Purification and characterization of macrolactins and amicoumacins from Bacillus licheniformis BFP011: a new source of food antimicrobial substances

Abstract

In this study, Bacillus licheniformis BFP011 has been cultivated for producing antimicrobial compounds, which were concentrated from the crude supernatant and purified using thin-layer chromatography. The antimicrobial activity was investigated in the presence of several organic solvents and detergents, and inhibiting effects to various Gram-negative and Gram-positive human pathogenic and food spoilage bacteria were observed. Three bands (F4, F5 and F6) showing antimicrobial activity against Salmonella typhi ATCC 5784 were subjected to reversed-phase high-performance liquid chromatography purification. Two peaks of fraction F5, F5-P3 and F5-P4, showed 100% inhibition against S. typhi ATCC 5784. The contained mixture of antimicrobial compounds consists of macrolactins and amicoumacins, which induced the collapse and breaking of S. typhi ATCC 5784 cell membranes in a time-dependent manner. Taken collectively, the results indicate that these compounds may represent promising candidates for the development of food preservative agents of natural origin, as well as novel antimicrobial drug candidates against multiresistant bacterial strains.Purificación y caracterización de macrolactinas y amicoumacinas procedentes de Bacillus licheniformis BFP011: una nueva fuente de sustancias antimicrobianas para los alimentos RESUMEN En el presente estudio, se cultivó Bacillus licheniformis BFP011 para producir compuestos antimicrobianos; estos fueron concentrados del sobrenadante crudo y purificado empleando cromatografía en capa fina (CCF). En presencia de varios solventes y detergentes orgánicos se analizó la actividad antimicrobiana, observándose, además, los efectos inhibidores de varias bacterias gram-negativas y gram-positivas, patógenas para los seres humanos y causantes del deterioro de los alimentos. Mediante RP-HPLC se purificaron tres franjas (F4, F5 y F6) que mostraron actividad antimicrobiana contra la Salmonella typhi ATCC 5784, comprobándose que los dos picos de la fracción F5 (F5-P3 y F5-P4) provocan una inhibición del 100% contra la S. typhi ATCC 5784. La mezcla de compuestos antimicrobianos contiene macrolactinas y amicoumacinas; estas produjeron el colapso y el rompimiento de las membranas celulares de S. typhi ATCC 5784 a medida que fue transcurriendo el tiempo. Considerados en su conjunto, estos resultados indican que es posible que dichos compuestos sean sustancias prometedoras para la creación de agentes conservantes de alimentos de origen natural, y para el desarrollo de medicamentos antimicrobianas novedosas contra cepas bacterianas multirresistentes. ARTICLE HISTORY

Production of Bacteriocin-like Substances by <i>Bacillus</i> Spp. JY-1 in Soy Whey

“…that have a relative narrow inhibitory spectrum [19], the JY-1 supernatant showed a moderate spectrum, including Listeria, Salmonella, Shigella, Staphylococcus, Clostridium and Bacillus, especially L. monocytogenes. Similar results were reported for B. lentus NG121 [20], B. amyloliquefaciens BFP011 [8] and B. licheniformis MCC 2016 [21], but differs from B. amyloliquefaciens EMD17 [5] and B. subtilis KU43 [22], which were not inhibitory to Salmonella.…”

“…Effect of enzymes, pH and heat on the antibacterial activity of Bacillus spp. JY-1 was examined as described by Liu et al [8]. The concentrated supernatant of Bacillus spp.…”

“…Bacillus licheniformis is a gram-positive, spore-forming soli bacterium that is used for the production of enzymes, antibiotics, biochemical and consumer products on an industrial scale [6,7]. B. licheniformis strains isolated from papaya [8] and sediments [9] exhibited a wide spectrum of antimicrobial activity against several bacteria.…”

Classification and Multifaceted Potential of Secondary Metabolites Produced by Bacillus subtilis Group: A Comprehensive Review

“…; S2: Ribosomal peptides (RPs) from B. subtilis group; S3: PolyKetides (PKs) from B. subtilis group; S4: Hybrid (NRP/PK) metabolites from B. subtilis group; S5: Volatile metabolites from B. subtilis group. References [ 209 , 210 , 211 , 212 , 213 , 214 , 215 , 216 , 217 , 218 , 219 , 220 , 221 , 222 , 223 , 224 , 225 , 226 , 227 , 228 , 229 , 230 , 231 , 232 , 233 , 234 , 235 , 236 , 237 , 238 , 239 , 240 , 241 , 242 , 243 , 244 , 245 , 246 , 247 , 248 , 249 , 250 , 251 , 252 , 253 , 254 , 255 , 256 , 257 , …”