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首页> 外文期刊>Journal of the Science of Food and Agriculture >Fusion and secretory expression of an exo-inulinase and ad-allulose 3-epimerase to produced-allulose syrup from inulin
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Fusion and secretory expression of an exo-inulinase and ad-allulose 3-epimerase to produced-allulose syrup from inulin

机译：外氨基蛋白酶和Ad-甲壳酶3-映酶的融合和分泌表达，从菊粉制备甲壳素糖浆

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摘要

BACKGROUND This study developed a feasible catalytic method ford-allulose syrup production using a fusion enzyme, either in free or immobilized form, through hydrolysis of inulin extracted from Jerusalem artichoke tubers. RESULTS d-Allulose 3-epimerase (DAE) was actively expressed in secretory form by fusing with the extracellular exo-inulinase CSCA inEscherichia coliBL21 (DE3). The best linker ligating the two enzymes was a flexible peptide containing 12 residues (GSAGSAAGSGEF). At 55 degrees C and pH 8.0, and as with the addition of 1 mmol L-1Mn2+, the CSCA-linkerE-DAE fusion enzyme obtained through high cell-density cultivation displayed a maximal exo-inulinase activity of 21.8 U mg(-1)and resulted in a yield of 6.3 g L(-1)d-allulose and 39.2 g L(-1)d-fructose using 60 g L(-1)inulin as the raw material. Catechol-modified alginate with titanium ions (Alg(Ti)PDA) was found to be a promising immobilization material for the fusion enzyme. After conversion for 8 days, the Alg(Ti)PDA-immobilized CSCA-linkerE-DAE (8 U g(-1)) completed 24 reaction cycles and retained over 80% of its original activity. Each reaction obtained an average of 19.8 g L(-1)d-allulose and 32.7 g L-1D-fructose from 60 g L(-1)inulin. CONCLUSION This study shed light on a feasible and cost-effective approach for the production of syrup containingd-allulose and D-fructose with inulin as the raw material via the use of a CSCA and DAE fusion enzyme. This syrup is of added value as a functional sweetener. (c) 2020 Society of Chemical Industry
机译：背景本研究开发了一种可行的催化方法，通过水解从菊芋块茎中提取的菊粉，使用游离或固定化形式的融合酶生产菊粉糖浆。结果d-淀粉酶-3-差向异构酶（DAE）与胞外菊粉酶CSCA-inEscherichia coliBL21（DE3）融合，以分泌形式表达。连接这两种酶的最佳连接物是含有12个残基的柔性肽（GSAGSAAGSGGEF）。在55℃和pH 8.0条件下，以及添加1 mmol L-1Mn2+时，通过高细胞密度培养获得的CSCA连接体DAE融合酶显示出21.8 U mg（-1）的最大外链菊糖酶活性，并以60 g L（-1）菊糖为原料获得6.3 g L（-1）d-果糖和39.2 g L（-1）d-果糖。含钛离子的邻苯二酚改性海藻酸钠（Alg（Ti）PDA）是一种很有前途的融合酶固定材料。转化8天后，Alg（Ti）PDA固定化的CSCA-linkerE DAE（8U g（-1））完成了24个反应循环，保留了80%以上的原始活性。每个反应从60 g L（-1）菊糖中平均获得19.8 g L（-1）d-诱惑糖和32.7 g L-1D-果糖。结论本研究为以菊粉为原料，利用CSCA和DAE融合酶生产含有D-果糖和D-果糖的糖浆提供了一种可行且经济的方法。这种糖浆作为功能性甜味剂具有附加值。（c） 2020年化学工业学会

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  • 来源
  • 作者

    Huang Yueyuan; Li Liangfei; Chi Yaowei; Sha Yuanyuan; Wang Rui; Xu Zheng; Xu Xiaoqi; Li Sha; Gao Zhen; Xu Hong;

  • 作者单位

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

    Nanjing Tech Univ State Key Lab Mat Oriented Chem Engn Nanjing Peoples R China;

    Nanjing Tech Univ Coll Food Sci &

    Light Ind Nanjing 211816 Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 农业科学;
  • 关键词

    inulin; fusion enzyme; d-Allulose; Exo-inulinase; immobilized enzyme;

    机译：菊粉;融合酶;d-诱惑;胞外菊粉酶;固定化酶;

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