Under Optimization Of MAPase Of Zygosaccharomyces Rouxii And Salt-tolerance Mechanism Study

Salt-tolerance Zygosaccharomyces rouxii is one of the main strains which can form fermented flavor in later period of Soy sauce. In order to study its salt tolerance mechanism, this study uses the Zygosaccharomyces rouxii which is main aroma producing yeast in soy sauce as the research object, after optimization of Zygosaccharomyces rouxii producing MAP kinase conditions through response surface method, and studying its salt tolerance mechanism through the proteomics. Firstly, explore the preparation of the sample of Zygosaccharomyces rouxii, then extract different protein spot by two-dimensional electrophoresis, and select6spots have the mass spectrum detection, conclusions are drawn:1.Through three single test factors such as temperature, time, concentration to determine the conditions that the best single factor is at temperature of28℃, time is3D, salt concentration is12%, through the response surface method finding that the optimal MAPK conditions were as follows:the temperature was27℃, salt concentration9.2%, incubation time80.4hours. Under this condition the MAP kinase’s concentration was3.189ng/mL.2.In order to finding the most suitable method to extract the protein of Zygosaccharomyces rouxii,the study adopts three methods to extract the protein, such as the repeated freeze-thaw method, liquid nitrogen grinding method and ice bath ultrasonic method. Compared with the effects above, I found that the optimal extract was ice bath ultrasonic method, the total time is15min, and whenever ultrasound3s, stop3s.3. Through separation of the normal group and the high-salt group of proteins by two-dimensional electrophoresis technology, as well as by ImageMaster2D Platinum V5software to analysis, then we get that there are63spots difference expressed more than1.5times, of which there are29increased, and34decreased, and there are10spots expressed more than3times, of which there are5points increased, and5point decreased.4. Through MALDI-TOF,6different proteins (spots) were identified. These proteins in Zygosaccharomyces rouxii under salt stress participated in the following metabolic process, energy metabolism, carbohydrate metabolism, and cell signal transduction pathways.