Genome Analysis And Gene Functional Characterriazation Of Saccharomyces Cerevisiae Rice Wine Strain YHJ7

Rice wine is a traditional brewing food in China.In the process of producing rice wine,Saccharomyces cerevisiae,as the key microorganism,plays a major role in the conversion of sugar into wine.The obtained rice wine has a unique flavor and rich nutrition Substances and higher health benefits.At present,the rice wine yeast not only in the traditional rice wine fermentation has been widely used,but also in the machinery of rice wine production has also achieved good social and economic benefits.In the rice wine production,the fermentation environment has changed,such as temperature,osmotic pressure,pH,ethanol concentration and other changes will have a more significant impact on yeast fermentation growth.Ethanol is one of the main factors restricting the production of alcohol.In this paper,yeast isolated from yellow rice wine fermenting plant preserved in our laboratory was used as a target to further isolate and select a yeast haploid YHJ7(D2)(Saccharomyces cerevisiae),which has good characteristics of high tolerance.The whole genome of this rice strain with excellent resistance to stress was sequenced,assembled and analyzed for its specificity by comparative genomics and bioinformatics.In comparison with the S.cerevisiae N85(Saccharomyces cerevisiae)genome obtained from the same rice wine fermenting plant,a number of possible different fragments were found,and when compared with the model experimental strain S288c,some differences were found,but not present in the yeast Saccharomyces cerevisiae strain S288c(Saccharomyces cerevisiae),such as the new gene g5170.In this paper,the genome assembly and analysis on the basis of the new gene g5170 to do a functional study and validation.Through bioinformatics analysis,our laboratory predicted that the function of the new gene g5170 may have a great relationship with ethanol tolerance.To further confirm that the new gene g5170 and ethanol tolerance,our laboratory use knock:ing out and integration of g5170 gene in the study and verify its function.In this study,we used Cre/loxP knockout gene g5170 to construct the knockout strain of this gene,and compared the gene knockout strain with the original strain.The results showed that the tolerance of the knockout strain YHJ7-Ag5170 to high glucose and high temperature tolerance was almost the same,but the ethanol tolerance of the knockout strain YHJ7-?g5170 was higher than that of the original strain YHJ7 The performance is much weaker.In order to further test the relationship between the new gene g5170 and ethanol tolerance,we used the Saccharomyces cerevisiae expression vector pYES2 to transform the new gene g5170 into the BY4741(Saccharomyces cerevisiae)yeast strain,and induced the expression of the new gene in the BY4741 recombinant strain by galactose induction method.The data show that recombinant yeast strain g5170 is significantly more tolerant to ethanol than the control strain.In conclusion,the function of this gene is related to the adaptation of ethanol yeast to ethanol stress.