Research On Reducing The Cell Wall Stability Of Saccharomyces Cerevisiae Using Gene Editing Technology

Saccharomyces cerevisiae is not only rich in various active substances in the cell,but also itself can produce active substances as engineering strain.But the active substances of the yeast cells are surrounded by the thick cell wall.In order to make full use of the abundant material resources in the yeast cells,it is particularly important to study efficient means of breaking the cell wall.The methods of breaking the yeast cell wall mostly focus on chemical,physical and biological methods,but they all have various disadvantages.For example,the chemical method not only destroys the structure of nutrients,but also has a bad impact on the environment;the physical method can cause severe noise pollution;the biological method is relatively expensive and the effects are not very good.At present,the CRISPR/Cas9 system has been used to edit yeast genes and has achieved very significant results.It can be used for mutations in single or multiple genes in the yeast genome,as well as deletion of large fragments of chromosomes and gene insertion.Therefore,in this experiment,CRISPR/Cas9 system was used to explore the effect of the target gene on the stability of the yeast cell wall under specific conditions.This experiment edited the SNF1?DCW1?ROM2?SIT4 genes of S.cerevisiae.The main results were as follows:1.The CRISPR/Cas9 system was used to superimpose editing the SNF1?DCW1?ROM2?SIT4 genes in the genome of S.cerevisiae 2.558 strain,and the mutant strain named 2.558-IV was obtained.2.Exploring the effect of temperature on the cell wall stability of the S.cerevisiae mutant strain 2.558-IV,the experiment showed that the mutant strain was transferred to 38 °C and continued to be cultured for 2 h after 48 h of fermentation,which had a greater impact on the cell wall.The stability of the cell wall was lower,which facilitated the release of active substances.