抗白色念珠菌的海洋来源链霉菌L131的鉴定及活性物质的初步纯化  

Characterization of marine derived Streptomyces sp. L131 with antifungal activity against Candida albicans and preliminary purification of the active fractions

作　　者：王玉梅[1] 董莉[2] 杨天虹[1] 陈德玲[1] 赵心清[1] 

机构地区：[1]大连理工大学生命科学与技术学院,辽宁大连116023 [2]大连理工大学医院,辽宁大连116023

出　　处：《应用海洋学学报》2013年第2期206-214,共9页Journal of Applied Oceanography

基　　金：华东理工大学生物反应器工程国家重点实验室开放课题资助项目;韩国BioGreen 21项目的资助项目(PJ0080932011)

摘　　要：使用卤化酶简并引物对海洋来源的链霉菌进行了基因筛选,共得到了3株阳性菌株,并发现L131和S077菌株具有抗白色念珠菌活性.其中阳性菌株L131的16S rRNA序列与灰白链霉菌(Streptomyces canescens)DSM 40001T具有99%的相似性,但L131菌株在Bennett培养基上菌落凸起,表面褶皱,产白色孢子和淡黄色可溶性色素,不能以阿拉伯糖、果糖作为惟一碳源,与模式菌株存在着明显差异.L131菌株的抗白色念珠菌活性物质在40℃以下,pH值介于5～10范围内活性稳定,且极性大于乙酸乙酯.以AB-8为吸附树脂,首先使用40%(V/V)甲醇洗去未吸附的杂质,然后使用100%(V/V)甲醇进行洗脱,可得到初步纯化的活性物质,其对白色念珠菌的最小抑菌浓度为32.00μg/cm3.上述研究结果可为抗白色念珠菌药物的开发提供参考.New primers were designed based on the conserved sequences of flavin-dependent halogenases, and three positive strains L131 ,S077 and S187 were identified. L131 and S077 showed inhibitory activities against Candida albicans. Analysis of 16S rRNA sequences showed that L131 has the highest similarity（99% ）to S. canescens DSM 40001T. However, L131 distinguished with S. canescens DSM 40001T in culture characters and physio-biochemical features in that L131 has white colonies with ruffled edges, soluble faint yellow pigments on Bennett agar and does not utilizes arabinose and fructose as carbon sources. The culture broth of L131 showed inhibitory activities against C. albicans and the active compounds possessed good stability at temperatures below 40~C in pH from 5 to 10, with stronger polar than ethyl acetate. The antifungal compounds from the crude extract of L131 was absorbed by AB-8 resin and purified further by washing with 40% methanol followed by desorbing using 100% methanol. Thus, the minimum inhibitory concentration of the crude extract was 32.00 μg/cm3. The results presented in this work provides basis for future development of antifungal agents against C. albicans.

关 键 词：微生物学 海洋来源链霉菌 白色念珠菌 抗真菌活性 基因筛选 卤化酶 

分 类 号：Q175.53[生物学—水生生物学]