Design of fine particles of silicon polymer highly effective as entrapping gel for biocatalysts functioning in organic media

硅聚合物细颗粒的设计可作为有机介质中生物催化剂的高效捕获凝胶

• 批准号: 07650960
• 负责人: KAWAKAMI Koei
• 金额: $ 1.34万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07650960/
• 关键词: immobilized enzyme; gel-entrapment; lipase; thermostability; ester synthesis; microemulsion; sol-gel method; organic-inorganic hybrid gel; 熱安定性; 固定化窒素

Entrapment of lipase in organically modified silicates, which are derved from a mixture of tetramethoxysilane (TMOS) and alkyl group-containing organic silanes by the sol-gel method, leads to enhancement in esterification activity and thermal stability of immobilized preparation. In this study, we tried to form the lipase-entrapping gels either (1) in W/O microemulsion or (2) on the surface of support materials, and investigated some enzymatic properties of resultant immobilized lipase. In ester synthesis from butyric acid either isoamyl alcohol or ((]SY.+-。[))-menthol as a test reaction, the following results were obtained.(1) Microemulsion-based entrapment : Among various surfactants tested, the silicate-entrapped lipase prepared by using Brij 97 displayd a highest catalytic activity at room temperature. The organic-inorganic hybrid silicate derived from a mixture of TMOS and methyltrimethoxysilane resulted in a more enhanced activity of the immobilized lipase.(2) Dispersed entrapmen … More t on support surface : Among various organic silanes mixed with TMOS,alkyltrimethoxysilanes bearing longer chain length than propyl group, and methoxysilanes with multiple methyl groups brought about enhanced activity and thermostability of the immobilized lipase preparations. When kieselguhr such as commercially available Celite 545 was used as a support material, the entrapped lipase gave a mzximum esterification activity. There existed an optimum amount of gel and an optimum molar ratio of organic silane to TMOS.As the result of optimization of various preparation conditions, the thermostability of entrapped lipase was enhanced by 20-30ﾟC as compared with that of deposited counterpart.These results and findings might be ascribed to 1) stabilization of tertiary structure of protein globules by reasonably tight entrapment in close proximity of the hybrid gel surface, 2) preferential partitioning of substrates to the hydrophobic gel, and 3) hydrophobic interaction between lipase molecules and alkyl groups of the organic silica gel. Less

以四甲氧基硅烷（TMOS）和含烷基的有机硅烷为原料，采用溶胶-凝胶法制备了有机改性硅酸盐，将脂肪酶包埋在其中，可提高固定化产物的酯化活性和热稳定性。本研究尝试在W/O型微乳液中和载体材料表面制备固定化脂肪酶的凝胶，并对所得固定化脂肪酶的酶学性质进行研究。在由丁酸合成酯的过程中，可以使用异戊醇或（（）SY.+-。[10]）-薄荷醇作为测试反应，获得以下结果。(1)微乳液包埋法：在所测试的各种表面活性剂中，Brij 97制备的硅酸酯包埋脂肪酶在室温下表现出最高的催化活性。由TMOS和甲基三甲氧基硅烷的混合物衍生的有机-无机杂化硅酸盐导致更增强的固定化脂肪酶的活性。(2)分散诱捕 ...更多信息 在支撑表面上测试：在与TMOS混合的各种有机硅烷中，具有比丙基更长的链长的烷基三甲氧基硅烷和具有多个甲基的甲氧基硅烷带来了固定化脂肪酶制剂的活性和热稳定性的增强。当硅藻土如市售的Celite 545用作载体材料时，包埋的脂肪酶具有最大的酯化活性。存在一个最佳的凝胶用量和有机硅烷与TMOS的最佳摩尔比，通过对各种制备条件的优化，包埋脂肪酶的热稳定性比沉淀脂肪酶提高了20-30 ℃，这些结果和发现可能归因于：通过合理紧密地包埋在杂化凝胶表面附近来稳定蛋白质球的三级结构，2）底物优先分配到疏水凝胶，和3）脂肪酶分子和有机硅胶的烷基之间的疏水相互作用。少

项目成果

K.Kawakami: "Enhancement of Thermostability of Lipase by the Sol-Gel Entrapment into Methyl-Substituted Organic Silicates Formed on Diatomaceous Earth" Biotechnology Techniques. 10,7. 491-494 (1996)

K.Kawakami：“通过溶胶-凝胶捕获硅藻土上形成的甲基取代的有机硅酸盐来增强脂肪酶的热稳定性”生物技术。

K.Kawakami: "Immobilization of Lipase into Fine Particles of Silica Gel by the Sol-Gel Transition in Water-in-Oil Microemulsions" Bulletin of Faculty of Engineering, Kyushu Sangyo University. 33. (1997)

K.Kawakami：“通过油包水微乳液中的溶胶-凝胶转变将脂肪酶固定在硅胶细颗粒中”九州产业大学工学部通报。

K.Kawakami: "Immobilization of Lipase into Fine Particles of Silica Gel by the Sol-Gel Transition in Water-in-Oil Microemulsions" Bull.Facul.Eng.,Kyushu Sangyo Univ.33. (1997)

K.Kawakami：“通过油包水微乳液中的溶胶-凝胶转变将脂肪酶固定到硅胶细颗粒中”Bull.Facul.Eng.，九州产业大学33。

K.Kawakami: "Sol-Gel Entrapment of Lipase Using a Mixture of Tetramethoxysilane and Methyltrimethoxysilane as the Alkoxide Precursor : Esterification Activity in Organic Media" Biotechnology Techniques. 9-10. 701-704 (1995)

K.Kawakami：“使用四甲氧基硅烷和甲基三甲氧基硅烷的混合物作为醇盐前体的脂肪酶溶胶-凝胶捕获：有机介质中的酯化活性”生物技术。

K.Kawakami: "Sol-Gel Entrapment of Lipase Using a Mixture of Tetramethoxysilane and Methyl-trimethoxysilane as the Alkoxide Precursor : Esterification Activity in Organic Media" Biotechnology Techniques. 9. 701-704 (1995)

K.Kawakami：“使用四甲氧基硅烷和甲基三甲氧基硅烷的混合物作为醇盐前体的脂肪酶溶胶-凝胶捕获：有机介质中的酯化活性”生物技术。