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CN105838704A 一种纳米纤维生物膜固定化双酶系统及其催化合成海藻糖的方法 无效 - 撤回

一种纳米纤维生物膜固定化双酶系统及其催化合成海藻糖的方法
摘要：
本发明提供了一种纳米纤维生物膜固定化双酶系统及其催化合成海藻糖的方法。本方法在发酵产耐高温海藻糖合酶的大肠杆菌细胞表面自组装淀粉质纳米纤维生物膜，利用可基因编码的多肽标签SpyTag?SpyCatcher特异性共价结合高效固定重组β淀粉酶，自主构建海藻糖合酶—β淀粉酶双酶催化体系，最终使胞外、胞内两步催化连续进行。本方法对β淀粉酶的固定化效率可达到50%?62%；含量为20%?25%重量的可溶性淀粉在胞外经生物膜上的β?淀粉酶催化形成麦芽糖后进入细胞内与海藻糖合成酶反应生成海藻糖，固定化细胞重复利用10?16次后淀粉的转化率可达到45%?55%。
摘要（英）：

The invention provides a nanofiber biological membrane immobilized bi-enzyme system and a trehalose catalytic synthesis method thereof. A starchiness nanofiber biological membrane is arranged on the surface of an escherichia coli cell generating high-temperature-resistant trehalose synthase through fermentation, polypeptide tag SpyTag-SpyCatcher capable of gene coding is used for specificity covalent binding and efficient immobilization recombination of beta-amylase, a trehalose synthase-beta amylase bi-enzyme catalytic system is constructed autonomously, and finally extracellular and intracellular two-step catalysis is conducted continuously. The immobilization efficiency of beta-amylase can reach 50-62%, maltose is generated by 20-25wt% of soluble starch under catalysis of beta-amylase on the extracellular biological membrane, enters the cell and reacts with trehalose synthase to generate trehalose, and the conversion rate of starch can reach 45-55% after the immobilized cell is reused for 10-16 times.

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