A Study of Preparation Conditions of PVA Capsule and Immobilization of Biocatalyst

PVA胶囊的制备条件及生物催化剂的固定化研究

• 批准号: 06650910
• 负责人: ARIGA Osamu
• 金额: $ 1.34万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06650910/
• 关键词: polyvinyl alcohol; immobilization; biocatalyst; encapsulation; recombinant; protein; メタノール; β-ガラクトシダーゼ; 包括性

The authors investigated optimal preparation conditions of PVA encapsulation and immobilization of biocatalysts. Toluene-permeabilized cells of Escherichia coli having beta-galactosidase was encapsulated in PVA capsule and kinetic properties of the enzyme were determined assuming Michaelis-Menten and competitive inhibition kinetics. The reduction of Vm is due to inactivation of the enzyme. The Km and Ki of the immobilized enzyme were larger than that of the free enzyme. The increases may have been the contribution by PVA in the core of the capsule, because increases in Km and Ki of the free enzyme by addition of PVA into a reaction mixture was observed. Thus, the immobilization of the enzyme reduced the affinity between the enzyme and substrate. The stability of PVA-encapsulated enzyme was considerably affected by the preparation conditions.The recombinant bacteria were immobilized in PVA capsule and enzyme proteins were produced. This indicates that the proteins can permeate the membrane of a PVA capsule. The productivities of the enzyme were affected by the preparation conditions.

作者研究了 PVA 封装和生物催化剂固定化的最佳制备条件。将具有β-半乳糖苷酶的大肠杆菌的甲苯透化细胞封装在PVA胶囊中，并假设Michaelis-Menten和竞争性抑制动力学测定酶的动力学特性。 Vm 的降低是由于酶的失活。固定化酶的Km和Ki均大于游离酶。这种增加可能是胶囊核心中 PVA 的贡献，因为通过将 PVA 添加到反应混合物中，观察到游离酶的 Km 和 Ki 增加。因此，酶的固定化降低了酶与底物之间的亲和力。 PVA包封酶的稳定性受制备条件影响较大。将重组菌固定在PVA胶囊中，产生酶蛋白。这表明蛋白质可以渗透 PVA 胶囊的膜。酶的生产率受制备条件的影响。