Preparation of Industrial Hemp Seed Polypeptide by a Whole-cell Biocatalyst Displayed NX1101 Protease on the Surface of Saccharomyces cerevisiae
The preparation of industrial hemp seed polypeptide by enzymatic hydrolysis has the ad-vantages of good safety,low energy consumption and little nutritional loss.In this study,based on the synthesis of defatted hemp seed meal protein extraction and proteolytic enzyme screening,the shuttle plasmid pYD1 was used as the expression vector to anchor proteolytic enzyme NX1101 from bovine intes-tinal metgenomic library on the surface of Saccharomyces cerevillus(EYB100),and the recombinant strain was used as the whole cell catalyst to prepare hemp polypeptides.The enzymatic properties analysis of the whole cell catalyst showed that the most suitable reaction pH value was 8.0,and the enzyme activi-ty stability was high between 8 and 10,so it was an alkaline resistant protease.The optimum reaction temperature was 50℃,and the stability of enzyme activity was higher below 50℃.A total of 1095 pep-tides were obtained by enzymolysis of the defatted hemp protein with the whole cell catalyst of recombi-nant yeast displaying NX1101,among which 140 short peptides with amino acid number≤10 were ob-tained.The enzymolysis efficiency of whole cell catalyst was more than 6.5 times higher than that of con-ventional enzymolysis technology with alkaline protease powder.
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