofuranosidase from thermotolerant Streptomyces sp. SWU10 that preferentially degrades branched L-arabinofuranoses in wheat arabinoxylan
ผู้แต่ง: Phuengmaung P.
Kunishige Y.
Sukhumsirichart W.
Sakamoto T.
Keywords: 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
วันที่เผยแพร่: 2018
บทคัดย่อ: 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.
URI: https://ir.swu.ac.th/jspui/handle/123456789/13499
https://www.scopus.com/inward/record.uri?eid=2-s2.0-85041418557&doi=10.1016%2fj.enzmictec.2018.01.009&partnerID=40&md5=4bf7ae0d448f49e4b810def74032a14f
ISSN: 1410229
Appears in Collections:Scopus 1983-2021

Files in This Item:
There are no files associated with this item.


Items in SWU repository are protected by copyright, with all rights reserved, unless otherwise indicated.