Department of Microbiology, Biomedical Sciences Institute, University of Sao Paulo, 1374 Professor Lineu Prestes Ave., São Paulo, SP, 05508-000, Brazil.
Sao Carlos Institute of Chemistry, University of Sao Paulo, Trabalhador Sao-Carlense Ave, 400, Sao Carlos, SP, 13560-970, Brazil.
World J Microbiol Biotechnol. 2019 Jul 22;35(8):114. doi: 10.1007/s11274-019-2694-x.
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.
N-乙酰-D-葡萄糖胺(GlcNAc)是一种重要的氨基单糖,具有巨大的生物技术应用潜力。尽管存在某些工业和环境方面的缺点,包括酸性废物、低产率和高成本,但它传统上是通过甲壳素的化学水解来生产的。因此,酶法生产作为一种有前途的替代化学工艺的环保方法引起了人们的关注。在这项研究中,我们使用含有内切几丁质酶和外切几丁质酶(壳二糖酶和 N-乙酰氨基葡萄糖苷酶)的酶混合物,从胶体 α-几丁质中展示了 GlcNAc 的生物生产。该酶混合物是在生物反应器中由具有巨大几丁质酶生产潜力的海洋细菌气单胞菌 CHZ306 发酵后提取的。水解参数在温度、pH 值、酶和底物浓度以及反应时间方面进行了研究,在 6 小时内获得了超过 90%的 GlcNAc 产率。与β-几丁质和α-几丁质多晶型物相比,胶体 α-几丁质作为底物的使用显著提高了 GlcNAc 的产率。这种结果与天然α-几丁质的结晶度和粘度显著降低直接相关,为几丁质酶提供了更大的可达性,以分解聚合链。这项研究提供了关于使用酶法从几丁质生产 GlcNAc 的宝贵信息,解决了生产的关键点,包括酶混合物的组成和底物结构。