Expression of chitosanase from Aspergillus fumigatus chitosanase in Saccharomyces cerevisiae by CRISPR-Cas9 tools  

作　　者：Qingshuai Zhang Hui Cao 

机构地区：[1]Beijing Key Lab of Bioprocess,College of Life Science and Technology,Beijing University of Chemical Technology,Beijing 100029,China

出　　处：《Bioresources and Bioprocessing》2024年第1期273-280,共8页生物资源与生物加工(英文)

基　　金：supported by the National Key Research and Development Program of China(2021YFF0600703).

摘　　要：Chitooligosaccharides(COS)find numerous applications due to their exceptional properties.Enzymatic hydrolysis of chitosan by chitosanase is considered an advantageous route for COS production.Heterologous expression of chitosanase holds significant promise,yet studies using commonly employed Escherichia coli and Pichia pasto-ris strains encounter challenges in subsequent handling and industrial scalability.In this investigation,we opted for using the safe yeast strain Saccharomyces cerevisiae(GRAS),obviating the need for methanol induction,resulting in successful expression.Ultimately,utilizing the GTR-CRISPR editing system,shake flask enzyme activity reached 2 U/ml.The optimal chitosanase activity was achieved at 55℃and pH 5,with favorable stability between 30 and 50°C.Following a 2-h catalytic reaction,the product primarily consisted of chitobiose to chitotetraose,predominantly at the chitotriose position,with a slight increase in chitobiose content observed during the later stages of enzymatic hydrolysis.The results affirm the feasibility of heterologous chitosanase expression through Saccharomyces cerevisiae,underscoring its significant industrial potential.

关 键 词：CHITOSAN CHITOSANASE Saccharomyces cerevisiae GTR-CRISPR 

分 类 号：Q55[生物学—生物化学]