食品科学 ›› 2022, Vol. 43 ›› Issue (6): 74-80.doi: 10.7506/spkx1002-6630-20210205-106

• 生物工程 • 上一篇    下一篇

双酶协同降解胶体几丁质及其作用机制

吴昊,刘嘉荔,杨静文,胡雪芹,张洪斌   

  1. (合肥工业大学食品与生物工程学院,安徽合肥 230009)
  • 出版日期:2022-03-25 发布日期:2022-03-28
  • 基金资助:
    国家自然科学基金面上项目(81573399); 2019年度合肥工业大学智能制造技术研究院科技成果转化及产业化重点项目(IMICZ2019004); 安徽省自然科学基金项目(1908085QC130)

Synergistic Enzymatic Degradation of Colloidal Chitin and Its Mechanism

WU Hao, LIU Jiali, YANG Jingwen, HU Xueqin, ZHANG Hongbin   

  1. (School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China)
  • Online:2022-03-25 Published:2022-03-28

摘要: 建立一种几丁质酶(chitinase,ChiA)和β-N-乙酰基己糖胺酶HJ5N协同催化体系,可实现一步反应降解胶体几丁质制备N-乙酰氨基葡萄糖(N-acetylglucosamine,GlcNAc),探讨不同因素对双酶协同催化体系的影响和双酶协同作用机制。结果表明,双酶协同催化体系的最适反应温度为30?℃,最适反应pH值为6,ChiA与HJ5N添加比(加酶量(U)计)为2∶1。在最优反应条件下以90?g/L虾壳几丁质(胶体几丁质30.3?g/L)为底物时,GlcNAc产量为17.55?mg/mL,收率达到58%。ChiA与HJ5N的协同作用机制主要表现在HJ5N可保证中间产物几丁二糖(GlcNAc)2快速被转化为GlcNAc,有效解除(GlcNAc)2对ChiA的抑制作用,从而实现双酶协同高效催化降解胶体几丁质制备GlcNAc。这为酶法降解胶体几丁质制备GlcNAc的工业应用提供了一种有效策略。

关键词: 几丁质酶;β-N-乙酰基己糖胺酶;N-乙酰氨基葡萄糖;双酶协同催化;胶体几丁质

Abstract: In this study, a synergistic catalytic system of chitinase (ChiA) and β-N-acetylhexosaminase HJ5N that can degrade colloidal chitin to prepare N-acetylglucosamine (GlcNAc) in one-step reaction was established. The effects of different factors on the catalytic efficiency and the mechanism of the synergistic action of the enzymes were discussed. The results showed that the optimum reaction conditions were 30 ℃, pH 6 and concentration ratio ChiA to HJ5N of 2:1. Under these conditions, 17.55 mg/mL of GlcNAc was produced with a yield of 58% when 90 g/L of (containing 30.3 g/L of colloidal chitin) was used as the substrate. HJ5N could ensure quick conversion of the intermediate product (GlcNAc)2 into GlcNAc, thus removing the inhibitory effect of (GlcNAc)2 on ChiA and ultimately allowing the efficient degradation of colloidal chitin into GlcNAc. This study provides an effective strategy for the industrial application of enzymatic degradation of colloidal chitin to prepare N-acetylglucosamine.

Key words: chitinase; β-N-acetylhexosaminase; N-acetylglucosamine; synergistic enzymatic catalysis; colloidal chitin

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