Kanokwiroon Kanyanatt, Teanpaisan Rawee, Wititsuwannakul Dhirayos, Hooper Alan B, Wititsuwannakul Rapepun
Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.
Mycoses. 2008 Jul;51(4):301-7. doi: 10.1111/j.1439-0507.2008.01490.x.
This study aimed at screen for antimicrobial activity present in the non-rubber constituents of rubber latex of Hevea brasiliensis against various microbial strains. An antimicrobial protein, hevein was extracted from the bottom fraction after centrifugation and purified by acetone fractionation and anion exchange chromatography on a DEAE-Sepharose Fast Flow column. This procedure was more efficient and rapid than the previously described procedures. The antimicrobial activity was investigated and revealed that hevein, a small (4.7 kDa) cysteine-rich protein, had strong antimicrobial activity, especially against Candida spp. including Candida albicans, Candida tropicalis and Candida krusei. The MIC80 value for hevein was as low as 12 microg ml(-1) with C. tropicalis ATCC 750. Higher MIC80 values were obtained for C. albicans ATCC 10231 (95 microg ml(-1)) and C. krusei ATCC 6258 (190 microg ml(-1)). To confirm the antifungal activity, hevein also inhibited the growth of those fungi in a disk diffusion assay and its inhibition was enhanced when a Hevea latex protease inhibitor was also included. Hevein at a concentration of 30 microg ml(-1) also caused a Ca2+-dependent aggregation of C. tropicalis yeast cells. These data indicate that hevein can inhibit the growth of certain potential oral fungal pathogens.
本研究旨在筛选巴西橡胶树胶乳非橡胶成分中存在的针对各种微生物菌株的抗菌活性。一种抗菌蛋白——橡胶素,从离心后的底部组分中提取,并通过丙酮分级分离和在DEAE-琼脂糖快速流动柱上的阴离子交换色谱法进行纯化。该方法比先前描述的方法更高效、快速。对抗菌活性进行了研究,结果表明,橡胶素是一种小(4.7 kDa)的富含半胱氨酸的蛋白质,具有很强的抗菌活性,尤其是对包括白色念珠菌、热带念珠菌和克柔念珠菌在内的念珠菌属。橡胶素对热带念珠菌ATCC 750的MIC80值低至12微克/毫升。白色念珠菌ATCC 10231(95微克/毫升)和克柔念珠菌ATCC 6258(190微克/毫升)的MIC80值更高。为了证实抗真菌活性,橡胶素在纸片扩散试验中也抑制了这些真菌的生长,并且当加入巴西橡胶树胶乳蛋白酶抑制剂时其抑制作用增强。浓度为30微克/毫升的橡胶素还导致热带念珠菌酵母细胞发生钙离子依赖性聚集。这些数据表明,橡胶素可以抑制某些潜在的口腔真菌病原体的生长。