Cited by 35Enzyme Production and CharacterizationCarbohydrate Chemistry and SynthesisGlycosylation and Glycoproteins Research

Cloning, characterization, and production of three α‐l‐fucosidases from Clostridium perfringens ATCC 13124

Fan, Shuquan, Zhang, Huaqin, Chen, Xiaodi, Lu, Lili, Xu, Li, Xiao, Min·2016·Journal of Basic Microbiology

Abstract

α‐l‐Fucosidases are key enzymes for the degradation of intestinal glycans by gut microbes. In this work, three putative α‐l‐fucosidases (Afc1, Afc2, and Afc3) genes from Clostridium perfringens ATCC 13124 were cloned and expressed in Escherichia coli. Afc1 had the α‐l‐fucosidase domain of glycoside hydrolase (GH) 29 family but showed no enzyme activity toward all the substrates examined. The putative acid/base residue of Afc1, Ser205, was replaced by a glutamic acid which is conserved in GH29‐B α‐l‐fucosidases. However, the mutant Afc1‐S205E still failed to show enzyme activity. Afc2 and Afc3 were determined to be 1,3‐1,4‐α‐l‐fucosidase of GH29‐B subfamily and 1,2‐α‐l‐fucosidase of GH95 family, respectively, and both of them could release fucose from porcine gastric mucin (PGM). When C. perfringens ATCC 13124 grew with the presence of PGM, the transcription of afc 1 decreased slightly, while those of afc 2 and afc 3 increased to 2.2‐fold and 1.4‐fold, respectively, and the enzyme activities of Afc2 and Afc3 in the culture increased to 2.2‐fold and 2.6‐fold, respectively. These results suggest that Afc2 and Afc3 are involved in the degradation of intestinal fucosyl glycans by C. perfringens ATCC 13124.

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Crossref · 10.1002/jobm.201500582

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Authors
Fan, S., Zhang, H., Chen, X., Lu, L., Xu, L., & Xiao, M.
Affiliations
Shandong University (CN)
Corresponding author
Xiao, Min
Venue
Journal of Basic Microbiology
Publisher
Wiley
Published
2015-12-11
Volume
56
Issue
4
Pages
347-357
ISSN
  • 0233-111X (ISSN-L)
  • 1521-4028
OpenAlex
W2195422852
Cited by
35
References
45
Citations by year
2025: 4 · 2024: 6 · 2023: 4 · 2022: 5 · 2020: 5 · 2019: 5 · 2018: 1 · 2017: 4 · 2016: 1
FWCI
1.66
Type
journal article
Language
en
Open access
closed
Locations
  • Journal of Basic Microbiology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Major State Basic Research Development Program of China (2012CB822102)
  • National High Technology Research and Development Program of China (2012AA021504)
  • Key grant Project of Chinese Ministry of Education (313033)
  • Science and Technology Development Project of Shandong Province (2014GSF121006)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
Keywords
alpha-L-fucosidase, Clostridium perfringens, porcine gastric mucin, gene cloning, recombinant enzyme expression, gene expression regulation, gut microbiota
MeSH terms
Animals, Cloning, Molecular, Clostridium perfringens, Enzyme Activation, Escherichia coli, Fucose, alpha-L-Fucosidase, Gastric Mucins, Polysaccharides, Substrate Specificity, Swine, Mutagenesis, Site-Directed, Glutamic Acid

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