基于半仿生酶解技术的中药乌贼墨最佳用酶初步筛选

Screening of the Best Enzyme for Chinese Squid Ink Based on Semi-bionic Enzymatic Hydrolysis Technique

以体外抗肿瘤实验为指标,初步确定中药乌贼墨的最佳用酶。用人胃肠道系统中的胰蛋白酶,胰凝乳蛋白酶,胃蛋白酶,弹性蛋白酶酶解乌贼墨,以乌贼墨水提液为对照组,在茚三酮显色法测定水解度的基础上,采用四唑盐(MTT)比色法研究四种蛋白酶酶解液对肺腺癌细胞株A549的抑制作用,初步筛选出最佳用酶。四种蛋白酶酶解中药乌贼墨后的蛋白质水解度分别为胰蛋白酶5.12%,胃蛋白酶4.61%,胰凝乳蛋白酶2.09%,弹性蛋白酶0.32%。抗肿瘤实验结果表明:胰蛋白酶酶解液在药物浓度0.4～1.4mg·mL^(-1)范围内,细胞抑制率均在85%～95%。其他三种酶解液的抑制率在各实验浓度范围内均未超过30%。通过半仿生酶解技术,初步筛选出酶解中药乌贼墨的最佳用酶为胰蛋白酶。

In order to determine the optimum enzyme of Chinese traditional medicine cuttlefish ink by anti-tumor experiment in vitro,the squid ink was hydrolyzed by trypsin,chymotrypsin,pepsin and elastase in human gastrointestinal system.And then using the squid ink extract as control group,the hydrolysis degree was determined by ninhydrin chromogenic method.The inhibitory effects of four protease hydrolysates on lung adenocarcinoma cell line A549 were studied by tetrazolium(MTT)colorimetry.The results showed that the degrees of protein hydrolysis of Chinese squid ink by enzymatic hydrolysis of trypsin,pepsin,chymotrypsin and elastase were 5.12%,4.61%,2.09%and 0.32%.The results of antitumor experiment showed that the inhibition rate of trypsin enzymatic hydrolysis solution in the range of 0.4~1.4 mg·mL-1 was in the range of 85%~95%,while the inhibition rates of the other three kinds of enzymatic hydrolysates were not more than 30%in each range of experimental concentration.So the trypsin was screened out as the best enzyme for enzymatic hydrolysis of squid ink.