Author(s): Mesquita, Adriana Spirotto Stein
Date: 2020
Origin: Oasisbr
Subject(s): Espectrometria de massa; Iturina; Metab??litos secund??rios bioativos; Compostos anfif??licos; Atividade surfactante; CI??NCIAS EXATAS E DA TERRA; Lipopept??deos; Bact??rias endof??ticas; Espectrometria de massas; B. amyloliquefaciens
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CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior
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 ??g.mL-1, 58.52 ??g.mL-1 and 66.83 ??g.mL-1 and the LP164 extract showed activity antibacterine against K. pneumoniae with MIC of 62.5 ??g.mL-1.
O g??nero Bacillus possui uma grande variedade de esp??cies produtoras de metab??litos secund??rios bioativos com grande potencial em diferentes campos, entre estes os lipopept??deos, que s??o compostos anfif??licos com atividades antif??ngica, antibi??tica e surfactantes. No presente trabalho foram utilizadas tr??s linhagens de bact??rias endof??ticas de plantas da Amazonia produtoras de lipopept??deos, identificadas por sequenciamento de 16S rDNA como B. amyloliquefaciens. Os dados de espectrometria de massas dos extratos das tr??s linhagens analisadas indicaram a presen??a dos lipopept??deos iturina, surfactina e fengicina. Na avalia????o do rendimento na produ????o de lipopept??deos a partir de diferentes meios de cultivo, o YM apresentou os melhores resultados, tendo a linhagem de B. amyloliquefaciens PpC1 2.2b o maior rendimento com 633,1 mg. L-1. A otimiza????o da produ????o de lipopept??deos para as linhagens de B. amyloliquefaciens PpC1 2.2b, GhcC1 2.1a e DgcR2 1.1b por planejamento fatorial fracionado 24-1 e dados obtidos da ANOVA permitiram propor um modelo eficaz obtendo um aumento significativo da produ????o de lipopept??deos a partir de varia????es de pH, agita????o e aera????o. A partir das an??lises de LC-ESI-MS/MS e RMN dos isolados de PpC1 2.2b foram identificadas dezessete subst??ncias lipopept??dicas incluindo seis hom??logos da iturina A, sete hom??logos da surfactina A, dois hom??logos da fengicina A e dois da fengicina B. As fra????es de surfactinas, iturinas e fengicinas apresentaram atividades anti-inflamat??rias por inibi????o da produ????o de NO em c??lulas de macr??fagos murinos J774A.1 com viabilidade celular acima de 100%. As amostras LP196, Fengicina AB1 e Iturina A2 apresentaram atividade antiplasm??dica contra P. falcipurum FCR3, exibindo CI50 de, 58,52 ??g mL-1, 58,0 ??g.mL-1 e 66,83 ??g.mL-1 e o extrato LP164 apresentou atividade antibacterina contra K. pneumoniae com MIC de 62,5 ??g.mL-1.
