Abstract
N-acetyl-d-glucosamine (GlcNAc) is an important amino-monosaccharide with great potential for biotechnological applications. It has traditionally been produced by the chemical hydrolysis of chitin, despite certain industrial and environmental drawbacks, including acidic wastes, low yields and high costs. Therefore, enzymatic production has gained attention as a promising environmentally-friendly alternative to the chemical processes. In this study we demonstrate the GlcNAc bioproduction from colloidal α-chitin using an enzyme cocktail containing endochitinases and exochitinases (chitobiosidases and N-acetyl-glucosaminidases). The enzyme cocktail was extracted after fermentation in a bioreactor by Aeromonas caviae CHZ306, a chitinolytic marine bacterium with great potential for chitinase production. Hydrolysis parameters were studied in terms of temperature, pH, enzyme and substrate concentration, and reaction time, achieving over 90% GlcNAc yield within 6 h. The use of colloidal α-chitin as substrate showed a substantial improvement of GlcNAc yields, when compared with β-chitin and α-chitin polymorphs. Such result is directly related to a significant decrease in crystallinity and viscosity from natural α-chitin, providing the chitinase with greater accessibility to the depolymerized chains. This study provides valuable information on the GlcNAc bioproduction from chitin using an enzymatic approach, addressing the key points for its production, including the enzyme cocktail composition and the substrate structures.
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The authors would like to thank the financial support provided from Sao Paulo Research Foundation (FAPESP), Grants 2012/16824–0, 2013/18773–6 and 2016/20970–2
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Electronic supplementary material 1 (PDF 158 kb) HPLC analysis from GlcNAc bioproduction using an enzyme cocktail produced by A. caviae CHZ306. (a) HPLC profile of N-acetylglucosamine and glucosamine standards; (b) HPLC profile of a representative sample of GlcNAc bioproduction; (c) HPLC profile from blank sample
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Cardozo, F.A., Facchinatto, W.M., Colnago, L.A. et al. Bioproduction of N-acetyl-glucosamine from colloidal α-chitin using an enzyme cocktail produced by Aeromonas caviae CHZ306. World J Microbiol Biotechnol 35, 114 (2019). https://doi.org/10.1007/s11274-019-2694-x
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DOI: https://doi.org/10.1007/s11274-019-2694-x