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《Life Science Research》 2017-06
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Activity Analysis and Secretory Expression of Human Alpha-defensin 5 in Saccharomyces cerevisiae

SONG Nan;SHAO Jie;LIU Zi-hao;XU Zi-long;XU Qi-yao;ZHANG Xue-wen;CHEN Jin-jun;College of Biological Science and Technology, Hunan Agricultural University;  
The feasibility of secretion expression of human α-defensin 5(HD5), which exhibits the highest antimicrobial activity in human α-defensin family, in Saccharomyces cerevisiae was explored. The DNA fragment containing HD5 coding sequence with biased codons of S. cerevisiae was amplified by PCR, and inserted into the shuttle plasmid pVT102U/α of S. cerevisiae. The recombinant vector named pVT102U/α-HD5 was transformed into S. cerevisiae S78. The positive recombinant clones were screened out under nutrient deficiency. After fermentation, the recombinant protein could be detected in the culture supernatant by tricineSDS-PAGE, and the protein was further subjected to mass spectroscopy to identify the relative molecular mass. The antimicrobial activities of the supernatant against the growth of Escherichia coli, Staphylococcus aureus and Salmonella pullorum were investigated. The minimal concentrations of the lyophilized powder of fermentation supernatant were 10 mg/mL to S. aureus and 40 mg/mL to both E. coli and S. pullorum for complete inhibition. Furthermore, the antibacterial activity of supernatant did not attenuate after treatment with various temperatures up to 140 ℃ for 10 min. The results showed that the recombinant HD5 could be produced with high antibacterial activity.
【Fund】: 湖南省教育厅重点项目(16A098);; 湖南省科技厅重点项目(2017NK2311);; 国家自然科学基金资助项目(31672457 31772865);; 湖南农业大学大学生创新项目(XCX17071)
【CateGory Index】: Q51;Q78
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