近平滑假丝酵母不对称还原同工酶的反应多样性规律及其催化性能调控

The principle of reaction diversity and dissimilarity of isozymes with the same catalytic function from one microbial system is important for control and manipulation of the whole catalytic system and even development of multifunctional biocatalysts. Discovery of the potential dehydrogenase/reductases involved in the objective reaction and elucidation of the relationship between structure and function of the isozymes should be the key issues for understanding the molecular mechanism of reaction diversity among different dehydrogenase/reductases and further function modulation and development of the enzymes from functional microorganism. Based on the genomic sequence of Candida parapsilosis containing stereospecific dehydrogenase/reductases, this research is to search dehydrogenase/reductase isozymes from Candida parapsilosis genome by blastn/p using gene sequence or conserved motif of known functional enzymes as template, to establish the relationship between structure diversity and property diversity and disclose the molecular principle of reaction diversity of stereospecific dehydrogenase/reductases from Candida parapsilosis by detecting the catalytic performance and analyzing the corresponding specific structure, and to enhance the availability of newly discovered stereospecific dehydrogenase/reductases for industrial application by family shuffling and domain swapping of dehydrogenase/reductase isozymes. This research would be favorable for not only understanding the molecular mechanism of reaction diversity of asymmetric reduction-catalyzed isozymes from Candida parapsilosis in theoretical perspectives but also function modulation and availability enhancement of stereospecific dehydrogenase/reductases from the viewpoint of industrial application potential.

微生物催化系统中具有相似功能的同工酶的多样性组成规律及其性能差异的分子基础是整体把握和调控系统效率、开发多功能生物催化剂的前提。如何从功能微生物中发现参与目的反应的潜在脱氢/还原酶同工酶资源，认识同工酶之间反应多样性的结构与性能差异的分子基础，并进一步强化脱氢/还原酶的催化性能，是微生物催化系统中不同立体专一性脱氢/还原酶催化不对称还原反应的规律认识、性能调控和资源开发的关键科学问题。本项目基于近平滑假丝酵母基因组信息，以已知基因序列或保守序列作为模板，在基因组中挖掘潜在新酶资源；通过催化性能表征和特征结构分析，建立结构与性能多样性之间的内在联系，获得近平滑假丝酵母系统中脱氢/还原酶同工酶的反应多样性规律；基于催化性能比较分析，通过同工酶多基因重排、功能结构单元重构等技术，强化其工业属性。基于多样性规律的分子基础，拓展酶资源的高效利用，为不对称合成用新型生物催化剂的开发提供理论和实践基础。

微生物催化系统中具有相似功能的同工酶的多样性组成规律及其性能差异的分子基础是整体把握和调控系统效率、开发多功能生物催化剂的前提。如何从功能微生物中发现参与目的反应的潜在脱氢/还原酶同工酶资源，认识同工酶之间反应多样性的结构与性能差异的分子基础，并进一步强化脱氢/还原酶的催化性能，是微生物催化系统中不同立体专一性脱氢/还原酶催化不对称还原反应的规律认识、性能调控和资源开发的关键科学问题。本项目基于近平滑假丝酵母基因组信息，以已知基因序列或保守序列作为模板，在基因组中挖掘潜在新酶资源；通过催化性能表征和特征结构分析，建立结构与性能多样性之间的内在联系，获得近平滑假丝酵母系统中脱氢/还原酶同工酶的反应多样性规律；基于催化性能比较分析，通过同工酶多基因重排、功能结构单元重构等技术，强化其工业属性。在立体选择性脱氢/还原酶同工酶资源挖掘、分子催化机制解析与突变酶设计、同工酶库构建及其多样性性能评价和酶催化系统的建立等方面取得了相应的研究进展。相关研究成果发表论文20篇，其中SCI论文12篇、CSCD论文8篇；申请中国发明专利4项，授权中国发明专利2项、授权美国发明专利1项；通过中国石油和化学工业联合会组织的科技成果鉴定1项，达到国际领先水平；获得江苏省科学技术奖二等奖1项、中国石油和化学工业联合会技术发明奖一等奖1项。基于多样性规律的分子基础，拓展酶资源的高效利用，为不对称合成用新型生物催化剂的开发提供理论和实践基础。