[Rapid identification of elaiophylin from Streptomyces hygroscopicus 17997, a geldanamycin producer].
To identify the anti-bacterial compound(s) from Streptomyces hygroscopicus 17997, a geldanamycin producer, silica gel thin layer chromatography (TLC) TLC was used to separate the secondary metabolites of S. hygroscopicus 17997. Compound(s) from the silica gel TLC with anti-Gram positive bacteria activity and becoming red upon color reaction by 2.0 mol/L NaOH was analyzed by HPLC. The UV absorption profile and the retention time of a peak of HPLC were identical to those of authentic elaiophylin. A conserved region of dTDP-glucose-4,6-dehydratase (Tgd) gene was amplified by PCR from the genomic DNA of Streptomyces hygroscopicus 17997. DNA sequence analysis of the amplified DNA fragment indicated that it should be the tgd gene of elaiophylin biosynthetic gene cluster. These results implied that the compound in the peak of HPLC was elaiophylin, a macrodiolide antibiotic. The compound was then confirmed to be elaiophylin by LC-(+)-ESI-MS, which revealed that Streptomyces hygroscopicus 17997 was an elaiophylin producer. At the same time, a fast procedure, which consisted of silica gel TLC, color reaction, HPLC, PCR detection and DNA sequence analysis of tgd gene, and LC-(+)-ESI-MS, was established for rapid identification of elaiophylin and its producer.
Key Laboratory of Biotechnology of Antibiotics of Ministry of Health, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Shufen
S
Li
Linzhuan
L
Wu
Feifei
F
Chen
Hongyuan
H
Wang
Guizhi
G
Sun
Yiguang
Y
Wang
D016227:Benzoquinones / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D006836:Hydro-Lyases / Q000235:genetics; D047029:Lactams, Macrocyclic / Q000378:metabolism*; D018942:Macrolides / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
D016227:Benzoquinones; D004269:DNA, Bacterial; D047029:Lactams, Macrocyclic; D018942:Macrolides; C033279:elaiophylin; D006836:Hydro-Lyases; C022771:dTDPglucose 4,6-dehydratase; C001277:geldanamycin