Metabolic Profiling and Comparative Proteomic Insight in Respect of Amidases during Iprodione Biodegradation.

The fungicide iprodione (IPR) (3-(3,5-dichlorophenyl) N-isopropyl-2,4-dioxoimidazolidine-1-carboxamide) is a highly toxic compound. Although IPR has been restricted, it is still being applied in many places around the world, constituting an environmental risk. The biodegradation of IPR is an attractive option for reducing its residues. In this study, we isolated thirteen IPR-tolerant bacteria from a biopurification system designed to treat pesticides. A study of biodegradation using different strains was comparatively evaluated, and the best degradation rate of IPR was presented by Achromobacter sp. C1 with a half-life (T1/2) of 9 days. Based on a nano-LC-MS/MS analysis for the strains, proteins solely expressed in the IPR treatment were identified by highlighting the strain Achromobacter sp. C1, with 445 proteins primarily involved in the biosynthesis of secondary metabolites and microbial metabolism in diverse environments. Differentially expressed protein amidases were involved in six metabolic pathways. Interestingly, formamidase was inhibited while other cyclases, i.e., amidase and mandelamide hydrolase, were overexpressed, thereby minimizing the effect of IPR on the metabolism of strain C1. The dynamic changes in the protein profiles of bacteria that degrade IPR have been poorly studied; therefore, our results offer new insight into the metabolism of IPR-degrading microorganisms, with special attention paid to amidases.

Biosurfactant Production by Bacillus amyloliquefaciens C11 and Streptomyces lavendulae C27 Isolated from a Biopurification System for Environmental Applications.

Biosurfactant-producing bacteria can be found in contaminated environments such as biopurification systems (BPS) for pesticide treatments. A total of 18 isolates were screened to determine their ability to produce extracellular biosurfactants, using olive oil as the main carbon source. Out of the eighteen isolates, two strains (C11 and C27) were selected for biosurfactant production. The emulsification activities of the C11 and C27 strains using sunflower oil was 58.4 and 53.7%, respectively, and 46.6 and 48.0% using olive oil. Using molecular techniques and MALDI-TOF, the strains were identified as Bacillus amyloliquefaciens (C11) and Streptomyces lavendulae (C27). The submerged cultivation of the two selected strains was carried out in a 1 L stirred-tank bioreactor. The maximum biosurfactant production, indicated by the lowest surface tension measurement, was similar (46 and 45 mN/m) for both strains, independent of the fact that the biomass of the B. amyloliquefaciens C11 strain was 50% lower than the biomass of the S. lavendulae C27 strain. The partially purified biosurfactants produced by B. amyloliquefaciens C11 and S. lavendulae C27 were characterized as a lipopeptide and a glycolipid, respectively. These outcomes highlight the potential of the selected biosurfactant-producing microorganisms for improving pesticides' bioavailability and therefore the degradational efficacy of BPS.

Pesticide-tolerant bacteria isolated from a biopurification system to remove commonly used pesticides to protect water resources.

In this study, we selected and characterized different pesticide-tolerant bacteria isolated from a biomixture of a biopurification system that had received continuous applications of a pesticides mixture. The amplicon analysis of biomixture reported that the phyla Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria were predominant. Six strains grew in the presence of chlorpyrifos and iprodione. Biochemical characterization showed that all isolates were positive for esterase, acid phosphatase, among others, and they were identified as Pseudomonas, Rhodococcus and Achromobacter based on molecular and proteomic analysis. Bacterial growth decreased as both pesticide concentrations increased from 10 to 100 mg L-1 in liquid culture. The Achromobacter sp. strain C1 showed the best chlorpyrifos removal rate of 0.072-0.147 d-1 a half-life of 4.7-9.7 d and a maximum metabolite concentration of 2.10 mg L-1 at 120 h. On the other hand, Pseudomonas sp. strain C9 showed the highest iprodione removal rate of 0.100-0.193 d-1 a half-life of 4-7 d and maximum metabolite concentration of 0.95 mg L-1 at 48 h. The Achromobacter and Pseudomonas strains showed a good potential as chlorpyrifos and iprodione-degrading bacteria.

Antarctic Streptomyces fildesensis So13.3 strain as a promising source for antimicrobials discovery.

Antarctic have been suggested as an attractive source for antibiotics discovery and members of Streptomyces genus have historically been studied as natural producers of antimicrobial metabolites. Nonetheless, our knowledge on antibiotic-producing Streptomyces from Antarctic is very limited. In this study, the antimicrobial activity of organic extracts from Antarctic Streptomyces strains was evaluated by disk diffusion assays and minimum inhibitory concentration. The strain Streptomyces sp. So13.3 showed the greatest antibiotic activity (MIC = 15.6 µg/mL) against Gram-positive bacteria and growth reduction of Gram‒negative pathogens. The bioactive fraction in the crude extract was revealed by TLC‒bioautography at Rf = 0.78 with molecular weight between 148 and 624 m/z detected by LC-ESI-MS/MS. The strain So13.3 was taxonomically affiliated as Streptomyces fildesensis. Whole genome sequencing and analysis suggested a 9.47 Mb genome size with 42 predicted biosynthetic gene clusters (BGCs) and 56 putative clusters representing a 22% of total genome content. Interestingly, a large number of them (11 of 42 BGCs and 40 of 56 putative BGCs), did not show similarities with other known BGCs. Our results highlight the potential of the Antarctic Streptomyces strains as a promising source of novel antimicrobials, particularly the strain Streptomyces fildesensis So13.3, which first draft genome is reported in this work.

Streptomyces luridus So3.2 from Antarctic soil as a novel producer of compounds with bioemulsification potential.

The present study aimed to identify novel microbial producers of bioemulsificant compounds from Antarctic soils. Fifty-nine microbial strains were isolated from five different locations at South Shetland Islands, Antarctica, and screened for biosurfactant production by ß-hemolytic activity. Strain So 3.2 was determined as bioemulsifier-producer and identified by phenotypic and molecular characterization as Streptomyces luridus. Emulsification activity, oil displacement method and drop-collapsing test were performed to evaluate the biosurfactant activity with different oils and hydrocarbons using two different culture media (Luria Bertani and Bushnell Haas in the presence of different carbon sources: glucose, glycerol, olive oil and n-Hexadecane). Cell free supernatant of Bushnell Haas culture supplemented with n-Hexadecane showed the best results for all tests. Emulsification of hydrocarbons exceeded 60%, reaching up to 90% on oil with high API grade, while displacement tests ranged from 8 cm to 4 cm in diameter according the culture media and tested oils. Our results revealed that Streptomyces luridus So3.2 is able to produce bioemulsifiers capable of emulsifying hydrocarbons and oils, which could be used in different biotechnological applications, particularly for bioremediation of environments contaminated by oil leaks.

Nanopartículas Sintetizadas por Bacterias Antárticas y sus Posibles Mecanismos de Síntesis / Nanoparticles Synthesized by Antarctic Bacteria and their Possible Mechanisms of Synthesis

En los últimos años microorganismos tales como hongos, levaduras y, en especial, las bacterias han sido utilizadas para realizar biosíntesis de nanopartículas. Existen varios tipos de bacterias descritas como productoras de nanopartículas, sin embargo, las bacterias psicrófilas y psicrotolerantes no han sido ampliamente estudiadas, aun cuando su utilización en la producción de nanopartículas podría entregar ventajas relacionadas con su estabilidad, el gasto energético de su producción, al mismo tiempo que son una alternativa amigable con el medio ambiente. Este artículo entrega una breve revisión de las bacterias antárticas psicrófilas y psicrotolerantes sintetizadoras de nanopartículas, los posibles mecanismos que se asocian a esta síntesis y perspectivas futuras relacionadas a la biosíntesis bacteriana de nanopartículas.

In recent years microorganisms as fungi, yeasts and especially bacteria have been used to produce nanoparticles biosynthesis. Several types of bacteria are described as nanoparticles producers, however, psychrophilic and psychrotolerant bacterias have not been studied widely, although its use in the production of nanoparticles could provide advantages related to the stability of nanoparticles, energy expenditure on its production, while being an environmentally friendly alternative. This article provides a brief overview of Antarctic bacterias, both psychrophilic and psychrotolerant that synthesis nanoparticles, possible mechanisms associated to this synthesis and future perspectives related to bacterial biosynthesis of nanoparticles.

Ceramides and terpenoids from Russula austrodelica Singer / Ceramidas y terpenoides desde Russula austrodelica Singer

A mixture of ceramides and known terpenes, was obtained from the fruiting bodies of Russula austrodelica. The structures were determined from chemical and spectroscopic evidence. R. austrodelica is a mycorrhizal fungus that grow in the Nothophagus forests of southern Chile. This is the first report of the isolation of ceramides in Chilean mushrooms.

Una mezcla de ceramidas y de terpenos conocidos, se obtuvo de los cuerpos fructíferos de Russula austrodelica. Las estructuras fueron determinadas a partir de evidencias químicas y espectroscópicas. R. austrodelica es un hongo micorrícico que crecen en los bosques de Nothophagus del sur de Chile. Este es el primer informe del aislamiento de ceramidas en hongos chilenos.

Insect growth regulatory activity of Blechnum chilense.

The genus Blechnum has 13 species that are common plants, well-distributed in Chile. Here, we report a phytochemical analysis of B. chilense (Kaulf.) Mett., as well as the insecticidal effects of extracts of this plant. From the n-hexane fraction four phytoecdysones were isolated: ecdysone, ponasterone, shidasterone and 2-deoxycrustecdysone. A bioassay with Drosophila melanogaster larvae was used to evaluate insecticidal activity. The EtOAc and n-hexane fractions at 800 ppm caused 66.7 and 50.0% larval mortality, respectively. Treatments with both extracts at 800 ppm caused the greatest larval mortality, whereas treatments with 500 and 200 ppm induced premature pupation compared with the control and the highest adult mortality, probably due to interference with ecdysteroid metabolism and inhibition of ecdysis triggering hormone (ETH). The dead adult flies exhibited malformations.

Solubility effects on antibacterial activity of chemically modified chitooligosaccharides of fungal origin / Efectos de la solubilidad sobre la actividad antibacteriana de quitooligosacáridos modificados químicamente de origen fúngico

Chitin and chitosan are a class of metabolites that occurring in some fungi species that are associated with commercial and medicinal plants, this is in Mucor sp. for example with an ample number of biological activities, being antibacterial and antifungal one of the most important. Into our program of search of biopesticides and natural compounds with biological activities, we have studying chitosan that was obtained from the culture medium of the fungus Mucor ruoxii. Chitooligosaccharides were prepared by partial acid hydrolysis of native chitosan and an aminoglycosylated derivative was obtained by reductive amination of the chitooligosaccaride. The solubilities of these compounds were measured at different pHs and its antibacterial activity against Escherichia coli (gram-negative) and Staphylococcus aureus (gram-positive). Chitosan and the derivatives tested exhibited a good antibacterial activity against S. aureus.

Quitina y quitosano son una clase de metabolitos que producen algunas especies de hongos que están asociados con plantas medicinales y comerciales, esto es por ejemplo en Mucor sp., con un amplio número de actividades biológicas, siendo la antibacteriana y antifúngica unas de las más importantes. En nuestro programa de investigación de biopesticidas y compuestos naturales, estamos estudiando quitosano obtenido de el medio de cultivo del hongo Mucor ruoxii. Quitooligosacáridos fueron preparados por hidrólisis parcial ácida de quitosano nativo y un derivado aminoglicosilado fue obtenido por aminación reductiva del quitooligosacárido. Las solubilidades de estos compuestos fueron medidas a diferentes pHs y su actividad antibacteriana frente a Escherichia coli (gram-negative) and Staphylococcus aureus (gram-positive). Quitosano y los derivados testeados exhiben una buena actividad antibacteriana frente a S. aureus.