Publication:
Identification and characterization of the first β-1,3-D-xylosidase from a gram-positive bacterium, Streptomyces sp. SWU10

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Issued Date

2018

Resource Type

Article

File Type

application/pdf

ISSN

1410229

DOI

10.1016/j.enzmictec.2017.11.002

Other identifier(s)

2-s2.0-85033579354

Rights Holder(s)

Scopus

Bibliographic Citation

Enzyme and Microbial Technology. Vol 112, (2018), p.72-78

Suggested Citation

Phuengmaung P., Fujiwara D., Sukhumsirichart W., Sakamoto T. Identification and characterization of the first β-1,3-D-xylosidase from a gram-positive bacterium, Streptomyces sp. SWU10. Enzyme and Microbial Technology. Vol 112, (2018), p.72-78. doi:10.1016/j.enzmictec.2017.11.002 Retrieved from: https://hdl.handle.net/20.500.14740/5348

Title

Identification and characterization of the first β-1,3-D-xylosidase from a gram-positive bacterium, Streptomyces sp. SWU10

Author(s)

Phuengmaung P.
 Fujiwara D.
 Sukhumsirichart W.
 Sakamoto T.

Abstract

In previous reports, we characterized four endo-xylanases produced by Streptomyces sp. strain SWU10 that degrade xylans to several xylooligosaccharides. To obtain a set of enzymes to achieve complete xylan degradation, a β-D-xylosidase gene was cloned and expressed in Escherichia coli, and the recombinant protein, named rSWU43A, was characterized. SWU43A is composed of 522 amino acids and does not contain a signal peptide, indicating that the enzyme is an intracellular protein. SWU43A was revealed to contain a Glyco_hydro_43 domain and possess the three conserved amino acid residues of the glycoside hydrolase family 43 proteins. The molecular mass of rSWU43A purified by Ni-affinity column chromatography was estimated to be 60 kDa. The optimum reaction conditions of rSWU43A were pH 6.5 and 40 °C. The enzyme was stable up to 40 °C over a wide pH range (3.1–8.9). rSWU43A activity was enhanced by Fe2+ and Mn2+ and inhibited by various metals (Ag+, Cd2+, Co2+, Cu2+, Hg2+, Ni2+, and Zn2+), D-xylose, and L-arabinose. rSWU43A showed activity on p-nitrophenyl-β-D-xylopyranoside and p-nitrophenyl-α-L-arabinofuranoside substrates, with specific activities of 0.09 and 0.06 U/mg, respectively, but not on any xylosidic or arabinosidic polymers. rSWU43A efficiently degraded β-1,3-xylooligosaccharides to produce xylose but showed little activity towards β-1,4-xylobiose, with specific activities of 1.33 and 0.003 U/mg, respectively. These results demonstrate that SWU43A is a β-1,3-D-xylosidase (EC 3.2.1.72), which to date has only been described in the marine bacterium Vibrio sp. Therefore, rSWU43A of Streptomyces sp. is the first β-1,3-xylosidase found in gram-positive bacteria. SWU43A could be useful as a specific tool for the structural elucidation and production of xylose from β-1,3-xylan in seaweed cell walls. © 2017 Elsevier Inc.

Subject(s)

Amino acids
 Cloning
 Column chromatography
 Enzyme activity
 Escherichia coli
 Gene encoding
 Hydrolases
 Seaweed
 Sugars
 Xylose
 Amino acid residues
 Glycoside hydrolase family 43
 Gram-positive bacterium
 Intracellular proteins
 Optimum reaction conditions
 Streptomyces
 Structural elucidation
 Xylooligosaccharides
 Recombinant proteins
 4 nitrophenyl alpha arabinofuranoside
 4 nitrophenyl beta arabinofuranoside
 Arabinose
 Bacterial enzyme
 Beta 1,3 dextro xylosidase
 Cadmium
 Cobalt
 Copper
 Glycosidase
 Mercury
 Silver
 Unclassified drug
 Xylose
 Zinc
 Bacterial protein
 Glucuronic acid
 Oligosaccharide
 Recombinant protein
 Xylan
 Xylan endo 1,3 beta xylosidase
 Xylooligosaccharide
 Article
 Column chromatography
 Enzyme activity
 Enzyme analysis
 Enzyme stability
 Matrix assisted laser desorption ionization time of flight mass spectrometry
 Molecular cloning
 Molecular weight
 Nonhuman
 Nucleotide sequence
 PH
 Streptomyces
 Temperature
 Vibrio
 Amino acid sequence
 Bacterial gene
 Biotechnology
 Chemistry
 Enzyme specificity
 Enzymology
 Genetics
 Kinetics
 Metabolism
 Sequence homology
 Streptomyces
 Amino Acid Sequence
 Bacterial Proteins
 Biotechnology
 Cloning, Molecular
 Enzyme Stability
 Genes, Bacterial
 Glucuronates
 Kinetics
 Molecular Weight
 Oligosaccharides
 Recombinant Proteins
 Sequence Homology, Amino Acid
 Streptomyces
 Substrate Specificity
 Xylan Endo-1,3-beta-Xylosidase
 Xylans

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https://hdl.handle.net/20.500.14740/5348

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