Peng Cheng, An Duopeng, Ding Wen-Xin, Zhu Yuan-Xing, Ye Li, Li Jiyang
Department of Biological Medicines, School of Pharmacy, Fudan University, Shanghai, 201203, China.
Appl Microbiol Biotechnol. 2020 Dec;104(24):10437-10449. doi: 10.1007/s00253-020-10996-z. Epub 2020 Nov 10.
In this study, we isolated an endophytic Streptomyces sp. strain, WP-1, from surface-sterilized barks of Pinus dabeshanensis, an endangered Chinese plant. WP-1 showed strong antifungal activity against diverse pathogenic fungi, such as Fusarium oxysporum, Rhizoctonia solani, Phytophthora infestan, and Candida albicans. Based on phylogenetic analyses, preliminary identification suggested that the WP-1 strain belonged to the genus Streptomyces. Column chromatogram and HPLC were employed to isolate the primary antifungal component from the culture medium of WP-1, and it was identified as the methylpentaene macrolide antibiotic, fungichromin (FC). In this study, for the first time, using in vitro bioassay studies, we revealed that FC strongly inhibited mycelial growth and conidia germination in Fusarium oxysporum. The median inhibitory concentration of FC was found to be 3.80 mg/L. The fermentation conditions of the WP-1 strain were further investigated to improve FC production. We found that supplementation of the synthetic medium with oils (soybean oil, oleic acid, and so on), particularly during the initial stage of fermentation, significantly increased the FC yield. Ammonium-trapping agent (magnesium phosphate) was used as an additive to increase FC yield to 5741.7 mg/L. It was 2.9-fold more as compared to the highest FC yield reported so far where Streptomyces padanus PMS-702 was used for FC production. KEY POINTS: • Isolation and identification of a fungichromin-producing endophytic actinomycete WP-1 strain. • Fungichromin production was significantly improved via oils and ammonium-trapping agents addition. • Discovery of the antifungal activity of fungichromin against Fusarium oxysporum.
在本研究中,我们从中国濒危植物大别山五针松表面消毒的树皮中分离出一株内生链霉菌菌株WP-1。WP-1对多种致病真菌表现出强大的抗真菌活性,如尖孢镰刀菌、立枯丝核菌、致病疫霉和白色念珠菌。基于系统发育分析的初步鉴定表明,WP-1菌株属于链霉菌属。采用柱色谱和高效液相色谱从WP-1的培养基中分离出主要抗真菌成分,经鉴定为甲基戊二烯大环内酯抗生素真菌色素(FC)。在本研究中,我们首次通过体外生物测定研究发现,FC强烈抑制尖孢镰刀菌的菌丝生长和分生孢子萌发。FC的半数抑制浓度为3.80mg/L。进一步研究了WP-1菌株的发酵条件以提高FC产量。我们发现,在合成培养基中添加油类(大豆油、油酸等),尤其是在发酵初期添加,可显著提高FC产量。使用铵捕获剂(磷酸镁)作为添加剂,可将FC产量提高到5741.7mg/L。与迄今为止报道的使用帕氏链霉菌PMS-702生产FC的最高产量相比,该产量提高了2.9倍。要点:• 分离并鉴定了一株产真菌色素的内生放线菌WP-1菌株。• 通过添加油类和铵捕获剂显著提高了真菌色素的产量。• 发现了真菌色素对尖孢镰刀菌的抗真菌活性。