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1.
Braz. j. biol ; 83: e250550, 2023. tab, graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1345536

ABSTRACT

Abstract Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.


Resumo A vanilina é o principal componente responsável pelo sabor e aroma do extrato de baunilha e é produzida de três formas: extração natural da planta da baunilha, síntese química e transformação microbiana. A pesquisa atual teve como objetivo estudar a produção bacteriana de vanilina a partir de fontes naturais nativas, incluindo esgoto e solo de áreas industriais. O objetivo principal era a bioprodução de vanilina por meio do isolamento de bactérias dessas fontes nativas. Também para adaptar metodologias para melhorar a produção de vanilina por meio de fermentação otimizada e condições de crescimento. Foram coletadas 47 amostras de solo e 13 de esgoto de diferentes regiões industriais de Lahore, Gujranwala, Faisalabad e Kasur; 67,7% dos isolados bacterianos produziram vanilina e 32,3% eram não produtores. Desses 279 produtores, 4 isolados bacterianos selecionados como produtores significativos foram: A3, A4, A7 e A10. Esses isolados foram identificados por ribotipagem como fluorescência A3 Pseudomonas (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) e A10 Bacillus subtilis (KT962919). Os produtores de vanilina foram posteriormente testados para produção aprimorada de vanilina e foram cultivados em diferentes meios de fermentação sob condições de crescimento otimizadas para produção aprimorada de vanilina. Os meios de fermentação (FM) foram: à base de óleo de cravo, à base de resíduos de farelo de arroz (resíduos de óleo), à base de farelo de trigo e à base de isoeugenol modificado. Em FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36 e FM37, as 4 cepas bacterianas selecionadas produziram quantidades significativas de vanilina. A10 B. subtilis produziu quantidade máxima de vanilina. Essa cepa produziu 17,3 g / L de vanilina em FM36. O custo desse meio de fermentação 36 foi de 131,5 rúpias / L. Esse meio de fermentação foi um meio à base de isoeugenol modificado com 1% de isoeugenol e 2,5 g / L de farelo de soja. O gene ech foi amplificado em A3 P. fluorescence usando primers específicos para ech. Como o uso da vanilina como sabor aumentou tremendamente, a bioprodução da vanilina deve ser focada.


Subject(s)
Benzaldehydes/metabolism , Flavoring Agents/metabolism , Bacillus subtilis/metabolism , Industrial Microbiology , Pseudomonas fluorescens/metabolism , Enterococcus faecium/metabolism , Culture Media , Alcaligenes faecalis/metabolism , Fermentation
2.
Chinese Journal of Biotechnology ; (12): 4303-4313, 2021.
Article in Chinese | WPRIM | ID: wpr-921507

ABSTRACT

D-allulose-3-epimerase (DPEase) is the key enzyme for isomerization of D-fructose to D-allulose. In order to improve its thermal stability, short amphiphilic peptides (SAP) were fused to the N-terminal of DPEase. SDS-PAGE analysis showed that the heterologously expressed DPEase folded correctly in Bacillus subtilis, and the protein size was 33 kDa. After incubation at 40 °C for 48 h, the residual enzyme activity of SAP1-DSDPEase was 58%. To make the recombinant B. subtilis strain reusable, cells were immobilized with a composite carrier of sodium alginate (SA) and titanium dioxide (TiO2). The results showed that 2% SA, 2% CaCl2, 0.03% glutaraldehyde solution and a ratio of TiO2 to SA of 1:4 were optimal for immobilization. Under these conditions, up to 82% of the activity of immobilized cells could be retained. Compared with free cells, the optimal reaction temperature of immobilized cells remained unchanged at 80 °C but the thermal stability improved. After 10 consecutive cycles, the mechanical strength remained unchanged, while 58% of the enzyme activity could be retained, with a conversion rate of 28.8% achieved. This study demonstrated a simple approach for using SAPs to improve the thermal stability of recombinant enzymes. Moreover, addition of TiO2 into SA during immobilization was demonstrated to increase the mechanical strength and reduce cell leakage.


Subject(s)
Bacillus subtilis/metabolism , Carbohydrate Epimerases/genetics , Enzyme Stability , Enzymes, Immobilized/metabolism , Fructose , Hydrogen-Ion Concentration , Racemases and Epimerases , Temperature
3.
Chinese Journal of Biotechnology ; (12): 2377-2386, 2020.
Article in Chinese | WPRIM | ID: wpr-878494

ABSTRACT

Surfactin has great potential applications in enhancing oil recovery, agriculture, pharmaceuticals, foods and beverages, and cosmetics due to its extraordinary surface activity, biodegradability, anti-bacterial activity and biocompatibility. Enhancing surfactin production by engineering surfactin-producer and optimizing culture conditions is the key of its industrial production and subsequent applications. In this study, the effect of fatty acid synthesis pathway on surfactin synthesis was investigated, and Bacillus subtilis THBS-2 and THBS-8 with high surfactin titer were constructed by overexpressing key genes involved in the fatty acid synthesis pathway. To optimize culture condition, the amount and adding time of isopropyl-beta-D-thiogalactopyranoside (IPTG) and amino acids were studied, and a two-stage culture method was obtained: IPTG (final concentration: 1.25 mmol/L) and leucine (final concentration: 5 g/L) were added at 3 h, leucine (final concentration 5 g/L) and condensed culture medium (5 mL) were added at 24 h. Applying this strategy, the surfactin titer of B. subtilis THBS-2 reached to 24 g/L in shake flask at 48 h and up to 34 g/L after 68 h fermentation in a 30-L fermentor. The results provide basis for large-scale production and broad application of surfactin.


Subject(s)
Amino Acids , Bacillus subtilis/metabolism , Culture Media , Fermentation , Lipopeptides , Peptides, Cyclic
4.
Braz. j. microbiol ; 48(4): 809-814, Oct.-Dec. 2017. graf
Article in English | LILACS | ID: biblio-889176

ABSTRACT

ABSTRACT This study aimed to describe a Bacillus subtilis expression system based on genetically modified B. subtilis. Abaecin, an antimicrobial peptide obtained from Apis mellifera, can enhance the effect of pore-forming peptides from other species on the inhibition of bacterial growth. For the exogenous expression, the abaecin gene was fused with a tobacco etch virus protease cleavage site, a promoter Pglv, and a mature beta-glucanase signal peptide. Also, a B. subtilis expression system was constructed. The recombinant abaecin gene was expressed and purified as a recombinant protein in the culture supernatant. The purified abaecin did not inhibit the growth of Escherichia coli strain K88. Cecropin A and hymenoptaecin exhibited potent bactericidal activities at concentrations of 1 and 1.5 µM. Combinatorial assays revealed that cecropin A and hymenoptaecin had sublethal concentrations of 0.3 and 0.5 µM. This potentiating functional interaction represents a promising therapeutic strategy. It provides an opportunity to address the rising threat of multidrug-resistant pathogens that are recalcitrant to conventional antibiotics.


Subject(s)
Antimicrobial Cationic Peptides/genetics , Antimicrobial Cationic Peptides/metabolism , Bacillus subtilis/genetics , Genetic Vectors/genetics , Insect Proteins/genetics , Insect Proteins/metabolism , Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/pharmacology , Antimicrobial Cationic Peptides/isolation & purification , Antimicrobial Cationic Peptides/pharmacology , Bacillus subtilis/metabolism , Escherichia coli/drug effects , Escherichia coli/growth & development , Gene Expression , Genetic Vectors/metabolism , Insect Proteins/isolation & purification , Insect Proteins/pharmacology , Protein Engineering , Protein Transport , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Recombinant Proteins/pharmacology
5.
Electron. j. biotechnol ; 30: 33-38, nov. 2017. tab, graf
Article in English | LILACS | ID: biblio-1021336

ABSTRACT

Background: Lipases are used in detergent industries to minimise the use of phosphate-based chemicals in detergent formulations. The use of lipase in household laundry reduces environmental pollution and enhances the ability of detergent to remove tough oil or grease stains. Results: A lipase-producing indigenous Bacillus subtilis strain [accession no. KT985358] was isolated from the foothills of Trikuta mountain in Jammu and Kashmir, India. The lipase (BSK-L) produced by this strain expressed alkali and thermotolerance. Lipase has an optimal activity at pH 8.0 and temperature 37°C, whereas it is stable at pH 6.0­9.0 and showed active lipolytic activity at temperatures 30 to 60°C. Furthermore, lipase activity was found to be stimulated in the presence of the metal ions Mn2+, K+, Zn2+, Fe2+ and Ca2+. This lipase was resistant to surfactants, oxidising agents and commercial detergents, suggesting it as a potential candidate for detergent formulation. BSK-L displayed noticeable capability to remove oil stains when used in different washing solutions containing buffer, lipase and commercial detergent. The maximum olive oil removal percentage obtained was 68% when the optimum detergent concentration (Fena) was 0.3%. The oil removal percentage from olive oil-soiled cotton fabric increased with 40 U/mL of lipase. Conclusions: This BSK-L enzyme has the potential for removing oil stains by developing a pre-soaked solution for detergent formulation and was compatible with surfactants, oxidising agents and commercial detergents.


Subject(s)
Bacillus subtilis/metabolism , Lipase/metabolism , Temperature , Bacillus subtilis/isolation & purification , Bacillus subtilis/enzymology , Detergents , Alkalinization , Thermotolerance , Hydrogen-Ion Concentration , Lipase/biosynthesis
6.
Electron. j. biotechnol ; 27: 49-54, May. 2017. graf, tab
Article in English | LILACS | ID: biblio-1010292

ABSTRACT

Background: Surfactants are one of the most important raw materials used in various industrial fields as emulsifiers, corrosion inhibitors, foaming agents, detergent products, and so on. However, commercial surfactant production is costly, and its demand is steadily increasing. This study aimed to evaluate the performance of typical strains of Bacillus sp. to produce biosurfactants through fermentation. It also included the investigation of the effect of initial glucose concentration and the carbon to nitrogen ratio. Results: The biosurfactant yield was in the range of 1­2.46 g/L at initial glucose concentrations of 10­70 g/L. The optimum fermentation condition was achieved at a carbon to nitrogen ratio of 12.4, with a decrease in surface tension of up to 27 mN/m. Conclusions: For further development and industrial applications, the modified Gompertz equation is proposed to predict the cell mass and biosurfactant production as a goodness of fit was obtained with this model. The modified Gompertz equation was also extended to enable the excellent prediction of the surface tension.


Subject(s)
Surface-Active Agents/metabolism , Bacillus subtilis/metabolism , Surface-Active Agents/chemistry , Surface Tension , Bacillus subtilis/physiology , Carbon/analysis , Kinetics , Fermentation , Glucose/analysis , Micelles , Nitrogen/analysis
7.
São Paulo; s.n; s.n; 2017. 106 p. tab, ilus, graf.
Thesis in Portuguese | LILACS | ID: biblio-883613

ABSTRACT

Seja no meio ambiente, dentro de um hospedeiro ou em outro habitat, bactérias estarão frequentemente enfrentando condições adversas, como exposição a compostos antibacterianos ou carência nutricional. Em situações como essas, as bactérias são capazes de ativar a chamada resposta estringente, modulada pelo alarmônio (p)ppGpp. O acúmulo de (p)ppGpp promove a inibição da transcrição de rRNAs e tRNAs e a supressão do processo de tradução, e a ativação de operons de biossíntese de aminoácidos. Sabe-se também hoje que a resposta estringente está relacionada a outras importantes carências nutricionais em Escherichia coli, como a falta de ácidos graxos, porém não se sabe se o mesmo ocorre em Bacillus subtilis ou em outras Grampositivas. (p)ppGpp atua também direta e indiretamente em vários outros processos celulares, como motilidade, resistência a antibióticos, virulência e persistência, indicando que (p)ppGpp é um regulador central que integra informação metabólica e respostas adaptativas. O presente trabalho buscou estudar a correlação da resposta estringente de B. subtilis com a carência de ácidos graxos e a busca por pequenas moléculas capazes de modular RelA (a principal proteína envolvida na síntese de (p)ppGpp) e impedir o acúmulo de (p)ppGpp. Para a indução da carência de ácidos graxos, foram utilizadas duas estratégias; uso da droga Cerulenina (inibidor de FabF) e mutantes condicionais no gene FabF. Observou-se que mutantes incapazes de ativar a resposta estringente (cepa ppGpp(0) ou RelAD264G) apresentaram grande perda de viabilidade celular durante a carência de ácidos graxos, ao passo que a cepa selvagem manteve sua viabilidade celular. A causa da morte se deu majoritariamente devido ao colapso do potencial de membrana. Apesar de não termos observado aumento de (p)ppGpp nas células selvagens durante a carência de ácidos graxos, observou-se uma redução da razão GTP/ATP, ao passo que na cepa ppGpp(0), a razão GTP/ATP aumentou, devido ao acúmulo de GTP. O uso da droga decoinina, capaz de reduzir os níveis intracelulares de GTP, resgatou parcialmente a viabilidade da cepa e impediu a perda do potencial de membrana, indicando que os níveis de GTP são importantes durante a carência de ácidos graxos em B. subtilis. Para a triagem de pequenas moléculas inibidoras do acúmulo de (p)ppGpp, foi utilizada uma biblioteca de 2320 diferentes compostos químicos, e buscou-se drogas capazes de reverter o fenótipo de crescimento lento de cepas de B. subtilis que acumulam (p)ppGpp (via mutação pontual; mutante RelAH77A e via tratamento com o indutor hidroxamato de arginina) em meio rico. A primeira etapa selecionou 40 moléculas capazes de resgatar o crescimento de células tratadas com arginina-hidroxamato, porém apenas uma, salicilanilida, foi capaz de também resgatar o crescimento da cepa RelAH77A. Todavia, apesar de ser capaz de acelerar o crescimento de B. subtilis esse efeito é limitado. Diversos análogos de salicilanilida foram testados, porém não apresentaram efeito superior a salicilanilida para a reversão do fenótipo de crescimento lento de B. subtilis. Em adição, a droga não foi capaz de aumentar a sensibilidade dos organismos a diversos antibióticos testados, e aparentemente é incapaz de alterar os níveis internos de (p)ppGpp, porém é capaz de causar alterações nos níveis de ATP. Logo, acredita-se que o efeito observado para o crescimento das células seja devido a efeitos indiretos, possivelmente envolvendo alteração de outros nucleotídeos fosforilados


In the environment, inside a host or other habitat, bacteria will always face adverse conditions, as for example exposure to antimicrobials or starvation. In situations like those, bacteria activate the stringent response, modulated by the alarmone (p)ppGpp. (p)ppGpp accumulation promotes inhibition of rRNA and tRNA transcription and suppression of translational process, at the same time that it activates several amino acid biosynthesis operons. It is known also that the stringent response it is related to other starvation stress in Escherichia coli, like lack of fatty acids, but there is no knowledge if the same occurs for Bacillus subtilis or other gram-positive bacteria. ppGpp acts directly and indirectly affecting several other cellular process, as motility, resistance to antibiotics, virulence and persistence, indicating that (p)ppGpp is a central regulator that integrates metabolic information and adaptive responses. This work aimed to study the correlation between the stringent response in B. subtilis with fatty acid starvation, and search for small moleculas capable of modulating RelA (the main enzyme responsible for ppGpp synthesis) and stop (p)ppGpp production. For fatty acid starvation induction, two strategies were used; use of the drug Cerulenin (inhibitor of the FabF protein) and conditional mutants of the FabF gene. We observed that mutants incapable of activating the stringent response (strains ppGpp(0) ou RelAD264G) presented great loss of viability during fatty acid starvation, whereas the wild-type strain keeps its viability. The main cause of death is due membrane rupture in some cells, but mainly due to membrane potential collapse. Although we did not observed increase of (p)ppGpp in wild-type strains during fatty acid starvation, we observed reduction in GTP/ATP ratios, a hallmark of (p)ppGpp production in gram-positive bacteria. In the strain ppGpp(0) GTP/ATP ratio increased, mainly due to GTP increase. Using the drug decoyinine, capable of reducing GTP levels, partially recued viability and protects cells of losing its membrane potential, indicating that GTP levels plays an important role during fatty acid starvation in B. subtilis. For the screening of small molecules capable of inhibit (p)ppGpp production, a library of 2320 different chemical compounds were used, and we looked for drugs capable of reverting the slow growth phenotype of B. subtilis strains with (p)ppGpp accumulation (using a mutant RelAH77A; and using a stringent response inductor, arginine hidroxamate). The first step selected for 40 molecules capable of rescuing the growth of cells treated with arginine hidroxamate, but only one drug, salicilanilyde could also rescue the growth of the strain RelAH77A. Although capable of rescuing growth of B. subtilis that accumulates (p)ppGpp, this rescue is limited. Several analogues of salicilanilyde were tested, but none were stronger than salicilanilyde itself in rescuing growth of slow growing strains of B. subtilis. In addition, the drug was not capable of increasing antibiotic sensibility and it is incapable of changing intracellular (p)ppGpp levels, but it does shifts ATP levels. Therefore, we believe that the observed effects of salicilanilyde is due indirect action, probably involving other phosphorylated nucleotides, rather than modifying (p)ppGpp levels


Subject(s)
Bacillus subtilis/metabolism , Transcription Factor RelA , Cerulenin/administration & dosage , Chromatography, High Pressure Liquid , Fatty Acids/analysis , Microbial Sensitivity Tests/methods , Microscopy, Fluorescence/instrumentation , Salicylanilides/administration & dosage , Triage/statistics & numerical data
8.
Braz. j. microbiol ; 47(4): 955-964, Oct.-Dec. 2016. tab, graf
Article in English | LILACS | ID: biblio-828202

ABSTRACT

Abstract The possible application of a bacterial strain - Bacillus subtilis R1, isolated from an oil contaminated desert site in India, as biocontrol agent and its biosurfactant in microbial enhanced oil recovery are discussed. The biosurfactant production in minimal medium was carried out at different temperatures and salt concentrations, where it produced an efficient biosurfactant at 30-45 °C and in presence of up to 7% salt. It significantly reduced the surface tension from 66 ± 1.25 mN/m to 29 ± 0.85 mN/m within 24 h. In order to enhance the biosurfactant production, random mutagenesis of B. subtilis R1 was performed using chemical mutagen - ethyl methanesulfonate. Majority of the isolated 42 mutants showed biosurfactant production, but the difference was statistically insignificant as compared with parent strain R1. Therefore none of the mutants were selected for further study, and only parent strain R1 was studied. The biosurfactant was quite stable under harsh conditions for up to 10 days. The biosurfactant was extracted and characterized as similar to the lipopeptide group - surfactins and fengycin. The crude oil displacement experiments using biosurfactant broth in sand pack glass columns showed 33 ± 1.25% additional oil recovery. The strain also showed inhibition of various plant pathogenic fungi on potato dextrose agar medium.


Subject(s)
Bacillus subtilis/metabolism , Lipopeptides/biosynthesis , Surface-Active Agents/metabolism , Surface-Active Agents/pharmacology , Bacillus subtilis/classification , Bacillus subtilis/genetics , RNA, Ribosomal, 16S/genetics , Microbial Sensitivity Tests , Mutagenesis , Spectroscopy, Fourier Transform Infrared , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Lipopeptides/pharmacology , Metabolic Engineering , Hydrogen-Ion Concentration , Antifungal Agents/metabolism , Antifungal Agents/pharmacology
9.
Braz. j. microbiol ; 47(3): 665-674, July-Sept. 2016. tab, graf
Article in English | LILACS | ID: lil-788963

ABSTRACT

ABSTRACT The simultaneous production of amylase (AA) and protease (PA) activity by Bacillus subtilis UO-01 in brewery wastes was studied by combining the response surface methodology with the kinetic study of the process. The optimum conditions (T = 36.0 °C and pH = 6.8) for high biomass production (0.92 g/L) were similar to the conditions (T = 36.8 °C and pH = 6.6) for high AA synthesis (9.26 EU/mL). However, the maximum PA level (9.77 EU/mL) was obtained at pH 7.1 and 37.8 °C. Under these conditions, a considerably high reduction (between 69.9 and 77.8%) of the initial chemical oxygen demand of the waste was achieved. In verification experiments under the optimized conditions for production of each enzyme, the AA and PA obtained after 15 h of incubation were, respectively, 9.35 and 9.87 EU/mL. By using the Luedeking and Piret model, both enzymes were classified as growth-associated metabolites. Protease production delay seemed to be related to the consumption of non-protein and protein nitrogen. These results indicate that the brewery waste could be successfully used for a high scale production of amylases and proteases at a low cost.


Subject(s)
Peptide Hydrolases/biosynthesis , Bacillus subtilis/metabolism , Fermentation , Amylases/biosynthesis , Industrial Waste , Temperature , Kinetics , Biotransformation , Carbohydrate Metabolism , Hydrogen-Ion Concentration
10.
Indian J Exp Biol ; 2014 Jul; 52(7): 692-704
Article in English | IMSEAR | ID: sea-153749

ABSTRACT

The physiological role of C-reactive protein (CRP), the classical acute-phase protein, is not well documented, despite many reports on biological effects of CRP in vitro and in model systems in vivo. It has been suggested that CRP protects mice against lethal toxicity of bacterial infections by implementing immunological responses. In Achatina fulica CRP is a constitutive multifunctional protein in haemolymph and considered responsible for their survival in the environment for millions of years. The efficacy of Achatina CRP (ACRP) was tested against both Salmonella typhimurium and Bacillus subtilis infections in mice where endogenous CRP level is negligible even after inflammatory stimulus. Further, growth curves of the bacteria revealed that ACRP (50 µg/mL) is bacteriostatic against gram negative salmonellae and bactericidal against gram positive bacilli. ACRP induced energy crises in bacterial cells, inhibited key carbohydrate metabolic enzymes such as phosphofructokinase in glycolysis, isocitrate dehydrogenase in TCA cycle, isocitrate lyase in glyoxylate cycle and fructose-1,6-bisphosphatase in gluconeogenesis. ACRP disturbed the homeostasis of cellular redox potential as well as reduced glutathione status, which is accompanied by an enhanced rate of lipid peroxidation. Annexin V-Cy3/CFDA dual staining clearly showed ACRP induced apoptosis-like death in bacterial cell population. Moreover, immunoblot analyses also indicated apoptosis-like death in ACRP treated bacterial cells, where activation of poly (ADP-ribose) polymerase-1 (PARP) and caspase-3 was noteworthy. It is concluded that metabolic impairment by ACRP in bacterial cells is primarily due to generation of reactive oxygen species and ACRP induced anti-bacterial effect is mediated by metabolic impairment leading to apoptosis-like death in bacterial cells.


Subject(s)
Animals , Anti-Bacterial Agents/pharmacology , Apoptosis/drug effects , Bacillus subtilis/drug effects , Bacillus subtilis/metabolism , C-Reactive Protein/isolation & purification , C-Reactive Protein/pharmacology , Gluconeogenesis/drug effects , Glycolysis/drug effects , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/metabolism , Gram-Negative Bacterial Infections/microbiology , Hemolymph/metabolism , Homeostasis/drug effects , Immunoblotting , Lipid Peroxidation/drug effects , Male , Mice , Oxidation-Reduction , Reactive Oxygen Species/metabolism , Salmonella Infections/drug therapy , Salmonella Infections/metabolism , Salmonella Infections/microbiology , Salmonella typhimurium/drug effects , Salmonella typhimurium/metabolism , Snails
11.
International Journal of Environmental Research. 2013; 7 (4): 851-858
in English | IMEMR | ID: emr-138318

ABSTRACT

In recent years, biosurfactants due to wide applications in chemical, petroleum, food and pharmaceutical industries, have been widely considered by researchers. Biosurfactants are produced by a series of microorganisms, so it is important to screen culture medium and operating conditions in miniaturized bioreactors prior to scaling up to large bioreactors.In this study, using a kind of miniaturized bioreactor called ventilation flask, optimal production conditions, including filling volume and shaking frequency to produce a surfactin-type biosurfactant by Bacillus subtilis ATCC 6633, were examined. Moreover, the effect of oxygen transfer rate [OTR] on the surfactin production was investigated according to Amoabediny and Buchs model. The results indicated that the maximum biomass and biosurfactant yield which obtained under optimal conditions [filling volume of 15 mL and shaking frequency of 300 rpm] were evaluated 0.3 g/L/h and 0.0485 g/L/h, respectively. Also, at the same conditions, the amount of surface tension decreased from 60.5 mN/m to 31.7 mN/m and the maximum oxygen transfer rate [OTR[max]] obtained as 0.01 mol/L/h


Subject(s)
Lipopeptides/biosynthesis , Polysaccharides, Bacterial/biosynthesis , Bacillus subtilis/metabolism , Bioreactors
12.
Article in English | IMSEAR | ID: sea-136347

ABSTRACT

Background & objectives: A cyclic lipopeptide, surfactin produced by a strain of Bacillus subtilis subsp. subtilis (VCRC B471) was found to exhibit activity against both the larval and pupal stages of mosquitoes. The present study was aimed at increasing the production of the mosquitocidal metabolite by modifying the conventional medium. Methods: Enhancement of mosquitocidal metabolite production was attempted by replacing the existing micronutrients of the conventional NYSM and supplementing the medium with additional amounts of glucose. The LC50 value of culture supernatant (CS) against the larval and pupal stages of Anopheles stephensi was determined. Crude mosquitocidal metabolite (CMM) was separated from the CS, identified by MALDI-TOF analysis and its LC50 dosage requirement for the pupal stage of the above mosquito species determined. Results: The medium containing a new composition of micronutrients and glucose up to 1 per cent resulted in increased metabolite production. The LC50 value of the CS obtained in the improved medium against larvae and pupae of An. stephensi was 5.57 and 0.71 μl/ml, respectively. The yield of CMM was doubled in the improved medium. MALDI-TOF analysis revealed that the CMM was surfactin. Interpretation & conclusions: The new improved medium enhanced the production of mosquitocidal metabolite as the dosage required for inciting 50 per cent mortality among the pupal stages of mosquitoes was only half of that required when the metabolite was produced in the conventional medium. The mosquitocidal metabolite was identified as surfactin, a cyclic lipopeptide and biosurfactant.


Subject(s)
Animals , Bacillus subtilis/growth & development , Bacillus subtilis/metabolism , Bacterial Proteins/biosynthesis , Bacterial Proteins/chemistry , Bacterial Proteins/pharmacology , Culicidae/drug effects , Culture Media/chemistry , Humans , Insecticides , Lipopeptides/biosynthesis , Lipopeptides/chemistry , Lipopeptides/pharmacology , Peptides, Cyclic/biosynthesis , Peptides, Cyclic/chemistry , Peptides, Cyclic/pharmacology
13.
Electron. j. biotechnol ; 13(4): 6-7, July 2010. ilus, tab
Article in English | LILACS | ID: lil-577111

ABSTRACT

Response surface methodology (RSM) was used for statistical optimization of jiean-peptide (JAA) production by Bacillus subtilis ZK8 cells adsorbed on wood chips to form a novel fermentation system. The Plackett-Burman design was used in the first step to evaluate the effects of eight factors, including six fermentation medium components and two cell adsorption conditions. Among the variables screened, soybean meal hydrolysate (SMH) and MgSO4A7H2O in the fermentation medium had significant effects on JAA production. In the second step, the concentrations of SMH and MgSO4A7H2O were further optimized using central composite designs and response surface analysis. The optimized concentration of SMH and MgSO4À7H2O was 24 percent (v/v) and 0.38 percent (w/v), respectively, which increased the production of JAA in a shake flask system by 41 percent relative to optimization of a single variable component of the culture medium.


Subject(s)
Bacillus subtilis/metabolism , Fermentation , Wood/metabolism , Peptides/metabolism , Analysis of Variance , Culture Media , Models, Biological
14.
Indian J Exp Biol ; 2009 Jan; 47(1): 57-62
Article in English | IMSEAR | ID: sea-56827

ABSTRACT

Among different bacterial cultures, a potent Bacillus subtilis MTCC-8114 was isolated from garden soil samples which showed 16 and 14 mm inhibition zones by spot inoculation method and 24 and 22 mm inhibition zones by well agar diffusion method against test fungi i.e. Microsporum fulvum and Trichophyton species. Among four media tested, the maximum growth and antibiotic production was found in trypticase soya broth (TSB) medium at 37 degrees C, pH-7 and 48 h of incubation. The Rf value (0.64) by Thin Layer Chromatography (TLC) technique and UV and FTIR spectral data of the active antifungal compound, indicated that the isolated compound belongs to peptide antifungal antibiotic group. MIC value of antifungal antibiotic was 135 and 145 microg/ml.


Subject(s)
Antifungal Agents/biosynthesis , Antifungal Agents/pharmacology , Bacillus subtilis/drug effects , Bacillus subtilis/growth & development , Bacillus subtilis/metabolism , Culture Media , Microbial Sensitivity Tests , Microsporum/drug effects , Peptide Biosynthesis/drug effects , Pest Control, Biological , Spectrophotometry, Ultraviolet , Spectroscopy, Fourier Transform Infrared , Trichophyton/drug effects
15.
Article in English | WPRIM | ID: wpr-96781

ABSTRACT

Platelet aggregation was inhibited and the density of platelet-rich plasma (PRP) clots was decreased by the preincubation of PRP with surfactins, an acidic lipopeptide of Bacillus subtilis complex BC1212 isolated from soybean paste, in dose-dependent manner. Our findings suggest that surfactins are able to prevent a platelet aggregation leading to an inhibition of additional fibrin clot formation, and to enhance fibrinolysis with facilitated diffusion of fibrinolytic agents.


Subject(s)
Bacillus subtilis/metabolism , Blood Platelets/drug effects , Fibrinolytic Agents/pharmacology , Humans , Peptides, Cyclic , Platelet Aggregation Inhibitors/pharmacology
16.
Rev. microbiol ; 29(4): 259-64, out.-dez. 1998. tab, graf
Article in English | LILACS | ID: lil-251733

ABSTRACT

The influence of aeration and automatic pH control on the production of alpha-amylase by a strain of "Bacillus subtilis" NRRL 3411 from acid cheese whey was studied. Tests were carried out in a rotary shaker and in mechanically stirred ferments. Alpha-maylase was analysed according to DUN's method. Oxygen absorption rate was determined by Cooper's method. Cell oxygen demand was determined as oxygen consumption in a Warburg respirometer. The level of dissolved oxygen was mesured by means of a galvanic silver-lead electrode. Results suggest the possibility of industrial use of acid cheese whey as a carbon source for alpha-amylase production, since the yiels was similar to that produced with lactose. The highest alpha-amylase levels 100,000 DUN/ml units were not attained at highr aeration rates -431 mLO2/L.h-. The indicated value correspond to a 96 h process with automatic pH enzyme production was directly related to growth in the form of cell aggregates.


Subject(s)
alpha-Amylases/biosynthesis , Bacillus subtilis/metabolism , Cheese/microbiology , Bacillus subtilis/enzymology , Culture Media, Serum-Free/analysis , Oxygen Consumption
17.
Indian J Exp Biol ; 1998 Jun; 36(6): 631-4
Article in English | IMSEAR | ID: sea-63195

ABSTRACT

Phytoalexins from four different treatments viz. control, AF 1-treated, A. niger-treated, and dual inoculated were separated by TLC showed that one phytoalexin with an Rf value of 0.628 (P1) appeared on 2nd day only in dual-inoculated seeds of groundnut (A. hypogaea). By 3rd day three additional phytoalexins were visualized in response to A. niger-treatment with lower Rf values 0.485 (P2), 0.388 (P3) and 0.314 (P4) as compared to P1. In dual inoculated seedlings, P1 and P3 could be visualized while only P1 appeared in response to AF 1 on 3rd day. All the compounds lost fluorescence on exposure to light, got converted to pale yellow colour. In all the treatments no phytoalexin accumulation was observed after 3rd day. All the four phytoalexins had a major peak between 338 and 339.5 nm. In potato dextrose broth, the growth of A. niger showed a steady increase up to 32 hr while it was significantly inhibited with P1 in microtiter plates. P2, P3 and P4 (in the same order) had significantly less antifungal activity as compared to P1. The antifungal activity of the phytoalexins decreased with decrease in their Rf value.


Subject(s)
Aspergillus niger/metabolism , Bacillus subtilis/metabolism , Nuts/microbiology , Plant Diseases/microbiology , Plant Extracts/metabolism , Terpenes
18.
Rev. microbiol ; 26(1): 46-51, jan.-mar. 1995. tab, graf
Article in Portuguese | LILACS | ID: lil-283828

ABSTRACT

Foi investigado o efeito da concentraçäo inicial de diferentes fontes de carbonos sobre a produçäo de amilase por uma linhagem de Bacillus subtilis. O microorganismo produziu alfa-amilase em meios de cultura contendo glicose, amido solúvel, dextrina e lactose. Foi observado que quanto maiores as concentraçöes iniciais de carboidrato maiores foram os tempos de fermentaçäo necessários para ocorrer a máxima velocidade específica de produçäo da enzima. As maiores produçöes de alfa-amilase ocorreram em meios de cultura contendo glicose a 0.75 g/L e 30.0 g/L ou dextrina a 10.0 g/L. Resultados obtidos para a atividade enzimática específica global mostraram que a quantidade de enzima produzida por unidade de massa celular no meio de cultura contendo dextrina foi maior do que em meio de cultura contendo amido solúvel, glicose ou lactose. As baixas concentraçöes de carboidrato, a produçäo de enzima por unidade de massa celular em meio contendo glicose foi inferior até emsmo à observada em meio de cultura contendo lactose, onde houve a menor produçäo de alfa-amilase.


Subject(s)
alpha-Amylases , Bacillus subtilis/isolation & purification , Bacillus subtilis/metabolism , Carbon/administration & dosage
19.
Rev. argent. microbiol ; 22(1): 1-6, 1990. ilus
Article in Spanish | LILACS | ID: lil-95046

ABSTRACT

Se estudió el efecto de la privación de triptofano y uracilo sobre la viabilidad del tansformante Bacillus subtilis BSA trp ura y de las cepas parentales Bacillus subtilis PB 168 trp-C y Bacillus subtilis PB 3308 ura. Los ensayos se llevaron a cabo en las condiciones descriptas para el desarrollo de competencia para transformación, durante 16 h. Los resultados obtenidos demostraron que Bacillus subtilis BSA 170 trp ura es resistente a la muerte por carencia de triptofano durante el tiempo del ensayo y a la muerte por carencia de uracilo durante 3h. Luego ocurre una disminución de la recuperación de UFC, acompañada por una reduccción de las absorbancias de los cultivos. La muerte por carencia de uracilo es prevenida en ausencia de triptofano, sugiriendo que reuiere síntesis de proteínas. Las cepas parentales mostraron un comportamiento semejante. Bacillus subtilis PB 168 trp-C mostró ser resistente a la muerte por carencia de triptofano y Bacillus subtilis PB 3308 uta sufrió una reducción de la capacidad para formar colonias, debida a la privación de uracilo, comparable a la de la cepa transformante


Subject(s)
Bacillus subtilis/metabolism , Bacillus subtilis/drug effects , Bacillus subtilis/genetics , Bacterial Proteins/biosynthesis , Tryptophan/pharmacology , Uracil/pharmacology
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