A GH89 human α-N-acetylglucosaminidase (hNAGLU) homologue from gut microbe Bacteroides thetaiotaomicron capable of hydrolyzing heparosan oligosaccharides

AMB Express

Xiaohong Yang¹  Hai Yu¹  Tamashree Ghosh¹  Xiaoxiao Yang¹  Xi Chen¹  John B. McArthur²  Lan Na³  Tsui-Fen Chou⁴  Patricia Dickson⁵ 

[1] Department of Chemistry, University of California, One Shields Avenue, 95616, Davis, CA, USA;Department of Chemistry, University of California, One Shields Avenue, 95616, Davis, CA, USA;Integrated Micro-Chromatography Systems, Inc, 20963, Irmo, SC, USA;Department of Chemistry, University of California, One Shields Avenue, 95616, Davis, CA, USA;Rosalind Franklin Institute and University of Oxford, Harwell Campus, OX11 0FA, Didcot, UK;Division of Biology and Biological Engineering, California Institute of Technology, 91125, Pasadena, CA, USA;Division of Genetics and Genomic Medicine, Department of Pediatrics, Washington University School of Medicine, 63110, St. Louis, MO, USA;

关键词: N;    NAGLU;    Bacterial glycoside hydrolases;    Heparosan oligosaccharides;    Bacteroides thetaiotaomicron;   

DOI  :  10.1186/s13568-021-01253-1

来源: Springer

【 摘 要 】

Carbohydrate-Active enZYme (CAZY) GH89 family enzymes catalyze the cleavage of terminal α-N-acetylglucosamine from glycans and glycoconjugates. Although structurally and mechanistically similar to the human lysosomal α-N-acetylglucosaminidase (hNAGLU) in GH89 which is involved in the degradation of heparan sulfate in the lysosome, the reported bacterial GH89 enzymes characterized so far have no or low activity toward α-N-acetylglucosamine-terminated heparosan oligosaccharides, the preferred substrates of hNAGLU. We cloned and expressed several soluble and active recombinant bacterial GH89 enzymes in Escherichia coli. Among these enzymes, a truncated recombinant α-N-acetylglucosaminidase from gut symbiotic bacterium Bacteroides thetaiotaomicron ∆22Bt3590 was found to catalyze the cleavage of the terminal α1–4-linked N-acetylglucosamine (GlcNAc) from a heparosan disaccharide with high efficiency. Heparosan oligosaccharides with lengths up to decasaccharide were also suitable substrates. This bacterial α-N-acetylglucosaminidase could be a useful catalyst for heparan sulfate analysis.

【 授权许可】

CC BY