RESUMO
The gastrointestinal microflora regulates the body's functions and plays an important role in its health. Dysbiosis leads to a number of chronic diseases such as diabetes, obesity, inflammation, atherosclerosis, etc. However, these diseases can be prevented by using probiotics living microorganisms that benefit the microflora and, therefore, improve the host organism's health. The most common probiotics include lactic acid bacteria of the Bifidobacterium and Propionibacterium genera. We studied the probiotic properties of the following strains: Bifidobacterium adolescentis ÐС-1909, Bifidobacterium longum infantis ÐС-1912, Propionibacterium jensenii Ð-6085, Propionibacterium freudenreichii Ð-11921, Propionibacterium thoenii Ð-6082, and Propionibacterium acidipropionici Ð-5723. Antimicrobial activity was determined by the 'agar blocks' method against the following test cultures: Escherichia coli ATCC 25922, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa B6643, Proteus vulgaris ATCC 63, and Listeria monocytogenes ATCC 7644. Moderate antimicrobial activity against all the test cultures was registered in Bifidobacterium adolescentis ÐС-1909, Propionibacterium jensenii Ð-6085, and Propionibacterium thoenii Ð-6082. Antioxidant activity was determined by the DPPH inhibition method in all the lactic acid strains. Our study indicated that some Propionibacterium and Bifidobacterium strains or, theoretically, their consortia could be used as probiotic cultures in dietary supplements or functional foods to prevent a number of chronic diseases.
A microbiota gastrointestinal regula as funções do corpo e desempenha um papel importante na sua saúde. A disbiose leva a uma série de doenças crônicas, como diabetes, obesidade, inflamação, aterosclerose, etc. No entanto, essas doenças podem ser prevenidas pelo uso de probióticos − microrganismos vivos que beneficiam a microflora e, portanto, melhoram a saúde do organismo hospedeiro. Os probióticos mais comuns incluem bactérias do ácido láctico dos gêneros Bifidobacterium e Propionibacterium. Nós estudamos as propriedades probióticas das seguintes cepas: Bifidobacterium adolescentis ÐС-1909, Bifidobacterium longum infantis ÐС-1912, Propionibacterium jensenii Ð-6085, Propionibacterium freudenreichii Ð-11921, Propionibacterium thoenii Ð-6082 Ð-6082 acid e Propionibacterium thoenii Ð-6082 Ð-6082 acidibion. A atividade antimicrobiana foi determinada pelo método de 'blocos de ágar' contra as seguintes culturas de teste: Escherichia coli ATCC 25922, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa B6643, Proteus vulgaris ATCC 63 e Listeria monocytogenes moderada atividade ATCC 7644. Uma atividade antimicrobiana moderada contra todas as culturas de teste foi registrado em Bifidobacterium adolescentis ÐС-1909, Propionibacterium jensenii Ð-6085 e Propionibacterium thoenii Ð-6082. A atividade antioxidante foi determinada pelo método de inibição do DPPH em todas as cepas de ácido lático. Nosso estudo indicou que algumas cepas de Propionibacterium e Bifidobacterium − ou, teoricamente, seus consórcios − poderiam ser usadas ââcomo culturas probióticas em suplementos dietéticos ou alimentos funcionais para prevenir uma série de doenças crônicas.
Assuntos
Animais , Propionibacterium , Bifidobacterium , Ácido Láctico , Probióticos , Lactobacillales , Microbioma GastrointestinalRESUMO
Acidipropionibacterium acidipropionici is widely used for many applications, such as propionic acid production, cereal silage, and also as probiotic. Due to this plethora of applications, new isolates of A. acidipropionici with improved features are being searched for. These new isolates must be accurately identified, however, most approaches become expensive and time-consuming when the number of isolates is high. On the contrary, fluorescence in situ hybridization allows the affordable, reliable, and rapid identification of microorganisms in pure cultures and environmental and medical samples. Therefore, the aim of this work was to apply a fluorescent in situ hybridization probe for the reliable identification of new A. acidipropionici isolates. To this end, probe Pap446, specific for A. acidipropionici, was validated by hybridization assays with strains of this species from different origins, other species of the same genus or family, and unrelated genera. Eight isolates with propionibacterium characteristics were obtained from milk and feces of cows. Probe Pap446, hybridized only with isolates III and VI. The identity of these isolates was further confirmed by PCR using group and species-specific primers for propionibacteria and 16S rDNA sequencing.
Assuntos
Propionibacterium , Silagem , Bovinos , Animais , Hibridização in Situ Fluorescente , Propionibacterium/genética , Silagem/microbiologia , Especificidade da EspécieRESUMO
The gastrointestinal microflora regulates the body's functions and plays an important role in its health. Dysbiosis leads to a number of chronic diseases such as diabetes, obesity, inflammation, atherosclerosis, etc. However, these diseases can be prevented by using probiotics - living microorganisms that benefit the microflora and, therefore, improve the host organism's health. The most common probiotics include lactic acid bacteria of the Bifidobacterium and Propionibacterium genera. We studied the probiotic properties of the following strains: Bifidobacterium adolescentis ÐС-1909, Bifidobacterium longum infantis ÐС-1912, Propionibacterium jensenii Ð-6085, Propionibacterium freudenreichii Ð-11921, Propionibacterium thoenii Ð-6082, and Propionibacterium acidipropionici Ð-5723. Antimicrobial activity was determined by the 'agar blocks' method against the following test cultures: Escherichia coli ATCC 25922, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa B6643, Proteus vulgaris ATCC 63, and Listeria monocytogenes ATCC 7644. Moderate antimicrobial activity against all the test cultures was registered in Bifidobacterium adolescentis ÐС-1909, Propionibacterium jensenii Ð-6085, and Propionibacterium thoenii Ð-6082. Antioxidant activity was determined by the DPPH inhibition method in all the lactic acid strains. Our study indicated that some Propionibacterium and Bifidobacterium strains or, theoretically, their consortia could be used as probiotic cultures in dietary supplements or functional foods to prevent a number of chronic diseases.
Assuntos
Anti-Infecciosos , Probióticos , Bifidobacterium , Escherichia coli , Trato Gastrointestinal , Propionibacteriaceae , PropionibacteriumRESUMO
Propionic acid (PA) is a carboxylic acid applied in a variety of processes, such as food and feed preservative, and as a chemical intermediate in the production of polymers, pesticides and drugs. PA production is predominantly performed by petrochemical routes, but environmental issues are making it necessary to use sustainable processes based on renewable materials. PA production by fermentation with the Propionibacterium genus is a promising option in this scenario, due to the ability of this genus to consume a variety of renewable carbon sources with higher productivity than other native microorganisms. However, Propionibacterium fermentation processes present important challenges that must be faced to make this route competitive, such as: a high fermentation time, product inhibition and low PA final titer, which increase the cost of product recovery. This article summarizes the state of the art regarding strategies to improve PA production by fermentation with the Propionibacterium genus. Firstly, strategies associated with environmental fermentation conditions and nutrition requirements are discussed. Subsequently, advantages and disadvantages of various strategies proposed to improve process performance (high cell concentration by immobilization or recycle, co-culture fermentation, genome shuffling, evolutive and metabolic engineering, and in situ recovery) are evaluated.
Assuntos
Embaralhamento de DNA , Propionibacterium , Propionibacterium/genética , Propionibacterium/metabolismo , Fermentação , Propionatos/metabolismoRESUMO
The objective of the present study was to evaluate the use of microbial inoculant on the chemical composition, in vitro gas production, pH, dry matter losses, aerobic stability and microbial population on silages of corn, sorghum and pearl millet in plastic bags silos (without vacuum). The experiment was carried out in a randomized block design, in a 2 × 3 factorial scheme, with and without (control) inoculant consisting of Lactobacillus plantarum and Propionibacterium acidipropionici and on three crops, corn, sorghum and pearl millet, with four replicates. The use of the inoculant did not affect the chemical composition of the silages, except the crude protein (P = 0.0062) and lignin (P = 0.0567) contents. Gas production was neither affected (P > 0.05) by the inoculant nor by the crop. Regarding aerobic stability, we observed that the inoculant affected the temperature of the sorghum silage (P = 0.0123). The inoculant decreased the N-NH3 (P =0.0095) content and increased (P = 0.0441) the lactic acid bacteria population in the silages. Thus, the microbial inoculant did not improve the fermentation profile or nutritional value of corn, pearl millet and sorghum silages in plastic bag silos (without vacuum).(AU)
O objetivo do presente estudo foi avaliar o uso de inoculante bacteriano na composição química, produção de gás in vitro, pH, perdas de matéria seca, estabilidade aeróbia e população microbiana de silagens de milho, sorgo e milheto em silos de sacos plásticos (sem vácuo). O experimento foi conduzido em delineamento de blocos casualizados, em esquema fatorial 2 × 3, [Controle] sem inoculante e Lactobacillus plantarum e Propionibacterium acidipropionici e três culturas, milho, sorgo e milheto, com quatro repetições. O uso do inoculante não afetou a composição química das silagens, exceto proteína bruta (P = 0,0062) e lignina (P = 0,0567). A produção de gás não foi afetada (P > 0,05) pelo inoculante e nem entre as culturas. Na estabilidade aeróbia, observou-se que o inoculante afetou a temperatura da silagem de sorgo (P = 0,0123). O inoculante diminuiu o conteúdo de N-NH3 (P = 0,0095). O inoculante aumentou (P = 0,0441) a população de bactérias ácido-láticas nas silagens. Assim, o inoculante microbiano não melhorou o perfil fermentativo e o valor nutricional das silagens de milho, sorgo e milheto em silos de sacos plásticos (sem vácuo).(AU)
Assuntos
Propionibacterium , Silagem , Zea mays , Pennisetum , Sorghum , Fermentação , Inoculantes Agrícolas , Milhetes , Valor Nutritivo , Técnicas In VitroRESUMO
Vitamin B12 deficiency still persists, mainly caused by low intake of animal food products affecting vegetarians, vegans, and populations of underdeveloped countries. In this study, we investigate the biosynthesis of vitamin B12 by potential probiotic bacterium using an agroindustry residue, the liquid acid protein residue of soybean (LAPRS), as a low-cost, animal derivate-free alternative culture medium. Cultures of Propionibacterium freudenreichii subsp. shermanii ATCC 13673 growing in LAPRS for vitamin B12 biosynthesis were studied using the Plackett-Burman experimental approach, followed by a central composite design 22 to optimize the concentration of significant variables. We also performed a proteolytic treatment of LAPRS and evaluated the optimized-hydrolyzed medium influence on the microbial growth and metabolism in shaker flask and bioreactor experiments. In this all-plant source medium, P. freudenreichii subsp. shermanii produced high concentrations of cells and high amounts of vitamin B12 (0.6 mg/g cells) after process optimization. These results suggest the possibility of producing vitamin B12 by a potential probiotic bacterium in a very cheap, animal derivate-free medium to address the needs of specific population groups, at the same time reducing the production costs of this essential vitamin.
Assuntos
Reatores Biológicos/microbiologia , Meios de Cultura , Propionibacterium/metabolismo , Proteínas de Soja/química , Vitamina B 12/biossíntese , Agricultura , Meios de Cultura/química , Meios de Cultura/metabolismo , Vitamina B 12/análise , Vitamina B 12/químicaRESUMO
The objective of this study was to compare rehydrated corn grain silages using water or whey and inoculated (Lactobacillus plantarum and Propionibacterium acidipropionici) or not. We also verified whether rehydration with whey associated with the bacterial inoculant improves material conservation. The treatments were as follows: silages rehydrated with water without inoculant (SWa); silages rehydrated with water and inoculated (SWaI); silages rehydrated with whey without inoculant (SWe); silages rehydrated with whey and inoculated (SWeI). A completely randomized design was used, with three replications, treatments in a 2 × 2 factorial scheme (RE: rehydration with water or whey, and IN: inoculation or addition of water without chlorine), during the following storage periods (T): 0, 4, 8, 16, 32, and 64 days. There was an RE x IN x T interaction (P < 0.001) for dry matter (DM), with lower values over time for SWe and higher values in SWeI. An interaction between RE x IN (P < 0.001) and IN x T (P < 0.001) was observed for pH, with higher values for SWe at 64 days of storage and a reduction from the first days of ensiling for SWaI and SWeI. The microbiological variables showed an RE x IN x T interaction (P < 0.001), with the highest counts of lactic acid bacteria for SWaI and SWeI up to 8 days of storage and subsequent higher counts in SWa and SWe. Higher counts were obtained in SWe. For the variables of fermentative losses, there was an RE x IN x T interaction (P < 0.001), with SWeI showing lower losses. The inoculation associated with whey for rehydration of corn grain improved the fermentation profile of the silage, with lower pH values and reduced losses.(AU)
O objetivo desse estudo foi comparar as silagens de grão de milho reidratado utilizando água ou soro de leite e inoculados (Lactobacillus plantarum e Propionibacterium acidipropionici) ou não, e verificar se a reidratação com soro associada ao inoculante bacteriano melhora a conservação do material. Os tratamentos foram: silagens reidratadas com água sem inoculante (SA); silagens reidratadas com água com inoculante (SAI); silagens reidratadas com soro de leite inoculante (SS); silagens reidratadas com soro de leite com inoculante (SSI). Foi utilizado um delineamento inteiramente casualizado, com três repetições, tratamentos em esquema fatorial 2 × 2 (RE: reidratação com água ou soro de leite, e IN: inoculação ou adição de água sem cloro), durante os períodos de estocagem (T): 0, 4, 8, 16, 32 e 64dias. Houve interação RE x IN x T (P < 0,001) para variável matéria seca (MS) com menores valores ao longo do tempo para SS e valores mais elevados em SSI. Foi observada interação RE x IN (P < 0,001)e IN x T (P < 0,001) para pH, com maiores valores para SS aos 64 dias de estocagem e redução desde os primeiros dias de ensilagem para SAI e SSI. As variáveis microbiológicas apresentaram interação RE x IN x T (P < 0,001), sendo a maior contagem de bactérias ácido láticas para SAI e SSI até os 8dias de estocagem e após com maiores contagens em SA e SS; maiores contagens foram obtidas em SS. Para as variáveis de perdas fermentativas houve interação RE x IN x T (P < 0,001), em geral, com SSI apresentando menores perdas. A inoculação associada ao uso de soro para reidratação de grão de milho melhorou o perfil fermentativo da silagem, com menores valores de pH e redução das perdas.(AU)
Assuntos
Silagem/análise , Silagem/microbiologia , Soluções para Reidratação/administração & dosagem , Soluções para Reidratação/efeitos adversos , Lactobacillus , PropionibacteriumRESUMO
The objective of this study was to compare rehydrated corn grain silages using water or whey and inoculated (Lactobacillus plantarum and Propionibacterium acidipropionici) or not. We also verified whether rehydration with whey associated with the bacterial inoculant improves material conservation. The treatments were as follows: silages rehydrated with water without inoculant (SWa); silages rehydrated with water and inoculated (SWaI); silages rehydrated with whey without inoculant (SWe); silages rehydrated with whey and inoculated (SWeI). A completely randomized design was used, with three replications, treatments in a 2 × 2 factorial scheme (RE: rehydration with water or whey, and IN: inoculation or addition of water without chlorine), during the following storage periods (T): 0, 4, 8, 16, 32, and 64 days. There was an RE x IN x T interaction (P < 0.001) for dry matter (DM), with lower values over time for SWe and higher values in SWeI. An interaction between RE x IN (P < 0.001) and IN x T (P < 0.001) was observed for pH, with higher values for SWe at 64 days of storage and a reduction from the first days of ensiling for SWaI and SWeI. The microbiological variables showed an RE x IN x T interaction (P < 0.001), with the highest counts of lactic acid bacteria for SWaI and SWeI up to 8 days of storage and subsequent higher counts in SWa and SWe. Higher counts were obtained in SWe. For the variables of fermentative losses, there was an RE x IN x T interaction (P < 0.001), with SWeI showing lower losses. The inoculation associated with whey for rehydration of corn grain improved the fermentation profile of the silage, with lower pH values and reduced losses.
O objetivo desse estudo foi comparar as silagens de grão de milho reidratado utilizando água ou soro de leite e inoculados (Lactobacillus plantarum e Propionibacterium acidipropionici) ou não, e verificar se a reidratação com soro associada ao inoculante bacteriano melhora a conservação do material. Os tratamentos foram: silagens reidratadas com água sem inoculante (SA); silagens reidratadas com água com inoculante (SAI); silagens reidratadas com soro de leite inoculante (SS); silagens reidratadas com soro de leite com inoculante (SSI). Foi utilizado um delineamento inteiramente casualizado, com três repetições, tratamentos em esquema fatorial 2 × 2 (RE: reidratação com água ou soro de leite, e IN: inoculação ou adição de água sem cloro), durante os períodos de estocagem (T): 0, 4, 8, 16, 32 e 64dias. Houve interação RE x IN x T (P < 0,001) para variável matéria seca (MS) com menores valores ao longo do tempo para SS e valores mais elevados em SSI. Foi observada interação RE x IN (P < 0,001)e IN x T (P < 0,001) para pH, com maiores valores para SS aos 64 dias de estocagem e redução desde os primeiros dias de ensilagem para SAI e SSI. As variáveis microbiológicas apresentaram interação RE x IN x T (P < 0,001), sendo a maior contagem de bactérias ácido láticas para SAI e SSI até os 8dias de estocagem e após com maiores contagens em SA e SS; maiores contagens foram obtidas em SS. Para as variáveis de perdas fermentativas houve interação RE x IN x T (P < 0,001), em geral, com SSI apresentando menores perdas. A inoculação associada ao uso de soro para reidratação de grão de milho melhorou o perfil fermentativo da silagem, com menores valores de pH e redução das perdas.
Assuntos
Lactobacillus , Propionibacterium , Silagem/análise , Silagem/microbiologia , Soluções para Reidratação/administração & dosagem , Soluções para Reidratação/efeitos adversosRESUMO
The ban on the use of antibiotics as feed additives for animal growth promotion in the European Union and United States and the expectation of this trend to further expand to other countries in the short term have prompted a surge in probiotic research. Multi-species probiotics including safe and compatible strains with the ability to bind different nutritional lectins with detrimental effects on poultry nutrition could replace antibiotics as feed additives. Lactobacillus salivarius LET201, Lactobacillus reuteri LET210, Enterococcus faecium LET301, Propionibacterium acidipropionici LET103 and Bifidobacterium infantis CRL1395 have proved to be compatible as evaluated through three different approaches: the production and excretion of antimicrobial compounds, growth inhibition by competition for essential nutrients and physical contact, and a combination of both. The safety of P. acidipropionici LET103 was confirmed, since no expression of virulence factors or antibiotic resistance was detected. The innocuity of E. faecium LET301 should be further evaluated, since the presence of genes coding for certain virulence factors (gelE, efaAfm and efaAfs) was observed, albeit no expression of gelE was previously detected for this strain and there are no reports of involvement of efaAfm in animal pathogenicity. Finally, a combination of the five strains effectively protected intestinal epithelial cells of broilers from the cytotoxicity of mixtures of soybean agglutinin, wheat germ agglutinin and concanavalin A. To our knowledge, this is the first time that a combination of strains is evaluated for their protection against lectins that might be simultaneously present in poultry feeds.
Assuntos
Anti-Infecciosos/metabolismo , Bifidobacterium longum subspecies infantis/metabolismo , Enterococcus faecium/metabolismo , Lactobacillus/metabolismo , Doenças das Aves Domésticas/prevenção & controle , Probióticos/farmacologia , Propionibacterium/metabolismo , Animais , Antibiose , Bifidobacterium longum subspecies infantis/genética , Bifidobacterium longum subspecies infantis/crescimento & desenvolvimento , Bifidobacterium longum subspecies infantis/patogenicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Concanavalina A/toxicidade , Farmacorresistência Bacteriana , Enterococcus faecium/genética , Enterococcus faecium/crescimento & desenvolvimento , Enterococcus faecium/patogenicidade , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/fisiologia , Lactobacillus/genética , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/patogenicidade , Lectinas/metabolismo , Modelos Teóricos , Lectinas de Plantas/toxicidade , Probióticos/efeitos adversos , Propionibacterium/genética , Propionibacterium/crescimento & desenvolvimento , Propionibacterium/patogenicidade , Ligação Proteica , Proteínas de Soja/toxicidade , Virulência , Fatores de Virulência/genética , Aglutininas do Germe de Trigo/toxicidadeRESUMO
Cellulose from used toilet paper is a major untapped resource embedded in municipal wastewater which recovery and valorization to valuable products can be optimized. Cellulosic primary sludge (CPS) can be separated by upstream dynamic sieving and anaerobically digested to recover methane as much as 4.02â¯m3/capita·year. On the other hand, optimal acidogenic fermenting conditions of CPS allows the production of targeted short-chain fatty acids (SCFAs) as much as 2.92â¯kg COD/capita·year. Here propionate content can be more than 30% and can optimize the enhanced biological phosphorus removal (EBPR) processes or the higher valuable co-polymer of polyhydroxyalkanoates (PHAs). In this work, first a full set of batch assays were used at three different temperatures (37, 55 and 70⯰C) and three different initial pH (8, 9 and 10) to identify the best conditions for optimizing both the total SCFAs and propionate content from CPS fermentation. Then, the optimal conditions were applied in long term to a Sequencing Batch Fermentation Reactor where the highest propionate production (100-120â¯mg COD/g TVSfed·d) was obtained at 37⯰C and adjusting the feeding pH at 8. This was attributed to the higher hydrolysis efficiency of the cellulosic materials (up to 44%), which increased the selective growth of Propionibacterium acidopropionici in the fermentation broth up to 34%. At the same time, around 88% of the phosphorus released during the acidogenic fermentation was recovered as much as 0.15â¯kg of struvite per capita·year. Finally, the potential market value was preliminary estimated for the recovered materials that can triple over the conventional scenario of biogas recovery in existing municipal wastewater treatment plants.
Assuntos
Celulose/metabolismo , Ácidos Graxos Voláteis/metabolismo , Metano/metabolismo , Fósforo/metabolismo , Esgotos/química , Anaerobiose , Biodegradação Ambiental , Reatores Biológicos/microbiologia , Ácidos Graxos Voláteis/isolamento & purificação , Fermentação , Hidrólise , Metano/isolamento & purificação , Fósforo/isolamento & purificação , Propionibacterium/metabolismo , Águas Residuárias/químicaRESUMO
BACKGROUND: Second-generation ethanol production is a clean bioenergy source with potential to mitigate fossil fuel emissions. The engineering of Saccharomyces cerevisiae for xylose utilization is an essential step towards the production of this biofuel. Though xylose isomerase (XI) is the key enzyme for xylose conversion, almost half of the XI genes are not functional when expressed in S. cerevisiae. To date, protein misfolding is the most plausible hypothesis to explain this phenomenon. RESULTS: This study demonstrated that XI from the bacterium Propionibacterium acidipropionici becomes functional in S. cerevisiae when co-expressed with GroEL-GroES chaperonin complex from Escherichia coli. The developed strain BTY34, harboring the chaperonin complex, is able to efficiently convert xylose to ethanol with a yield of 0.44 g ethanol/g xylose. Furthermore, the BTY34 strain presents a xylose consumption rate similar to those observed for strains carrying the widely used XI from the fungus Orpinomyces sp. In addition, the tetrameric XI structure from P. acidipropionici showed an elevated number of hydrophobic amino acid residues on the surface of protein when compared to XI commonly expressed in S. cerevisiae. CONCLUSIONS: Based on our results, we elaborate an extensive discussion concerning the uncertainties that surround heterologous expression of xylose isomerases in S. cerevisiae. Probably, a correct folding promoted by GroEL-GroES could solve some issues regarding a limited or absent XI activity in S. cerevisiae. The strains developed in this work have promising industrial characteristics, and the designed strategy could be an interesting approach to overcome the non-functionality of bacterial protein expression in yeasts.
Assuntos
Aldose-Cetose Isomerases/metabolismo , Chaperonina 60/genética , Proteínas de Escherichia coli/genética , Proteínas de Choque Térmico/genética , Engenharia de Proteínas/métodos , Saccharomyces cerevisiae/genética , Aldose-Cetose Isomerases/química , Aldose-Cetose Isomerases/genética , Chaperonina 60/metabolismo , Proteínas de Escherichia coli/metabolismo , Etanol/metabolismo , Proteínas de Choque Térmico/metabolismo , Modelos Moleculares , Propionibacterium/enzimologia , Conformação Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/metabolismo , Xilose/metabolismoRESUMO
The increasing demand for propionic acid (PA) production and its wide applications in several industries, especially the food industry (as a preservative and satiety inducer), have led to studies on the low-cost biosynthesis of this acid. This paper gives an overview of the biotechnological aspects of PA production and introduces Propionibacterium as the most popular organism for PA production. Moreover, all process variables influencing the production yield, different simple and complex carbon sources, the metabolic pathway of production, engineered mutants with increased productivity, and modified tolerance against high concentrations of acid have been described. Furthermore, possible methods of extraction and analysis of this organic acid, several applied bioreactors, and different culture systems and substrates are introduced. It can be concluded that maximum biomass and PA production may be achieved using metabolically engineered microorganisms and analyzing the most significant factors influencing yield. To date, the maximum reported yield for PA production is 0.973 g·g-1, obtained from Propionibacterium acidipropionici in a three-electrode amperometric culture system in medium containing 0.4 mM cobalt sepulchrate. In addition, the best promising substrate for PA bioproduction may be achieved using glycerol as a carbon source in an extractive continuous fermentation. Simultaneous production of PA and vitamin B12 is suggested, and finally, the limitations of and strategies for competitive microbial production with respect to chemical process from an economical point of view are proposed and presented. Finally, some future trends for bioproduction of PA are suggested.
Assuntos
Propionatos/metabolismo , Propionibacterium/metabolismo , Propionatos/química , Vitamina B 12/biossíntese , Carbono/metabolismo , Reatores Biológicos , Ácidos Graxos Voláteis/metabolismo , Fermentação , Concentração de Íons de Hidrogênio , Nitrogênio/metabolismoRESUMO
Propionic acid and its derivatives are considered "Generally Recognized As Safe" food additives and are generally used as an anti-microbial and anti-inflammatory agent, herbicide, and artificial flavor in diverse industrial applications. It is produced via biological pathways using Propionibacterium and some anaerobic bacteria. However, its commercial chemical synthesis from the petroleum-based feedstock is the conventional production process bit results in some environmental issues. Novel biological approaches using microorganisms and renewable biomass have attracted considerable recent attention due to economic advantages as well as great adaptation with the green technology. This review provides a comprehensive overview of important biotechnological aspects of propionic acid production using recent technologies such as employment of co-culture, genetic and metabolic engineering, immobilization technique and efficient bioreactor systems.
Assuntos
Reatores Biológicos/microbiologia , Engenharia Metabólica/métodos , Propionatos/metabolismo , Propionibacterium/metabolismo , Células ImobilizadasRESUMO
Diariamente los seres humanos están en interacción con objetos de uso continuo, como el papel moneda, sin el conocimiento de que estos almacenan microorganismos y de que nos exponemos al contacto con potenciales patógenos. La composición de la comunidad bacteriana en un billete colombiano fue determinada mediante el secuenciamiento profundo de librerías de amplicones 16S. Se encontraron 233 géneros bacterianos; 12 de estos géneros corresponden a especies con potencial patogénico. El género más abundante fue Propionibacterium, seguido de Streptococcus, Staphylococcus Pseudomonas . Este es el primer reporte de la diversidad bacteriana que puede ser alojada en este objeto de alta circulación en Colombia. Pocos estudios en el mundo han mostrado este nivel de detalle de la microbiota en billetes de circulación y ofrece un panorama mucho más amplio de la exposición diaria a microorganismos al utilizar papel moneda en las condiciones en las que se utiliza en Colombia.
Commonly used objects such as currency paper can be colonised by bacteria and can serve as carriers of microbes. This colonisation might expose us to unnoticed pathogenic bacteria. In this study, the researchers obtained a detailed panorama of the microbes that can be carried on currency notes in Colombia by using 454 next-generation deep sequencing of 16S amplicón libraries. A total of 233 bacterial genera were detected and classified, 12 of which are potential human pathogens. The most abundant genera were Propionibacterium, Streptococcus, Staphylococcus and Pseudomonas. To date, this is the first in-depth analysis of the microbiota carried by circulating banknotes in our continent and it offers insights into daily exposure to microbes when using banknotes in Colombia.
Assuntos
Humanos , Masculino , Feminino , Papel , Bactérias , Microbiota , Propionibacterium , Streptococcus , Saúde Ambiental , Colômbia , NoxasRESUMO
Different studies in animal rearing claim the probiotic potential of species of the genus Propionibacterium. The effects of strains of Propionibacterium acidipropionici isolated from poultry intestine on microbiota activity and intestinal mucosa development were investigated in the early stage of rearing chicks and the safety of the dose used was investigated. The strains P. acidipropionici LET105 and LET107, administered as monoculture to chicks from the 1st to 14th day of life in a daily dose of 106 cfu/ml administered in the drinking water resulted harmless. The animals arrived at the expected weight for age and no differences were observed with respect to the food intake and water consumption related to control without bacteria administration. The analysis of microbiota composition revealed the presence of propionibacteria at the middle and end of the trial only in treated groups. Normal development of lactic acid bacteria and bifidobacteria, and slow colonisation by Bacteroides at the 7th day of the study was observed in the same groups. Analysis of the organic acids concentrations in the caecal content of birds revealed higher lactic acid and lower butyric acid production. Lower short chain fatty acids total concentration than expected during treatment was related to a better development of the gut mucosa. Increase in length of villus-crypt units, goblet cells counts and neutral mucins production were evidenced. Higher mucus secretion produced by dietary supplementation with propionibacteria could provide increased protection against pathogens.
Assuntos
Galinhas/microbiologia , Microbioma Gastrointestinal , Probióticos/farmacologia , Propionibacterium , Ração Animal , Animais , Butiratos/metabolismo , Ceco/imunologia , Ceco/microbiologia , Galinhas/crescimento & desenvolvimento , Galinhas/fisiologia , Dieta/veterinária , Ácidos Graxos Voláteis/metabolismo , Feminino , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Intestinos/imunologia , Intestinos/microbiologia , Ácido Láctico/metabolismo , Distribuição Aleatória , Aumento de Peso/efeitos dos fármacosRESUMO
Adhesion to the host intestinal mucosa is considered relevant for orally delivered probiotics as it prolongs their persistence in the gut and their health promoting effects. Classical propionibacteria are microorganisms of interest due to their role as dairy starters as well as for their functions as probiotics. Propionibacterium acidipropionici Q4, is a dairy strain isolated from a Swiss-type cheese made in Argentina that displays probiotic potential. In the present work we assessed the ability of this strain to adhere to the human enterocyte-like HT-29 cell line and to counteract the adhesion of two common human enteropathogens, such as Escherichia coli C3 and Salmonella Enteritidis 90/390. The results were compared with those obtained with the well-known probiotic Lactobacillus rhamnosus GG. P. acidipropionici Q4 showed a high adhesion capacity, even higher than the reference strain L. rhamnosus GG (42.3±4.4% and 36.2±2.3%, respectively), whereas adhesion of enteropathogens was significantly lower (25.2±2.2% for E. coli and 21.0±3.4% for S. Enteritidis). Propionibacteria as well as lactobacilli were able to inhibit by exclusion and competition the adherence of E. coli C3 and S. Enteritidis 90/390 whereas only L. rhamnosus GG displaced S. Enteritidis from HT-29 intestinal cells. Inhibition of pathogens by propionibacteria was not exerted by antimicrobials or coaggregation but was mainly due to exclusion by cell surface components, such as proteins and carbohydrates. The relevance of cell surface proteins (CSP) for preventing pathogens infection was confirmed by their concentration dependent effect observed for both pathogens: 100 µg/ml of CSP inhibited E. coli attachment almost as untreated propionibacteria, whereas it partially inhibited the attachment of S. Enteritidis. Results suggest that P. acidipropionici Q4 could be considered for the development of propionibacteria containing functional foods helpful in counteracting enteropathogen infection.
Assuntos
Antibiose , Aderência Bacteriana , Enterócitos/microbiologia , Escherichia coli/fisiologia , Propionibacterium/fisiologia , Salmonella enteritidis/fisiologia , Células HT29 , Humanos , Lacticaseibacillus rhamnosus/fisiologiaRESUMO
AIMS: Propionibacterium freudenreichii is an actinobacterium widely used in dairy industry during the ripening process of Swiss-type cheeses and which presents probiotic properties. P. freudenreichii is reportedly a hardy bacterium, able to survive during the cheese-making process and when subjected to digestive stresses. During this study the long-term survival (LTS) of P. freudenreichii was investigated for 11 days by means of phenotypic characterization in a culture medium without the addition of any nutrients. METHODS AND RESULTS: For 11 days, in a non-nutrient supplemented culture medium, eight strains were monitored by measuring their optical density, counting colony-forming units (CFU) and using LIVE/DEAD staining and microscopy observation. Under these conditions, all strains displayed high survival rates in the culture medium, their culturability reaching more than 9 log10 CFU ml(-1) after 2 days. After 11 days, this value ranged from 7·8 to 8·2 log10 CFU ml(-1) depending on the strain, and at least 50% of the P. freudenreichii population displayed an intact envelope. As lysis of part of a bacterial population may be a microbial strategy to recover nutrients, in CIRM-BIA 138 (the strain with the highest population at day 11), cell lysis was assessed by quantifying intact bacterial cells using qPCR targeting the housekeeping gene tuf. No lysis was observed. CONCLUSION: Taken together, our results suggest that P. freudenreichii strains use a viable but nonculturable state to adapt to the LTS phase. SIGNIFICANCE AND IMPACT OF THE STUDY: Assessing the viability of P. freudenreichii and understanding their mechanisms for survival should be of great interest regarding their potential probiotic applications.
Assuntos
Meios de Cultura/metabolismo , Propionibacterium/crescimento & desenvolvimento , Meios de Cultura/análise , Viabilidade Microbiana , Propionibacterium/metabolismoRESUMO
Dairy propionibacteria are used as ripening cultures for the production of Swiss-type cheeses, and some strains have potential for use as probiotics. This study investigated the biodiversity of wild dairy Propionibacteria isolates in dairy farms that produce Swiss-type cheeses in Minas Gerais State, Brazil. RAPD and PFGE were used for molecular typing of strains and MLST was applied for phylogenetic analysis of strains of Propionibacterium freudenreichii. The results showed considerable genetic diversity of the wild dairy propionibacteria, since three of the main species were observed to be randomly distributed among the samples collected from different farms in different biotopes (raw milk, sillage, soil and pasture). Isolates from different farms showed distinct genetic profiles, suggesting that each location represented a specific niche. Furthermore, the STs identified for the strains of P. freudenreichii by MLST were not related to any specific origin. The environment of dairy farms and milk production proved to be a reservoir for Propionibacterium strains, which are important for future use as possible starter cultures or probiotics, as well as in the study of prevention of cheese defects.
Assuntos
Biodiversidade , Microbiologia de Alimentos , Propionibacterium/genética , Animais , Brasil , Queijo/microbiologia , Microbiologia Ambiental , Variação Genética , Leite/microbiologia , Tipagem de Sequências Multilocus , Filogenia , Probióticos/isolamento & purificação , Propionibacterium/classificação , Propionibacterium/isolamento & purificação , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
Free fatty acids are important flavor compounds in cheese. Propionibacterium freudenreichii is the main agent of their release through lipolysis in Swiss cheese. Our aim was to identify the esterase(s) involved in lipolysis by P. freudenreichii. We targeted two previously identified esterases: one secreted esterase, PF#279, and one putative cell wall-anchored esterase, PF#774. To evaluate their role in lipolysis, we constructed overexpression and knockout mutants of P. freudenreichii CIRM-BIA1(T) for each corresponding gene. The sequences of both genes were also compared in 21 wild-type strains. All strains were assessed for their lipolytic activity on milk fat. The lipolytic activity observed matched data previously reported in cheese, thus validating the relevance of the method used. The mutants overexpressing PF#279 or PF#774 released four times more fatty acids than the wild-type strain, demonstrating that both enzymes are lipolytic esterases. However, inactivation of the pf279 gene induced a 75% reduction in the lipolytic activity compared to that of the wild-type strain, whereas inactivation of the pf774 gene did not modify the phenotype. Two of the 21 wild-type strains tested did not display any detectable lipolytic activity. Interestingly, these two strains exhibited the same single-nucleotide deletion at the beginning of the pf279 gene sequence, leading to a premature stop codon, whereas they harbored a pf774 gene highly similar to that of the other strains. Taken together, these results clearly demonstrate that PF#279 is the main lipolytic esterase in P. freudenreichii and a key agent of Swiss cheese lipolysis.
Assuntos
Queijo/microbiologia , Esterases/metabolismo , Lipólise , Propionibacterium/enzimologia , Esterases/genética , Microbiologia de Alimentos , Técnicas de Inativação de Genes , Variação Genética , Dados de Sequência Molecular , Propionibacterium/genéticaRESUMO
The prevention and control of pathogens colonization through probiotics administration in poultry feeding is of increasing interest. The genus Propionibacterium is an attractive candidate for the development of probiotic cultures as they produce short chain fatty acids (SCFA) by carbohydrates fermentation. The presence of strains of this genus in hens of conventional production systems and backyard hens was investigated. Propionibacteria were isolated from the intestine and identified by physiological and biochemical tests. PCR amplification of the 16S rRNA gene of the isolates was performed and products were compared with sequences from databases. The presence of the genus Propionibacterium was demonstrated in 26% of hens and Propionibacterium acidipropionici and Propionibacterium avidum were the identified species. A comparative study of their physiological and functional characteristics was performed. P. acidipropionici strains were the most resistant to in vitro gastrointestinal digestion, but the adhesion to intestinal tissue was strain dependent. Some differences were found between both species with respect to their growth and SCFA production in an in vitro cecal water model, but all the strains were metabolically active. The production of SCFA in cecal slurries inoculated with the strain P. acidipropionici LET 105 was 30% higher than in non-inoculated samples. SCFA concentrations obtained were high enough to inhibit Salmonella enterica serovar Enteritidis when assayed in a cecal water model. P. acidipropionici LET 105 was also able to compete with Salmonella for adhesion sites on the intestinal mucosa in ex vivo assays. Results contribute to the knowledge of the species diversity of the genus Propionibacterium in the intestine of poultry and provide evidence of their potential for probiotics products development.