两种酶水解制备丝素肽的抗菌性及对人胚肾细胞的毒性分析

Antibacterial activity and toxicity to human embryo kidney cells of silk fibroin peptides prepared by two kinds of enzymatic hydrolyses

以废蚕茧为原料,通过Na2CO3脱胶、溶解、透析、冷冻干燥后得到丝素蛋白粉末。用蒸馏水溶解后,分别采用碱性蛋白酶(alcalase)、木瓜蛋白酶(papain)水解丝素蛋白,85℃将酶灭活,得到丝素肽溶液。采用反相高效液相色谱仪对10种丝素肽组分进行分析,并研究丝素肽对大肠杆菌(Escherichia coli)及金黄色葡萄球菌(Staphylococcus aureus)的抗菌效果,以及对人胚肾细胞(HEK293)的毒性。发现P1、A2、A4、A1丝素肽对大肠杆菌生长有抑制作用,P1、P2、P3丝素肽对金黄色葡萄球菌生长有抑制作用,具有抑菌作用的这6种丝素肽对HEK293细胞增殖没有毒性,且A1、A2、A4、P2丝素肽对HEK293细胞存活具有促进作用,为今后丝素肽的产品开发、生产及应用奠定一定的理论基础。

Silk fibroin powder was obtained from waste silkworm cocoon by degumming with Na2 CO3,dissolving,dialysing and freeze-drying. The silk fibroin was hydrolyzed by alcalase and papain in distilled water. The silk fibroin peptides were obtained by inactivating the enzyme at 85 ℃. Reverse phase high performance liquid chromatography(RP-HPLC) was used to analyze the components of the fibroin peptides. Analyze the antimicrobial activity of ten silk fibroin peptides and the cytotoxicity to human embryonic kidney( HEK293) cells. It was found that P1,A2,A4,A1 silk fibroin peptides inhibited Escherichia coli and P1,P2,P3 silk fibroin peptides inhibited Staphylococcus aureus.These six silk fibroin peptides had no toxicity to HEK293 cell proliferation,and A1,A2,A4,P2 silk fibroin peptides could promote the survival of HEK293 cells,laying a theoretical foundation for the development,production and application of silk fibroin peptide in the future.