Nanopore Enhanced Lipase Bioreactors

描述(申请人提供)：该项目的主要目标是开发一种平台技术，纳米孔增强酶装置(Need)，用于生产对映体纯(即手性特定的)药物中间体。该酶将固定在纳米孔载体上，其孔形态和表面修饰将专门设计，以进一步提高酶的活性和选择性。本项目的具体目标是：1.纳米孔二氧化硅-壳聚糖复合生物反应器固定化脂肪酶；2.通过修饰纳米孔载体和控制水活度来提高酶的性能；3根据以上三个具体目标，我们建议在本项目过程中实现以下技术里程碑：(1)在保持脂肪酶活性和选择性的同时，将脂肪酶固定在二氧化硅-壳聚糖纳米孔复合材料上；(2)通过工程化的孔结构在纳米复合材料中创建开放的通道结构和高比表面积，显著提高附着的脂肪酶的反应速度；(3)通过在复合材料中引入表面修饰配体来增加疏水-亲水界面面积，以进一步提高脂肪酶的活性；(4)利用壳聚糖相的溶胀比来控制和微调脂肪酶周围的水活性；(5)通过在连续流动反应器中选择性地去除产物来提高反应和分离的效率；(6)在手性特定配体修饰的脂肪酶小球组成的生物反应器中实现同时的手性生产和分离(S)。南极洲假丝酵母脂肪酶B(Calb)将被固定在纳米孔二氧化硅-壳聚糖复合材料上，以取代传统的生产，使用复杂的合成和分离方案，用于制药、食品/营养、LCD行业的精细化学品。

DESCRIPTION (provided by applicant): The main objective of this project is to develop a platform technology, Nanopore Enhanced Enzyme Device (NEED), for the production of enantiomerically pure (i.e. chiral specific) drug intermediates. The enzyme is to be immobilized on a nanopore support whose pore morphology and surface modifications will be specifically designed for further enhancing the enzyme's activity and selectivity. The specific aims of this project are: A.1 Lipase Immobilization with nanopore silica-chitosan composite as a bioreactor A.2 Enhancing Enzyme Performances by modifying nanopore support and controlling water activity A.3 Achieving Reaction and Separation Synergistically in NEED In accordance with the three specific aims set above, we propose to accomplish the following technical milestones during the course of this project: (1) Immobilizing a lipase onto silica-chitosan nanopore composite while maintaining its activity and selectivity; (2) Creating open channel structure and high surface area in the nanocomposite through engineered pore structures to significantly enhance the reaction rate of the attached lipase; (3) Increasing the hydrophobic-hydrophilic interfacial area, by incorporating surface modification ligands, within the composite to further enhance lipase's activity; (4) Using the swelling ratio of the chitosan phase to control and fine tune water activity surrounding the lipase; (5) Enhancing the efficiencies of reaction as well as separation by selectively removing a product in a continuous flow reactor; (6) Accomplishing simultaneous chiral production and separation in a bioreactor composed of lipase beads modified with chiral specific ligand(s). Candida antarctica lipase B (CALB) will be immobilized onto nanopore silica-chitosan composite for possible productions, in replacements of traditional productions using sophisticated synthesis and separation schemes, of fine chemicals to be used by pharmaceutical, food/nutrition, LCD industries.