全蝎白术白头翁组方发酵前后可溶性蛋白质分子量分析
2020-08-23陶然刘富菁王娅玲王悦袁如月宋兴爽王集会
陶然 刘富菁 王娅玲 王悦 袁如月 宋兴爽 王集会



摘 要: 通过Tricine-SDS-PAGE电泳法比较全蝎白术白头翁组方发酵前后水溶性蛋白质相对分子量的变化。按照超声水提的最佳工艺,分别制备全蝎白术白头翁组方(SAPZF)、全蝎白术白头翁组方发酵物(SAPZFFW)的超声水提醇沉冻干粉。通过Tricine-SDS-PAGE电泳法对SAPZF、SAPZFFW醇沉物中的蛋白进行凝胶电泳分离。结果表明:12% T、3% C的Tricine-SDS-PAGE电泳图谱显示SAPZF醇沉物的蛋白主要出现了4个条带,相对分子量主要集中在130、70、50、35 kDa左右;SAPZFFW醇沉物的蛋白主要出现了3个条带,相对分子量分别集中在50、20、15 kDa左右。Tricine-SDS-PAGE电泳图谱显示发酵前后的全蝎白术白头翁中蛋白相对分子量大小具有显著性差别,发酵之后的蛋白质相对分子量较未发酵的变小。
关 键 词:全蝎;白术;白头翁;Tricine-SDS-PAGE电泳;发酵;水溶性蛋白质
中图分类号:R284.1 文献标识码: A 文章编号: 1671-0460(2020)07-1330-04
Analysis on Molecular Weight of Soluble Protein
of SAPZF Before and After Fermentation
TAO Ran a, LIU Fu-jing a, WANG Ya-ling a, WANG Rue a, YUAN-Ruyueb, SONG-Xingshuanga,WANG Ji-hui c
(a.School of Pharmacy;b.College of Traditional Chinese Medicine;c.Experimental Center,
Shandong University of Traditional Chinese Medicine, Jinan Shandong 250355 , China)
Abstract: The change of protein molecular weight of SAPZF lyophilized powder before and after fermentation was compared by Tricine-SDS-PAGE electrophoresis. According to the optimum process of ultrasonic water extraction, the SAPZF and SAPZFFW extract lyophilized powders were prepared, respectively. The alcohol-precipitated proteins in SAPZF, SAPZFFW were separated by Tricine-SDS-PAGE electrophoresis. The results showed that, under the condition of 12% T and 3% C, Tricine-SDS-PAGE electrophoresis map of SAPZF extract showed four bands of protein with 130,70,50,35 kDa; and Tricine-SDS-PAGE electrophoresis map of SAPZFFW extract showed three bands of protein with 50,20,15 kDa. The Tricine-SDS-PAGE electrophoresis map showed that the molecular weight of the protein was significantly different in the extract lyophilized powder of SAPZF and SAPZFFW.
Key words: Scorpio; Atractylodes macrocephala; Pulsatilla chinensis; Tricine-SDS-PAGE electrophoresis; Fermentation; Water soluble protein
全蝎白術白头翁组方(SAPZF)是以全蝎、白术、白头翁三种中药按等量混合组成的复方。前期研究,该组方具有抗肿瘤抗病毒的作用[1],且经发酵后的组方,其抗肿瘤抗病毒作用有明显的提高。球孢白僵菌(B. bassiana)是属于半知菌亚门(deuteromycotina)的昆虫病原真菌,该菌的次生代谢物质如白僵菌素和其他聚酮类物质具有抗癌、抗病毒等作用[2]。相关研究表明[3],大分子蛋白物质能通过固体发酵技术降解为小分子肽类,具有降低中药毒性、提高中药药效的作用。利用球孢白僵菌双向固体发酵技术发酵本复方后,其水溶性蛋白质、水溶性氨基酸的含量升高,抗肿瘤、抗病毒效果增强[4]。考虑其抗肿瘤作用的增强可能与发酵后的蛋白质成分的变化有关,采用 Tricine-SDS-PAGE电泳法对发酵前后全蝎白术白头翁中可溶性蛋白质的种类和相对分子量进行比较,为今后研究其抗肿瘤成分奠定基础。……
