RESUMO
The use of substances or conditions as elicitors can significantly increase the production of secondary metabolites. In this research, the effects of different elicitors on the production of antioxidant secondary metabolites were evaluated in a strain of Ganoderma sp. The elicitors tested were pH changes in different growth phases of the fungus (pH 3, 5.5 and 8), different concentrations of peptone as a nitrogen source (1 g/L and 10 g/L), and the addition of chemical agents to the culture medium (ethanol, growth regulators, and salts). The alkaline pH during the stationary phase and the high availability of nitrogen were effective elicitors, producing cultures with higher antioxidant activity (37.87 g/L and 43.13 g/L dry biomass) although there were no significant differences with other treatments.
Assuntos
Antioxidantes , Ganoderma , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Costa Rica , Ganoderma/metabolismo , NitrogênioRESUMO
The effects of ethanol/broth proportions and the number of steps at varying pH in the presence or absence of sodium chloride (NaCl) were studied as precipitation strategies for the recovery and purification of high molar mass bio-hyaluronic acid (Bio-HA). Bio-HA was synthesized by Streptococcus zooepidemicus in a culture medium containing glucose and soy peptones. A single-step precipitation was more attractive than multistep precipitation in terms of recovery and purity as well as decreased use of ethanol. The best conditions in the absence and presence of salt were 2:1 ethanol/broth (v/v) at pH 4 (55.0 ± 0.2% purity and 85.0 ± 0.7% recovery) and 2:1 ethanol/broth (v/v) at pH 7 + 2 mol L-1 NaCl (59.0 ± 0.9% purity and 82.0 ± 4.3% recovery). Dynamic light scattering (DLS) spectra showed different particle sizes as a consequence of the changes in the molecular structure of HA, mainly with changes in pH. Although slight changes in distribution were observed, the average HA molar mass was not affected by the precipitation strategy, remaining on the order of 105 Da. Therefore, pH and NaCl modulated the precipitation performance of HA. These findings are relevant to further optimizing the precipitation step, thus minimizing costs in the later stages of HA purification.
Assuntos
Ácido Hialurônico/isolamento & purificação , Biotecnologia , Precipitação Química , Meios de Cultura , Etanol , Fermentação , Ácido Hialurônico/biossíntese , Ácido Hialurônico/química , Concentração de Íons de Hidrogênio , Estrutura Molecular , Peso Molecular , Tamanho da Partícula , Cloreto de Sódio , Streptococcus equi/metabolismoRESUMO
This work addresses the production of prodigiosin from ram horn peptone (RHP) using MO-1, a local isolate in submerged culture. First, a novel gram-negative and rod-shaped bacterial strain, MO-1, was isolated from the body of the grasshopper (Poecilemon tauricola Ramme 1951), which was collected from pesticide-contaminated fields. Sequence analysis of 16S rDNA classified the microbe as Serratia marcescens. The substrate utilization potential (BIOLOG) and fatty acid methyl ester profile (FAME) of S. marcescens were also determined. The effect of RHP on the production of prodigiosin by S. marcescens MO-1 was investigated, and the results showed that RHP supplementation promoted the growth of MO-1 and increased the production of prodigiosin. A concentration of 0.4% (w/v) RHP resulted in the greatest yield of prodigiosin (277.74 mg/L) after 48 h when mannitol was used as the sole source of carbon. The pigment yield was also influenced by the types of carbon sources and peptones. As a result, RHP was demonstrated to be a suitable substrate for prodigiosin production. These results revealed that prodigiosin could be produced efficiently by S. marcescens using RHP.
Assuntos
Meios de Cultura/química , Peptonas/metabolismo , Prodigiosina/metabolismo , Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/metabolismo , Animais , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Gafanhotos/microbiologia , Dados de Sequência Molecular , Filogenia , Pigmentos Biológicos/metabolismo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Serratia marcescens/classificação , Serratia marcescens/isolamento & purificaçãoRESUMO
Streptococcus pneumoniae is a human pathogen largely transmitted by aerosols. Vaccines are the main strategy against this pathogen, and the capsular polysaccharide (PS) is its major antigen. S. pneumoniae serotype 1 is associated with large outbreaks and epidemics of invasive diseases. The aims of this work were to screen serotype 1 strains to identify the best PS1 producer, evaluate three peptones for PS1 production, investigate the effects of culture medium components using a design of experiments (DoE), a statistic tool for optimization, and propose a new medium/cultivation strategy. After flask cultivation of nine strains, two that produced high PS1 and biomass values were chosen for further evaluation in the bioreactor, and ST595/01 was chosen as the best PS1 producer strain. Among the peptones tested (Casamino acids, Soytone, and Phytone), the highest PS1 production (298 mg/L) was reached with Phytone. Next, DoE (2(4-1)) was performed to evaluate the effects of yeast extract (YE), Phytone, L-asparagine (Asn), and L-glutamine (Gln), yielding the following results: Phytone presented positive effects (p < 0.05) for maximum production of biomass, PS1, acetate, and lactate; YE showed positive effects for biomass and acid production (p < 0.05); Gln exerted a minor positive effect on PS1 yield factor on glucose (p < 0.1); and Asn presented only an effect on acetate production (p < 0.1). Hence, a new culture medium was formulated based on Phytone, YE, and glucose, and batch and fed-batch cultivations were evaluated. The fed-batch cultivation showed almost 2 times the biomass and 2.5 times the PS1 production as the batch culture, and 8-10 times higher PS1 production than has been previously reported.
Assuntos
Polissacarídeos Bacterianos/metabolismo , Streptococcus pneumoniae/crescimento & desenvolvimento , Streptococcus pneumoniae/metabolismo , Técnicas de Cultura Celular por Lotes , Meios de Cultura/química , Seleção Genética , SorogrupoRESUMO
This work addresses the production of prodigiosin from ram horn peptone (RHP) using MO-1, a local isolate in submerged culture. First, a novel gram-negative and rod-shaped bacterial strain, MO-1, was isolated from the body of the grasshopper (Poecilemon tauricola Ramme 1951), which was collected from pesticide-contaminated fields. Sequence analysis of 16S rDNA classified the microbe as Serratia marcescens. The substrate utilization potential (BIOLOG) and fatty acid methyl ester profile (FAME) of S. marcescens were also determined. The effect of RHP on the production of prodigiosin by S. marcescens MO-1 was investigated, and the results showed that RHP supplementation promoted the growth of MO-1 and increased the production of prodigiosin. A concentration of 0.4% (w/v) RHP resulted in the greatest yield of prodigiosin (277.74 mg/L) after 48 h when mannitol was used as the sole source of carbon. The pigment yield was also influenced by the types of carbon sources and peptones. As a result, RHP was demonstrated to be a suitable substrate for prodigiosin production. These results revealed that prodigiosin could be produced efficiently by S. marcescens using RHP.(AU)
Assuntos
Animais , Meios de Cultura/química , Peptonas/metabolismo , Prodigiosina/metabolismo , Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/metabolismo , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Gafanhotos/microbiologia , Dados de Sequência Molecular , Filogenia , Pigmentos Biológicos/metabolismo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Serratia marcescens/classificação , Serratia marcescens/isolamento & purificaçãoRESUMO
This work addresses the production of prodigiosin from ram horn peptone (RHP) using MO-1, a local isolate in submerged culture. First, a novel gram-negative and rod-shaped bacterial strain, MO-1, was isolated from the body of the grasshopper (Poecilemon tauricola Ramme 1951), which was collected from pesticide-contaminated fields. Sequence analysis of 16S rDNA classified the microbe as Serratia marcescens. The substrate utilization potential (BIOLOG) and fatty acid methyl ester profile (FAME) of S. marcescens were also determined. The effect of RHP on the production of prodigiosin by S. marcescens MO-1 was investigated, and the results showed that RHP supplementation promoted the growth of MO-1 and increased the production of prodigiosin. A concentration of 0.4% (w/v) RHP resulted in the greatest yield of prodigiosin (277.74 mg/L) after 48 h when mannitol was used as the sole source of carbon. The pigment yield was also influenced by the types of carbon sources and peptones. As a result, RHP was demonstrated to be a suitable substrate for prodigiosin production. These results revealed that prodigiosin could be produced efficiently by S. marcescens using RHP.
Assuntos
Animais , Meios de Cultura/química , Peptonas/metabolismo , Prodigiosina/metabolismo , Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/metabolismo , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Gafanhotos/microbiologia , Dados de Sequência Molecular , Filogenia , Pigmentos Biológicos/metabolismo , /genética , Análise de Sequência de DNA , Serratia marcescens/classificação , Serratia marcescens/isolamento & purificaçãoRESUMO
Heterotrophic CaCO3-precipitating bacteria were isolated from biofilms on deteriorated ignimbrites, siliceous acidic rocks, from Morelia Cathedral (Mexico) and identified as Enterobacter cancerogenus (22e), Bacillus sp. (32a) and Bacillus subtilis (52g). In solid medium, 22e and 32a precipitated calcite and vaterite while 52g produced calcite. Urease activity was detected in these isolates and CaCO3 precipitation increased in the presence of urea in the liquid medium. In the presence of calcium, EPS production decreased in 22e and 32a and increased in 52g. Under laboratory conditions, ignimbrite colonization by these isolates only occurred in the presence of calcium and no CaCO3 was precipitated. Calcium may therefore be important for biofilm formation on stones. The importance of the type of stone, here a siliceous stone, on biological colonization is emphasized. This calcium effect has not been reported on calcareous materials. The importance of the effect of calcium on EPS production and biofilm formation is discussed in relation to other applications of CaCO3 precipitation by bacteria.
Assuntos
Bacillus/fisiologia , Biofilmes , Carbonato de Cálcio/metabolismo , Enterobacter/fisiologia , Bacillus/genética , Bacillus/isolamento & purificação , Carbonato de Cálcio/química , Precipitação Química , Enterobacter/genética , Enterobacter/isolamento & purificação , Processos Heterotróficos , México , Dados de Sequência Molecular , Filogenia , Propriedades de SuperfícieRESUMO
Foram avaliados 25 reservatórios de água dos equipos odontológicos da Clínica de Dentística/Endodontia da FOB/USP com relação à presença de micro-organismos e a ação do ultrassom (US) na remoção do biofilme. Amostras de 10ml de água foram obtidas e alíquotas de 25l in natura e diluída até 10-4 foram semeadas pela técnica da gota nos meios: R2A Agar (R2A), Plate Count Agar (PCA), Peptona Diluída (PD) e Sabouraud Dextrose Agar com cloranfenicol a 1% (SDA), incubadas a 24º C por 72 horas. A água dos reservatórios foi descartada e 500 ml de água destilada esterilizada foi adicionada, sendo submetidos à ação do ultrassom (US) por 15 minutos, seguidos do mesmo procedimento descrito anteriormente. As colônias de bactérias foram quantificadas e os fungos foram identificados por micro-cultivo. A média da detecção de UFC/ml antes e após o US foi de 173.787 e 15.841 para o R2A, 104.873 e 3.034 para o PCA e de 245.824 e 8.231 para o PD. A média de fungos foi de 52,4 antes e 19,2 UFC/ml após ação do US. Fungos foram detectados em 20 reservatórios antes e em 12 deles após uso do US. O Penicillium sp apresentou prevalência de 36% nos reservatórios de água avaliados. Os resultados obtidos permitem concluir que o US foi eficiente em desestruturar o biofilme, embora não o elimine por completo, apresentando maior efetividade na desestruturação de bactérias.
A total of 25 waterline unit reservoirs of the odontological sets from the Dentistry/Endodontic Clinic of FOB/USP were assessed, in relation to the presence of microorganisms and the ultrasound action (US) on the biofilm removal. Waterline samples of 10ml were obtained from aliquots of 25l in natura and diluted until 10-4, then, they were spread using the dripping technique on the means: R2A Agar (R2A), Plate Count Agar (PCA), diluted Peptone (DP) and Sabouraud Dextrose Agar with cloranfenicol at 1% (SDA), being incubated at 24º C for 72 hours. The waterline units of the reservoirs were discarded and 500 ml of sterilized distilled water was added, submitted to ultrasound action (US) for 15 minutes, following the same procedure described afore. The bacteria colonies were quantified and the fungi were identified through micro-culture. The average of detection of UFC/ml before and after US was 173.787 and 15.841 for R2A, 104.873 and 3.034 for PCA and of 245.824 and 8.231 for PD. The fungi average was 52,4 before and 19,2 UFC/ml after the action of US. Fungi were detected in 20 reservoirs before and 12 after using US. Penicillium sp showed a prevalence of 36% in the waterline reservoirs assessed. The results obtained, led to the conclusion that US was efficient to break the structure of the biofilm, although it did not eliminate it completely, showing more effectiveness to break the bacteria structure.