Rapid detection and characterization of surfactin-producing Bacillus subtilis and closely related species based on PCR.
The surfactin production genetic locus ( sfp) is responsible for the ability of Bacillus subtilis to produce the lipopeptide biosurfactant, surfactin. This report demonstrates the utility of using PCR of the sfp gene as a means of identifying Bacillus species that produce surfactin. We carried out a hemolysis zone assay, quantitative HPLC and NMR in parallel to ensure that the PCR provided correct results. PCR analyses were performed for the sfp gene on 15 standard strains and 20 field-collected Bacillus spp. isolates native to Taiwan. Among the 15 standard strains, surfactin was produced by seven strains of B. subtilis and two closely related species, B. amyloliquefaciens B128 and B. circulans ATCC 4513. Of the 20 field-collected Bacillus spp. isolates; 16 strains yielded surfactin- positive results with PCR and HPLC. A good correlation was observed. Within the 16 field isolates, B. amyloliquefaciens S13 (452.5 mg/L) and B. subtilis S15 (125.6 mg/L) had high productivity of surfactin. The technique is valuable for finding out potential good yields of surfactin-producing strains. The PCR method we used could also be used to find different species or genera containing homologous genes. This is the first report of the detection of surfactin production by B. amyloliquefaciens and B. circulans based on PCR screening.
Hsieh
Feng-Chia
FC
Biopesticide Division, Taiwan Agricultural Chemicals and Toxic Substances Research Institute, Council of Agriculture, 11 Kuang-Ming Road, Wufeng, Taichung Hsien 413, Taiwan.
Li
Mei-Chen
MC
Lin
Tsung-Chun
TC
Kao
Suey-Sheng
SS
D001407:Bacillus / Q000235:genetics / Q000378:metabolism; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D055666:Lipopeptides; D010453:Peptide Synthases / Q000235:genetics; D010456:Peptides, Cyclic / Q000096:biosynthesis*; D016133:Polymerase Chain Reaction; D013045:Species Specificity
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D055666:Lipopeptides; D010456:Peptides, Cyclic; C009365:surfactin peptide; D010453:Peptide Synthases; C069542:surfactin synthetase