Produção e purificação de lipopeptídeos bioativos produzidos por bactérias endofíticas de plantas da região Amazônica

Abstract

The Bacillus genus has a wide variety of species that produce bioactive secondary metabolites with great potential in different fields, including lipopeptides, which are amphiphilic compounds with antifungal, antibiotic and surfactant activities. In the present work, three strains of endophytic bacteria from Amazonian plants producing lipopeptides used, identified by sequencing 16S rDNA as B. amyloliquefaciens. The mass spectrometry data of the extracts of the three strains analyzed indicated the presence of the lipopeptides iturin, surfactin and fengycin. In the evaluation of the yield in the production of lipopeptides from different culture media, YM presented the best results, with the B. amyloliquefaciens PpC12.2b strain having the highest yield, 633.1 mg. L-1. The optimization of lipopeptide production for B. amyloliquefaciens PpC1 2.2b, GhcC1 2.1a and DgcR2 1.1b lines by fractional factorial planning 24-1 and data obtained from ANOVA allowed us to propose an effective model for increasing lipopeptide production from pH variations, agitation and aeration. From the LC-ESI-MS / MS and NMR analyzes of the PpC12.2b isolates, seventeen lipopeptide substances were identified, including six iturin A homologues, seven surfactin A homologues, two fengycin A homologues and two fengycin B. Fractions of surfactins, iturins and fengycin showed anti-inflammatory activities by inhibiting the production of NO in cells of murine macrophages J774A.1 with cell viability above 100%. Samples LP196, Fengycin AB1 and Iturin A2 showed antiplasmodic activity against P. falcipurum FCR3, exhibiting IC50 of 58.0 µg.mL-1, 58.52 µg.mL-1 and 66.83 µg.mL-1 and the LP164 extract showed activity antibacterine against K. pneumoniae with MIC of 62.5 µg.mL-1.