非特异性酶对改性几丁质的降解能力及其产物分析
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(1.河北农业大学食品科技学院 河北保定 071000;2.河北科技师范学院食品科技学院 河北昌黎 066600)

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河北省省级科技计划资助项目(19273201D);河北省现代农业产业技术体系淡水养殖创新团队建设项目(HBCT2018180206)


Degradation Capacity of Non-specific Enzyme for Modified Chitin and Product Analysis
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(1.College of Food Science and Technology, Hebei Agricultural University, Baoding 071000, Hebei;2.College of Food Science and Technology, Hebei Normal University of Science & Technology, Changli 066600, Hebei)

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    摘要:

    由于天然几丁质具有分子质量高,结晶度高,不易酶解的特点,因此其开发和应用受到极大地限制。为了提高几丁质的利用率,筛选具有高降解能力与良好经济效益的商品化非特异性酶,并评价其对经3种改性方法(超微粉碎、高压均质、超微粉碎-高压均质联用)处理的几丁质的酶解效果,确定最佳改性方式,优化酶解工艺,分析酶解产物。结果表明:7种非特异性酶中,木瓜蛋白酶的几丁质降解能力最高,成本最低廉。超微粉碎-高压均质联用法使几丁质具有更高的可降解性。木瓜蛋白酶对几丁质降解的最佳条件:温度50 ℃,pH 7.0,酶质量浓度10 mg/mL,时间14 h。薄层层析结果显示,在酶解初期(1 h)生成(GlcNAc)3-6共4种聚合度的几丁寡糖,初步证明木瓜蛋白酶能够断裂几丁质的β-1,4糖苷键。离子色谱分析结果显示,酶解平衡(14 h)后的酶解产物中只有GlcNAc和(GlcNAc)2,质量浓度分别为22.26 mg/mL和58.75 mg/mL。

    Abstract:

    Natural chitin has the characteristics of high molecular weight, high crystallinity and difficult enzymatic hydrolysis, which greatly limits its development and application. In order to improve the utilization rate of chitin, commercial non-specific enzymes with high degradation ability and good economic benefits were screened in this study, and their enzymatic hydrolysis effects on chitin after three modification methods (ultra-micro grinding, high-pressure homogenization, ultra-micro grinding-high-pressure homogenization) were evaluated. Determine the best modification method, optimize enzymatic hydrolysis process and analyze enzymatic hydrolysis products. The results showed that among the seven non-specific enzymes, papain had the highest chitin degradation ability and the lowest cost. The combined pretreatment of ultra-micro grinding and high-pressure homogenization method made chitin have higher degradability, the optimum conditions for degradation of chitin by papain were temperature 50 ℃, pH 7.0, enzyme mass concentration 10 mg/mL, time 14 h. The results of thin layer chromatography showed that(GlcNAc)3-6 four degrees of polymerization of chitin oligosaccharides were produced at the initial stage of enzymatic hydrolysis(1 h), which preliminarily proved that papain could break the β-1,4 glycosidic bond of chitin; the results of ion chromatography analysis showed that only GlcNAc and (GlcNAc)2 were found in the enzymolysis products after enzymolysis equilibrium (14 h), and the mass concentrations were 22.26 mg/mL and 58.75 mg/mL, respectively.

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刘洋,郭明珠,石文琪,尹朝阳,桑亚新,孙纪录.非特异性酶对改性几丁质的降解能力及其产物分析[J].中国食品学报,2023,23(4):192-200

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  • 收稿日期:2022-04-23
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  • 在线发布日期: 2023-05-26
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