Immobilization, stabilization and biocatalysis of selected enzymatic systems in non-conventional media

非传统介质中选定酶系统的固定、稳定和生物催化

• 批准号: 36719-2011
• 负责人: Kermasha, Selim
• 金额: $ 1.97万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=568771
• 关键词: Immobilization; stabilization; biocatalysis; selected; enzymatic

The main objective of the applicant's research program is to advance and carry out experimental work that should contribute to the understanding of the mechanistic action and biocatalysis in non-conventional media (NCM), in particular organic solvent media (OSM) and solvent free media (SFM), of selected immobilized enzymes of microbial origin. Although the research work carried out by the applicant addressedthe immobilization and biocatalysis of lipoxygenase/hydroperoxide lyase, the stability of these enzymes as well as the elucidation of their mechanistic action in NCM are still among the major challenges to be overcome. The acquired knowledge and experience in our laboratory in immobilization and biocatalysis of enzymes in non-conventional media led us to move to novel horizons in research, in particular the use of novel nanotechnology approach for enzyme immobilization as well as the modeling and bioinformatics for setting the experimental design for the elucidation of their structure-function in non-conventional media. The specific objectives are: (a) to develop novel non-conventional media (NCM), including organic solvent media (OSM) and free-solvent media (FSM) for higher specificity, catalytic efficiency and stability, using substrate model compounds; (b) to investigate enzyme secondary structure-function relationship in the selected NCM by extensive conformational studies using spectroscopic techniques, including FTIR, circular dichroism and fluorescence for better understanding of their mechanistic actions; (c) to develop novel methodologies, in particular nanoencapsulation, for the immobilization and stabilization of the enzymatic activities of the selected microbial enzymatic extract; and (d) to use the response-surface methodology (RSM) as an approach for investigating the relationship between the catalytic properties of the immobilized enzymes and the nature of supports as well as their effect on internal and external diffusion and on the interfacial surface area or mass transfer.

申请人研究计划的主要目的是进步和开展实验性工作，这些工作应有助于理解非惯性培养基（NCM）的机械作用和生物催化，特别是有机溶剂溶剂介质（OSM）和溶剂溶液的无培养基（SFM）（SFM），对所选的固定型微生物酶的微生物酶。尽管申请人进行的研究工作解决了脂氧酶/氢过氧化物裂解酶的固定和生物催化，但这些酶的稳定性以及其在NCM中的机械作用阐明仍然是要克服的主要挑战之一。我们在非惯性媒体中固定和生物催化的实验室中获得的知识和经验，使我们在研究方面迈向了新的范围，特别是使用新型的纳米技术方法来固定酶以及建模和生物信息学以设定其结构实现媒体的实验设计。 具体目标是：（a）使用底物模型化合物，开发新的非惯性介质（NCM），包括有机溶剂介质（OSM）和自由溶剂介质（FSM），以提高特异性，催化效率和稳定性； （b）使用光谱技术（包括FTIR，圆形二色性和荧光），通过广泛的构象研究来研究所选NCM中的酶二级结构功能关系，以更好地理解其机械作用； （c）开发新的方法，尤其是纳米囊化，以固定和稳定所选微生物酶提取物的酶促活性； （d）使用响应表面方法论（RSM）作为研究固定酶的催化特性与支持性质以及它们对内部和外部扩散以及对种族界面面积或质量转移的影响。