摘要
随着真菌感染的发病率及抗药性的逐年上升,筛选新的抗真菌活性物质已成为临床治疗系统性真菌感染的迫切需要。以双层琼脂扩散法测定抗真菌活性,研究10株海洋放线菌的发酵液及其菌体甲醇提取液对白色念珠菌的抑制作用。结果筛选到一株胞内含有抗真菌活性物质的海洋放线菌Actinomycete A9,经10 L发酵罐发酵,将菌体甲醇提取物及发酵液乙酸乙酯萃取液浓缩合并,利用硅胶柱层析、葡聚糖凝胶柱层析Sephadex LH-20等分离纯化手段,最终获得1 mg浅黄色针状晶体的单一化合物,纯度为99.9%。同时,以ConA、LPS诱导的小鼠脾淋巴细胞增殖为模型,发现Actinomycete A9经硅胶柱层析分离后的抗真菌组分(10μg/mL)能够抑制ConA、LPS诱导的小鼠脾淋巴细胞增殖,表明海洋放线菌Actinomycete A9的胞内活性物质具有抗真菌及免疫抑制活性,为后续研究奠定了基础。
The incidence and drug resistance of fungal infection have been increasing in recent years. To meet the urgent need of treating systemic fungal infection, novel antifungal substances have been screened. Antifungal activities of 10 actinomycetes＇ broth and methanol ectract of mycelium were studied by agar punch method. Marine aetinomycete Actinomycete A9 was screened for its intraeellular antifungal product. Fed-batch cultures of Actinomycete A9 were carried out in a 10 L fermentor. The methanol ectract of mycelium and acetic ether extract of broth were concentrated and mixed. The silica gel column chromatography, Sephadex LH-20 and other methods were used to isolate the antifungal substances. 1 mg antifungal pale yellow needle-shaped crystal （ purity 99.9% ） was attained. Meanwhile, antifungal fraction（ 10 txg/mL） separated by silica gel column chromatography could inhibit the spleen lympho- cyte proliferation induced by ConA or LPS. In our study,intracellular antifungal product from marine actinomycete Actinomycete A9 has antifungal and immunosuppressive activity, and these results are useful for further investigation of Actinomyeete A9 in the future. Key words Marine actinomycete, Antifungal activity, Immunosuppressive, Isolation, Column chromatography, HPLC
出处
《药物生物技术》
CAS
2013年第4期326-330,共5页
Pharmaceutical Biotechnology
基金
中国药科大学2012年大学生创新项目
关键词
海洋放线菌
抗真菌活性
免疫抑制
分离纯化
柱层析
HPLC
Marine actinomycete, Antifungal activity, Immunosuppressive, Isolation, Column chromatography, HPLC
