基于超分子结构固定化脂肪酶的构建

项目名称： 基于超分子结构固定化脂肪酶的构建

项目编号： No.20876011

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 谭天伟

作者单位： 北京化工大学

项目金额： 35万元

中文摘要： 基于建立的酶蛋白和载体微环境超分子结构分子模拟和解析方法，构建了形成超分子结构的固定化酶，对依靠大量试验确定固定化方法工作量的减少和成本的降低具有重要意义。课题组研究了酶和微环境超分子结构分子模拟方法、固定化酶超分子结构表征方法、超分子结构固定化酶构建方法，并将其应用于催化体系中进行了调控机理的研究。建立了脂肪酶在溶剂中、与固定化载体纳微结构结合后分子构象变化的分子模拟方法，模拟了在不同有机溶剂、低碳醇水溶液对脂肪酶构象活力变化及葛根素等在C8疏水配基键合poly(GMA-co-EGDMA) 整体柱的分离识别机理。基于圆二色谱法、傅里叶红外等光谱法建立了固定化脂肪酶超分子结构分析表征方法，表征了假丝酵母脂肪酶4种同工酶及超分子配体自组装多肽RATEA16折叠片超分子结构。建立了制备超分子结构载体和表面改性方法，应用于固定化酶，构建了超大孔树脂、氨丙基接枝介孔二氧化硅纳米管及（非）织物载体的改性和固定化，并应用于手性拆分、精细化学品合成研究其催化特性及调控方法。累计发表核心刊物文章22篇，其中SCI论文19篇。申请国家发明专利2项。培养博士4人，硕士6人。获生物炼制会议杰出贡献奖1人。

中文关键词： 分子动力学；超分子；脂肪酶；固定化；调控机理

英文摘要： With molecular dynamics method and supermolecule analysis between enzyme protein and micro-environment, lipase immobilization basing on the enzyme protein and micro-structure was achieved. With this method, the complex supermolecule of immobilized lipase can be determined with lots of reduction in workload and cost, which is important in industry. Supermolecule structure and molecular modeling for lipase microenvironment and the immobilized lipase was investigated in our lab. And the corresponding catalytic mechanism of the supermolecule building immobilized lipase was also studied in varied organic synthesis.Through molecular simulation, the effect of organic solvents on lipase activity and conformation, as well as the recognition mechanism of puerarin and other hydrophobic ligands in the C8-bonded poly (GMA-co-EGDMA) monolithic column in separation was characterized. By circular dichroism, Fourier transform infrared spectroscopy, the structure of four kinds of Candida lipase isozymes and supermolecule self-assembly of peptide ligands RATEA16 pleated sheet supramolecular was characterized. In the study of lipase microenvironment and supermolecule structure, cyclodextrin aqueous and fatty alcohols interaction with lipase revealed the inactivation mechanism of lipase. On-line analysis super-molecular structure from stationary-phase lipase adsorption is important in supermolecule structure formation mechanism. With special CD and FTIR-ATR spectra, immobilized lipase from crude enzyme powder on non-woven fabrics was characterized for the secondary structure. hydrophobic interface of immobilized carrier activation and inactivation of the immobilized enzyme mechanism was verified. Based on supermolecule analysis , macroporous resin, aminopropyl grafted mesoporous silica nanotubes and membrane fabric were used for lipase immobilization, applied in synthesis of racemic ibuprofen, αhenyl ethanol and polyols ester, polyester, etc.

英文关键词： Molecular dynamics；Supermolecule; lipase; immobilization; Regulation mechanism