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PCR screening of 3‐amino‐5‐hydroxybenzoic acid synthase gene leads to identification of ansamycins and AHBA‐related antibiotic producers in Actinomycetes
Author(s) -
Huitu Z.,
Linzhuan W.,
Aiming L.,
Guizhi S.,
Feng H.,
Qiuping L.,
Yuzhen W.,
Huanzhang X.,
Qunjie G.,
Yiguang W.
Publication year - 2009
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2008.04010.x
Subject(s) - antibiotics , gene , biology , streptomycetaceae , identification (biology) , biochemistry , genetics , streptomyces , actinomycetales , microbiology and biotechnology , bacteria , botany
Aims: The 3‐amino‐5‐hydroxybenzoic acid (AHBA) synthase is one of the essential and unique enzymes for AHBA biosynthesis. The possibility of screening for ansamycin or AHBA‐related antibiotic‐producing strains from Actinomycetes by targeting an AHBA synthase gene was explored. Methods and Results: A pair of degenerated primers designed according to the conserved regions of five known AHBA synthases was used to detect AHBA synthase genes within the genomic DNA of Actinomycetes. PCR screening resulted in obtaining 33 AHBA synthase gene‐positive strains from 2000 newly isolated Actinomycetes. Phylogenetic analysis of these gene fragments along with those involved in the biosynthesis of structurally determined ansamycins showed that the genes with close phylogenetic relationships might be involved in the biosynthesis of compounds with the same/similar structures. Four strains have been proved to be actual geldanamycin or rifamycin producers by chemical characterization of their fermentation products. Conclusions: The results confirmed the feasibility of using the AHBA synthase gene as a probe in polymerase chain reaction (PCR) screening of ansamycin or AHBA‐related antibiotic‐producing strains. Significance and Impact of the Study: The PCR screening of AHBA synthase gene represents a direct and sensitive molecular method for rapid detection of AHBA‐related antibiotic‐producing strains.