Thomas Reeba, Fukamizo Tamo, Suginta Wipa
School of Biomolecular Science and Engineering (BSE), Vidyasirimedhi Institute of Science and Technology (VISTEC), Payupnai, Wangchan District, Rayong, 21210, Thailand.
Bioresour Bioprocess. 2022 Aug 20;9(1):86. doi: 10.1186/s40643-022-00574-8.
Marine Vibrio species are natural degraders of chitin and usually secrete high levels of chitinolytic enzymes to digest recalcitrant chitin to chitooligosaccharides. This study used an endochitinase (VhChiA) from Vibrio campbellii to produce high-quality chitobiose from crustacean chitins. The enzyme was shown to be fully active and stable over 24 h when BSA was used as an additive. When different chitin sources were tested, VhChiA preferentially digested shrimp and squid (α) chitins compared to crab (β) chitin and did not utilize non-chitin substrates. The overall yields of chitobiose obtained from small-scale production using a single-step reaction was 96% from shrimp, and 91% from squid pen and crab-shell chitins. Larger-scale production yielded 200 mg of chitobiose, with > 99% purity after a desalting and purification step using preparative HPLC. In conclusion, we report the employment of an in-house produced chitinase as an effective biocatalyst to rapidly convert chitin food wastes to chitobiose, in a quantity and quality suitable for use in research and commercial purposes. Chitobiose production by this economical and eco-friendly approach can be easily scaled up to obtain multi-gram quantities of chitobiose for chemo-enzymic synthesis of rare chitooligosaccharide derivatives and long chain chitooligosaccharides, as well as preparation of sugar-based functionalized nanomaterials.
海洋弧菌是几丁质的天然降解菌,通常会分泌高水平的几丁质分解酶,将难降解的几丁质消化为壳寡糖。本研究使用了坎氏弧菌的一种内切几丁质酶(VhChiA),从甲壳类几丁质中生产高质量的壳二糖。当使用牛血清白蛋白作为添加剂时,该酶在24小时内表现出完全活性且稳定。在测试不同的几丁质来源时,与蟹(β)几丁质相比,VhChiA优先消化虾和鱿鱼(α)几丁质,且不利用非几丁质底物。通过单步反应进行小规模生产时,从虾中获得的壳二糖总产率为96%,从鱿鱼笔和蟹壳几丁质中获得的为91%。大规模生产得到了200毫克壳二糖,在使用制备型高效液相色谱进行脱盐和纯化步骤后,纯度>99%。总之,我们报道了使用自制的几丁质酶作为一种有效的生物催化剂,将几丁质食物废料快速转化为壳二糖,其数量和质量适合用于研究和商业目的。通过这种经济环保的方法生产壳二糖可以很容易地扩大规模,以获得数克量的壳二糖,用于稀有壳寡糖衍生物和长链壳寡糖的化学酶合成,以及制备糖基功能化纳米材料。