Detection of synthetases genes involved in non ribosomallipopeptides (nrlps) biosynthesis from bacillus species by bioinformatics and pcr degenerated primers and estimation of their production. (Funded Research Article)

Abstract

Non Ribosomal Lipopeptides NRLPs produced by Bacillus spp. were firstly detected by bioinformatics in 54 sequenced genomes available from NCBI, which revealed the presence of the four lipopeptides families; surfactin, fengycin/plipastatin, iturin and kurstakin in 19 different strain, fusaricidin from (Paenibacillus) B. polymyxa SC2 and unknown PKS-NRPS lipopeptide from B. safensis FO-36b . Also the absence of NRLPs synthetases genes were detected in 14 different strain, while the 21 other strains harbor different NRP synthetases genes. Secondly, we used the degenerated primers approach to detect the synthetases genes involved in lipopeptides biosynthesis in 9 Bacillus strains. This technique detected non ribosomal synthetases genes for kusrstakin in B. thrunigenesis israelienne NRRL HD-522, surfactin and plipastatin synthetases genes in B. subtilis ATCC 21332 and unknown PKS-NRPS synthetases genes in (Lysinibacillus) B. sphearicus 23268 T. Thirdly, production levels were estimated for these 9 strains by HPLC. The production of B. amyloliquefaciens FZB42 were 294, 62 and 210 mg.L^-1 surfactin, fengycin and mycosubtilin, respectivelly. B. amyloliquefaciens S499 were 872, 103 and 103 mg.L^-1 surfactin, fengycin and bacillomycin respectivelly. B. subtilis ATCC 21332 were 1060 and 226 mg.L^-1 of surfactin and plipastatin respectivelly. While, B. subtilis 168 has no production. The production of B. licheniformis ATCC 14580 strain was 358 mg.L^-1 of lichenysin, and B. pumilus was produced 642 mg.L^-1 of pumilacidin. Also, the strains of B. thuringiensis kurstaki and B. thuringiensis israelienne NRRL HD-522 were produced 102 and 62 mg.g^-1 cells respectivelly of kurstakin.