王娅玲　王悦　于佩琳　徐凯旋　陶然　韩金哲　高世杰

摘      要：结合超声法比较酶解前后地胆粉中蛋白质含量的变化，并用正交实验法优化酶解后地胆粉中蛋白质的提取工艺，为今后地胆粉进一步研究提供实验依据。首先，采用胰蛋白酶进行酶解，Lowry法测定酶解前后的蛋白质含量，将酶解与超声法结合，与酶解前进行对比;之后，通过设计不同种类的酶、酶解时间、酶的用量，对地胆粉中蛋白质的提取工艺进行优化。实验表明地胆酶解前的蛋白质含量少于酶解后的蛋白质含量，酶解前采用水提法测定的质量浓度为24.71 mg·mL^-1，胰蛋白酶酶解后蛋白质质量浓度测定为48.32 mg·mL^-1，正交优化地胆的最佳酶解工艺为胰蛋白酶用量为2.5 mg、酶解时间为3.5 h。

关  键  词：地胆;酶解;含量对比;蛋白质;正交实验

中图分类号：TQ 937       文献标识码： A       文章编号： 1671-0460（2020）07-1333-04

Protein Content Determination and Orthogonal Optimization Experiment Before and After Meloecoarctatus Motschulsky Powder Enzymolysis

WANG Ya-ling， WANG Yue， YU Pei-lin， XU Kai-xuan， TAO Ran，，HAN Jin-zhe，GAO Shi-jie

（Shandong University of Traditional Chinese Medicine， Jinan Shandong 250355， China）

Abstract：The change of protein content in Meloecoarctatus Motschulsky powder before and after enzymolysis was compared with ultrasound method， and the extraction process of protein in Meloecoarctatus Motschulsky powder after enzymolysis was optimized by orthogonal experimental method， which could provide experimental basis for further study of Meloecoarctatus Motschulsky powder in the future. At first，the enzymolysis was carried out by trypsin enzymes，and the protein contents before and after the enzymolysis were determined by Lowry method.And then taking enzyme types， enzymolysis time， enzyme dosage as influence factors， the extraction process of protein in Meloecoarctatus Motschulsky powder was optimized，the optimal extraction process was determined. The results showed that the protein content before enzymolysis was less than the protein content after enzymolysis， the content of hydropitimite determination before enzymolysis was 24.71 mg·mL-1， the protein content after enzymolysis was 48.32 mg·mL-1， the best enzymolysis process was as follows： trypsin enzyme 2.5 mg， enzymolysis time 3.5 h.

Key words： Meloecoarctatus Motschulsky powder; Enzymolysis; Content comparison; Protein; Orthogonal experiment

地膽与斑蝥同为芫菁科昆虫，主要成分均为斑蝥素^[1]。目前研究主要集中于斑蝥身上，对其同属动物地胆的研究较少。近年来斑蝥素的抗癌作用备受重视，随着癌症患者数量的增加，对斑蝥素的需求也逐渐上升，导致其供不应求^[2]，因此对于其同属昆虫的开发越来越重要。

地胆为常用动物药，始于《神农本草经》，又名芫青，功用与斑蝥相似，更长于疝积止痛、除疮中恶肉。地胆有大毒，性猛峻裂^[3]，副作用较大，经研究其毒性成分斑蝥素对原发性肝癌、膀胱癌等具有显著抑制作用^[4]，可阻碍肿瘤细胞繁殖，减少不良反应，制备抗肿瘤新药。

酶法为近年来的研究热点，生物酶能破坏细胞壁，加速有效物质溶出，因其专一性而受到广泛欢迎。……