Candida glycerinogenes过量合成甘油过程中渗透压对EMP/HMP途径代谢偏转的调控研究

项目名称： Candida glycerinogenes过量合成甘油过程中渗透压对EMP/HMP途径代谢偏转的调控研究

项目编号： No.31270080

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 诸葛斌

作者单位： 江南大学

项目金额： 78万元

中文摘要： 产甘油假丝酵母（Candida glycerinogenes）是一株能在高渗条件下迅速生长、高效合成甘油、具有自主知识产权的工业菌株，是目前世界合成甘油效率最高的菌株之一。本项目拟利用多年来建立的Candida glycerinogenes研究平台，通过基因敲除、RT-PCR定量分析及产物跟踪分析、二维电泳印迹分析等分子生物学、蛋白组学及代谢组学方法，对高渗条件下EMP/HMP途径关键基因 CgTPI、CgZWF和CgGPD在转录水平及蛋白表达水平上的调控、甘油合成途径中碳代谢及能量代谢的偏转进行研究，从分子水平上揭示该菌株高产甘油的本质，获得耐高渗应答调控机制与甘油过量合成途径的有关新理论。研究结果将为Candida glycerinogenes作为宿主通过代谢工程构建基因工程菌株生产1,3-丙二醇和3-羟基丙酸等三碳高附加值生物产品提供平台和依据。

中文关键词： 产甘油假丝酵母；渗透压；EMP/HMP途径；转录；甘油代谢

英文摘要： Candida glycerinogenes is an industrial strain with independent intellectual property rights that can grow rapidly and synthetize glycerol efficiently in high permeability condition, and it is one of the strains that possess highest synthesis efficiency of glycerol in the word. On the base of research platform built by our laboratory for years, this project aims to investigate the regulation on transcription and protein expression of key genes CgTPI, CgZWF and CgGPD and deflection of carbon and energy metabolism in glycerol synthesis with high permeability condition in EMP/HMP pathway, using molecular biology, proteomics and metabolomics methods such as gene knockout, RT-PCR quantitative analysis and product tracking analysis, and two-dimensional electrophoresis blotting analysis. The essence of high yield of the strain at molecular level will be revealed, and a new theory on the high osmolarity glycerol response mechanism and excessive synthesis of glycerol will be obtained. The results provide a platform for the construction of recombinant bacteria with Candida glycerinogenes as host to produce three carbon substances with high additional values such as 1, 3-propanediol and 3-hydroxyl propionic acid.

英文关键词： Candida glycerinogenes；pressure；EMP/HMP parthway；transcription；glycerol metabolism