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Combining chitinase C and N-acetylhexosaminidase from Streptomyces coelicolor A3(2) provides an efficient way to synthesize N-acetylglucosamine from crystalline chitin

Nguyen-Thi, Nhung; Doucet, Nicolas (2016). Combining chitinase C and N-acetylhexosaminidase from Streptomyces coelicolor A3(2) provides an efficient way to synthesize N-acetylglucosamine from crystalline chitin Journal of Biotechnology , vol. 220 . p. 25-32. DOI: 10.1016/j.jbiotec.2015.12.038.

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Résumé

The enzymatic bioconversion of chitin is of considerable interest for the natural production of bioactive compounds such as chitooligosaccharides and N-acetyl- d-glucosamine (GlcNAc). Key enzymes are involved in the natural processing of chitin, hydrolyzing this abundant biopolymer to yield chitooligosaccharides with substantial value to the medicinal and biotechnological fields. In this study, chitinase C (ScChiC) from the soil bacterium and chitin decomposer Streptomyces coelicolor A3(2) was expressed, purified and characterized. We also optimized a Streptomyces lividans system generating ScChiC expression yields nearly 500-fold higher than the previously reported heterologous expression in Escherichia coli. The purified enzyme was found to be stable below 55. °C for a broad range of pH values (pH 3.5-9) and exhibited high activity against chitin and chitooligosaccharides to form chitobiose (C2) as main product. Crab shell chitin hydrolysis profiles also revealed that ScChiC catalyzes the bioconversion of chitopolysaccharides through an endo-nonprocessive mode of action. When combining ScChiC with an N-acetylhexosaminidase from S. coelicolor A3(2) (ScHEX) in an assay using crude extracts and crystalline chitin as substrate, GlcNAc was generated as final product with a yield over 90% after 8 h incubation. This chitin hydrolysis yield represents one of the most efficient enzyme bioconversion of chitopolysaccharides to GlcNAc characterized to date, making the S. coelicolor ScChiC- ScHEX pair a potentially suitable contender for the viable industrial production of this important bioactive compound.

Type de document: Article
Mots-clés libres: Bioconversion; Biotechnology; Carbohydrates; Enzyme; Glucosamine; Bacteria; Crystalline materials; Enzymes; Escherichia coli; Hydrolysis; Purification; Chitooligosaccharides; Enzymatic bioconversions; Heterologous expression; Industrial production; N-acetyl-D-glucosamine; N-acetylhexosaminidase; Streptomyces coelicolor A3(2); Streptomyces lividans; Chitin; beta n acetylhexosaminidase; chitinase; hydrolase; n acetylglucosamine; Article; biotransformation; enzyme activity; enzyme stability; pH; polymerase chain reaction; priority journal; protein expression; Streptomyces; coelicolor; synthesis
Centre: Centre INRS-Institut Armand Frappier
Date de dépôt: 20 févr. 2019 16:24
Dernière modification: 20 févr. 2019 16:24
URI: http://espace.inrs.ca/id/eprint/4609

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