Identification of antifungal niphimycin from Streptomyces sp. KP6107 by screening based on adenylate kinase assay.
Microbial culture extracts are used for natural product screening to find antifungal lead compounds. A microbial culture extract library was constructed using 343 actinomycete isolates to examine the value of the adenylate kinase (AK) assay for screening to identify antifungal metabolites that disrupt cell integrity in plant pathogenic fungi. A culture extract of Streptomyces sp. strain KP6107 lysed cells of Fusarium oxysporum f.sp. lycopersici which resulted in high AK activity. The active ingredient N-1 was purified from the culture extract using various chromatographic procedures and identified to be the guanidyl-polyol macrolide antibiotic, niphimycin, which is a potent fungal cell membrane disruptor. Niphimycin showed broad-spectrum antifungal activity against Alternaria mali, Aspergillus oryzae, Colletotrichum coccodes, Colletotrichum gloeosporioides, Cercospora canescens, Cylindrocarpon destructans, F. oxysporum f.sp. cucumerinum, F. oxysporum f.sp. lycopersici, and Rhizoctonia solani at concentrations of 8-64 µg ml(-1). Anthracnose development in pepper plants was completely inhibited by treatment with 50 µg ml(-1) niphimycin, which was as effective as chlorothalonil. These results show that the AK assay is an efficient and selective tool in screening for cell membrane/wall disruptors of plant pathogenic fungi.
Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea.
Hye Yoon
HY
Kim
Jeong Do
JD
Kim
Jin Sung
JS
Hong
Jong Hyun
JH
Ham
Beom Seok
BS
Kim
D000263:Adenylate Kinase / Q000737:chemistry*; D001203:Ascomycota / Q000254:growth & development; D001681:Biological Assay; D020231:Colletotrichum / Q000254:growth & development; D005659:Fungicides, Industrial / Q000737:chemistry* / Q000302:isolation & purification; D005670:Fusarium / Q000254:growth & development; D006146:Guanidines / Q000737:chemistry / Q000302:isolation & purification; D008826:Microbial Sensitivity Tests; D010944:Plants / Q000382:microbiology; D013302:Streptomyces / Q000254:growth & development
D005659:Fungicides, Industrial; D006146:Guanidines; C010492:niphimycin; D000263:Adenylate Kinase