蚕蛹虫草生物活性肽的制备及其性能分析  

Preparation and property analysis of polypeptides in Cordyceps militaris

作　　者：郭尚 徐莉娜 李艳婷 郭伟伟 郭霄飞 洪沙沙 GUO Shang;XU Lina;LI Yanting;GUO Weiwei;GUO Xiaofei;HONG Shasha(Shanxi Institute for Functional Food,Shanxi Agricultural University,Taiyuan 030031,Shanxi,China)

机构地区：[1]山西农业大学、山西功能食品研究院,山西太原030031

出　　处：《精细化工》2022年第12期2481-2491,共11页Fine Chemicals

基　　金：山西省青年科学研究项目(202103021223160);山西农业大学博士科研启动项目(2021BQ45);山西省来晋奖励博士科研启动项目(SXBYKY2021086);山西省农业科学院科研创新团队专项(YGC2019TD03)。

摘　　要：以蚕蛹虫草子实体干粉为材料,采用碱溶酸沉法提取蚕蛹虫草蛋白质,通过正交实验优化提取条件;通过蛋白质复合酶酶解蚕蛹虫草蛋白质,超滤法分离获得蚕蛹虫草小分子多肽液;通过测定抑菌圈和细胞毒性实验评价蚕蛹虫草多肽的抗菌及抗癌活性。结果显示,蚕蛹虫草蛋白质最佳提取条件为:pH 8.5、料液比(g∶m L)为1∶28、提取时间3.5 h、提取3次,蚕蛹虫草蛋白质含量最高为45.06%;蚕蛹虫草多肽最佳酶解工艺为:碱性蛋白酶与木瓜蛋白酶酶活力比4∶3、酶解最佳p H 7.2、温度55℃、酶加量7000 U/m L、酶解时间为3.5 h。在此条件下制备的蚕蛹虫草多肽含量为16.73%,其相对分子质量<3.0×10^(3)Da组分对大肠杆菌、枯草芽孢杆菌及金黄色葡萄球菌均表现出较好的抑菌效果,其抑菌圈分别为(12.08±0.22)、(6.67±0.12)和(10.32±0.23)mm;对骨肉瘤Sao-S细胞[半数抑制浓度(IC_(50))为0.49 mg/L]和膀胱癌T24细胞(IC_(50)为0.23 mg/L)有很好的抑制作用。Cordyceps militaris protein was firstly extracted from Cordyceps militaris fruiting body dry powder by alkali solution and acid precipitation method,of which the conditions were optimized by orthogonal experiment.Cordyceps militaris protein was then digested by protein complex enzyme,and small molecule polypeptide solution was obtained by ultrafiltration.The antibacterial and anticancer performances of Cordyceps militaris polypeptide were further evaluated via bacteriostatic circle measurement and cytotoxicity test.The results showed that the protein content could reach 45.06%under optimized extraction conditions of pH 8.5,material-liquid ratio(g∶m L)1∶28,and extraction time 3.5 h with the solution being extracted three times.Meanwhile,the optimum enzymatic hydrolysis process of Cordyceps militaris polypeptide was as follows:enzyme activity of alkaline protease to papain 4∶3,enzymolysis temperature 55℃,pH 7.2,enzyme dosage 7000 U/mL,and enzymolysis time 3.5 h.Under these conditions,the prepared polypeptides from Cordyceps militaris had 16.73%content,and that with relative molecule mass<3.0×10^(3)Da generated bacteriostatic circles of(12.08±0.22),(6.67±0.12)and(10.32±0.23)mm against Escherichia coli,Bacillus subtilis and Staphylococcus aureus,respectively,showed good antibacterial activity.Moreover,the osteosarcoma SAO-S[median inhibition concentration(IC_(50))=0.49 mg/L]and human bladder cancer cells(IC_(50)=0.23 mg/L)were significantly inhibited by Cordyceps militaris polypeptide.

关 键 词：蚕蛹虫草 酶解 多肽 抗菌性 抗癌作用 中药现代化技术 

分 类 号：R284.2[医药卫生—中药学]