RESUMEN
This study investigated the effect of cooking on the levels of 3-chloro-1, 2-propanediol esters (3-MCPDEs), 2-chloro-1, 3-propanediol esters (2-MCPDEs) and glycidyl esters (GEs) in deep-fried rice cracker, fried potato, croquette, fish fillet, chicken fillet and cooking oils (rice bran oil and palm oil). The levels of 2-/3-MCPDE in rice cracker fried with rice bran oil and the used oil remained about the same, while the levels of GEs in them fell with frying time. The levels of 2-/3-MCPDEs in fried potato, croquette, fried fish and chicken cutlet fried with rice bran oil and palm oil respectively fell with frying time, while the level of GEs in them remained about the same. The levels of 2-/3-MCPDEs and GEs in fried rice cooked with rice bran oil were under the method limit of quantification. These results provide insights the cooking has no influence with the levels of 2-/3-MCPDEs and GEs in cooked foods.
Asunto(s)
Culinaria , Ésteres , Calor , Aceite de Palma , Aceite de Salvado de Arroz , alfa-Clorhidrina , Culinaria/métodos , Ésteres/análisis , Aceite de Palma/química , Aceite de Salvado de Arroz/química , alfa-Clorhidrina/análisis , Ácidos Grasos/análisis , Aceites de Plantas/química , Análisis de los Alimentos , Animales , Factores de Tiempo , Glicoles de Propileno/análisis , Compuestos Epoxi/análisis , Grasas de la Dieta/análisis , Pollos , Alimentos ProcesadosRESUMEN
In this study, 241 vegetable-oil food samples were collected from the Hangzhou market in China and analysed for fatty acid esters of 3- and 2-monochloropropanediol (3-MCPD and 2-MCPD) using non-derivative gas chromatography tandem mass spectrometry (GC-MS/MS). Food consumption data were taken from a food consumption survey of urban and rural residents in Hangzhou city performed in 2010-2011. Levels of 3-MCPD esters in edible oil ranged from not detected to 7.98 mg/kg, and the highest mean levels were found in tea seed oil, with concentrations of 2.94 mg/kg. Esters of 2-MCPD levels ranged from not detected to 4.03 mg/kg, and the highest mean levels were also found in tea seed oil, containing 1.49 mg/kg. The range of mean dietary intake of 3-MCPD esters in different groups of edible oil was from 0.096 to 1.54 µg/kg body weight (bw) per day, which is lower than the tolerable daily intake (TDI) established by the European Food Safety Authority (EFSA) (2 µg/kg bw/day). For people aged above 6 years old, the dietary intake of 3-MCPD from edible oil was 0.42 µg/kg bw per day (mean) and 1.22 µg/kg bw per day (P97.5). The range of mean dietary intake of 2-MCPD esters in different groups of edible oil was from 0.025 to 0.79 µg/kg bw/day, and 2-MCPD esters intake was 0.20 µg/kg bw per day (mean) and 0.60 µg/kg bw per day (P97.5). In addition, the dietary intake exposure to 3-MCPD and 2-MCPD esters for urban residents was lower than that for rural residents. The findings indicate that the potential health risks caused by dietary 3-MCPD esters from edible oils were of low concern for most of the Hangzhou residents. However, the exposure risk for consumers with excessive consumption of certain kind of edible oil calls for attention.
Asunto(s)
Aceites de Plantas/química , Glicoles de Propileno/química , China , Exposición Dietética , Análisis de los Alimentos , Contaminación de Alimentos , Humanos , Glicoles de Propileno/análisis , Medición de RiesgoRESUMEN
OBJECTIVES: To select a microbial consortium from intertidal sludge and evaluate its ability to convert crude glycerol from biodisel to high value-added products such as 1,3-propanediol (1,3-PDO) and lactic acid (LA). RESULTS: A microbial consortium named CJD-S was selected from intertidal sludge and exhibited excellent performance for the conversion of crude glycerol to 1,3-PDO and LA. The composition of CJD-S was determined to be 85.99% Enterobacteriaceae and 13.75% Enterococcaceae by 16S rRNA gene amplicon high-throughput sequencing. In fed-batch fermentation with crude glycerol under nonsterile conditions, the highest concentrations of 1,3-PDO and LA were 41.47 g/L and 45.86 g/L, respectively. CONCLUSIONS: The selected microbial consortium, CJD-S, effectively converted crude glycerol to 1,3-PDO and LA under nonsterile conditions and can contribute to the sustainable development of the biodiesel industry.
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Glicerol/metabolismo , Ácido Láctico/metabolismo , Consorcios Microbianos/fisiología , Glicoles de Propileno/metabolismo , Aguas del Alcantarillado/microbiología , Biocombustibles , Reactores Biológicos , Fermentación , Ácido Láctico/análisis , Glicoles de Propileno/análisisRESUMEN
A new atmospheric pressure ionization method, plasmaspray ionization, termed as PSI, was developed to be an alternative ambient ion source for mass spectrometry. It comprises a plasma jet device and a sample spray part. While the nonthermal plasma jet strikes the surface of stainless steel tube out of the spray capillary, the sprayed sample will be ionized with the assistant of auxiliary gas. Although PSI is a little bit more complex than electrospray ionization (ESI) in instrument, it shows both better linearity and higher sensitivity for organic compounds. For protein samples, it presents wider distributions of multiply charged ions and higher mass resolution without sacrificing any sensitivity. For the mechanism of PSI, the charge build-up process on the tip of capillary should play a key role for the ion formation, and the stimulated pulsed voltage on the flow tube will promote the ion aggregation speed until the charge density is high enough. PSI source contains the features of plasma ionization and ESI and can be considered as a novel combo bridging these techniques. These results reflect that this method of PSI can be applied and further developed as a versatile new ion source for a wild range of organic and biological samples.
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Espectrometría de Masas/instrumentación , Espectrometría de Masas/métodos , Compuestos Orgánicos/análisis , Proteínas/análisis , Ionización del Aire , Presión Atmosférica , Cafeína/análisis , Lecitinas/análisis , Polímeros/análisis , Glicoles de Propileno/análisis , Reserpina/análisisRESUMEN
Dropropizine is a peripheral antitussive drug that acts by inhibiting cough reflex through its action on the peripheral receptors and their afferent conductors. It is marketed in a racemic form or its pure enantiomer called levodropropizine and both are available worldwide in various drug dosage formulations such as tablets, sirup and oral solution. Due to the widespread use of antitussives in the clinic it is necessary to develop efficient analytical methodologies for quality control and also for pharmacokinetic, bioavailability and bioequivalence studies. This review presents a survey of the characteristics, properties and analytical methods used for drug determination, being carried out through scientific articles as well as in official compendia. From the analyzed studies, the majority reports the use of HPLC/UV techniques for drug determination, but also spectrophotometric UV/Vis methods as well as gas chromatography, and voltammetric, potentiometric and conductometric titration methods. In addition, the methodologies addressed the determination of dropropizine or levodropropizine in different types of matrices such as raw material, pharmaceutical formulations, plasma and urine. Despite the extensive clinical use of dropropizine, data from this review evidenced a still limited number of studies dealing with analytical methods for its determination in different matrices, which may be of concern since the applicability of these methods is important for quality assurance, efficacy and safety of the medicine.
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Antitusígenos/análisis , Cromatografía Líquida de Alta Presión/métodos , Glicoles de Propileno/análisis , Antitusígenos/farmacocinética , Antitusígenos/uso terapéutico , Tos/tratamiento farmacológico , Cromatografía de Gases y Espectrometría de Masas , Semivida , Humanos , Glicoles de Propileno/farmacocinética , Glicoles de Propileno/uso terapéutico , Espectrofotometría , Estereoisomerismo , Comprimidos/químicaRESUMEN
With the aggravation of environmental pollution and energy crisis, the sustainable microbial fermentation process of converting glycerol to 1,3-propanediol (1,3-PDO) has become an attractive alternative. However, the difficulty in the online measurement of glycerol and 1,3-PDO creates a barrier to the fermentation process and then leads to the residual glycerol and therefore, its wastage. Thus, in the present study, the four-input artificial neural network (ANN) model was developed successfully to predict the concentration of glycerol, 1,3-PDO, and biomass with high accuracy. Moreover, an ANN model combined with a kinetic model was also successfully developed to simulate the fed-batch fermentation process accurately. Hence, a soft sensor from the ANN model based on NaOH-related parameters has been successfully developed which cannot only be applied in software to solve the difficulty of glycerol and 1,3-PDO online measurement during the industrialization process, but also offer insight and reference for similar fermentation processes.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Clostridium butyricum/metabolismo , Fermentación/fisiología , Redes Neurales de la Computación , Glicoles de Propileno , Reactores Biológicos/microbiología , Medios de Cultivo/análisis , Medios de Cultivo/química , Medios de Cultivo/metabolismo , Glicerol/análisis , Glicerol/metabolismo , Cinética , Glicoles de Propileno/análisis , Glicoles de Propileno/metabolismoRESUMEN
Native electrospray mass spectrometry is a powerful method for determining the native stoichiometry of many polydisperse multi-subunit biological complexes, including multi-subunit protein complexes and lipid-bound transmembrane proteins. However, when polydispersity results from incorporation of multiple copies of two or more different subunits, it can be difficult to analyze subunit stoichiometry using conventional mass spectrometry analysis methods, especially when m/z distributions for different charge states overlap in the mass spectrum. It was recently demonstrated by Marty and co-workers (K. K. Hoi, et al., Anal. Chem., 2016, 88, 6199-6204) that Fourier Transform (FT)-based methods can determine the bulk average lipid composition of protein-lipid Nanodiscs assembled with two different lipids, but a detailed statistical description of the composition of more general polydisperse two-subunit populations is still difficult to achieve. This results from the vast number of ways in which the two types of subunit can be distributed within the analyte ensemble. Here, we present a theoretical description of three common classes of heterogeneity for mixed-subunit analytes and demonstrate how to differentiate and analyze them using mass spectrometry and FT methods. First, we first describe FT-based analysis of mass spectra corresponding to simple superpositions, convolutions, and multinomial distributions for two or more different subunit types using model data sets. We then apply these principles with real samples, including mixtures of single-lipid Nanodiscs in the same solution (superposition), mixed-lipid Nanodiscs and copolymers (convolutions), and isotope distribution for ubiquitin (multinomial distribution). This classification scheme and the FT method used to study these analyte classes should be broadly useful in mass spectrometry as well as other techniques where overlapping, periodic signals arising from analyte mixtures are common.
Asunto(s)
Proteínas de la Membrana/análisis , Polietilenglicoles/análisis , Glicoles de Propileno/análisis , Subunidades de Proteína/análisis , 1,2-Dipalmitoilfosfatidilcolina/química , Análisis de Fourier , Espectrometría de Masas/métodos , Proteínas de la Membrana/química , Nanoestructuras/química , Fosfatidilcolinas/química , Polietilenglicoles/química , Glicoles de Propileno/química , Subunidades de Proteína/químicaRESUMEN
AIM: In this investigation, a dye-based pH-stat method was devised for monitoring steady production of 1,3-propanediol (1,3-PDO) during bioprospecting of glycerol-assimilating soil microbiome. METHODS AND RESULTS: Soil samples were collected from two potential sites of CSIR-IIP, India. Selective enrichment of microbial consortia was done using the glycerol-based medium at initial stage, followed by purification to isolated colonies, after positive high-performance liquid chromatography detection of 1,3-PDO in the fermentation broth. When the purified isolated were re-tested for 1,3-PDO production, only two isolates namely Isolate 1 and Isolate 3 were capable of producing the targeted product preferably under anaerobic conditions. Based on better 1,3-PDO fermentation efficiency (Isolate 3, 22% vs Isolate 1, 4·48%) and acetic acid as the only major by-product, Isolate 3 was shortlisted for further studies. A dye-based technique was devised in which bromothymol blue was incorporated into the medium to monitor the pH drop due to acetic acid formation and hence change in colour. Visual change in colour helped in intermittent pH restoration. During fermentation, with pH stat being 8-8·5, Isolate 3 at 32°C yielded 0·67 mol mol-1 1,3-PDO within a short span of 12 h only with an initial concentration of glycerol being 20 g l-1 . Phylogenetic analysis revealed that Isolate 3 shared 95·8% homology with Citrobacter freundii CFNIH1 and hence designated as C. freundii IIP DR3. CONCLUSION: This study demonstrated that during bioprospecting glycerol-assimilating microbiome, dye-based technique can be successfully employed. This technique can further be exploited to monitor consistent production of all microbial secondary metabolites that accompanies acid production. SIGNIFICANCE AND IMPACT OF THE STUDY: Incorporation of 'Bromothymol blue' can visually help in the identification of pH drop in the medium, so that pH stat can be easily maintained during 1,3-PDO production from glycerol especially under shake flask conditions.
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Bacterias/metabolismo , Glicerol/metabolismo , Glicoles de Propileno/metabolismo , Microbiología del Suelo , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bioprospección , Fermentación , Concentración de Iones de Hidrógeno , India , Consorcios Microbianos , Filogenia , Glicoles de Propileno/análisis , Suelo/químicaRESUMEN
The production of 1,3-propanediol (1,3-PDO) is an important fermentation process. However, 1,3-PDO could not be distinguished separately and efficiently in fermentations previously because it has a highly similar molecular structure to the feedstock glycerol (GLY) and by-product lactic acid (Lac), which leads to the difficulty of quantification. In this paper, a low-cost and environmentally friendly biosensor based on surface-enhanced Raman scattering (SERS) technique was developed. Using it, the concentration of 1,3-PDO and Lac in a fermentation solution can be determined directly from their respective characteristic peaks in Raman spectroscopy. Moreover, by analyzing the respective contributions of 1,3-PDO, Lac, and GLY to the integrated intensities of the 2920â¯cm-1 Raman peak common to these three substances, the concentration of GLY could also be quantified. SERS study on various 1,3-PDO:GLY and Lac:GLY molar ratios were conducted to establish the proportional relationships of these compounds by analyzing the relationship between the concentration and the Raman peak intensities. The 1,3-PDO:Lac:GLY with serial concentration gradient was carried out to verify the relationship between the concentration and the Raman peak intensities by the high-performance liquid chromatography (HPLC) with relative deviations <25%. Concentrations of 1,3-PDO and Lac as low as 1â¯g/L and concentration of GLY as low as 4â¯g/L were analyzed to determine the limit of detection. Therefore, this new method allows the rapid quantification of 1,3-PDO, Lac and GLY concentrations on a SERS-based biosensor.
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Técnicas Biosensibles , Glicoles de Propileno/análisis , Espectrometría Raman/métodos , Cromatografía Líquida de Alta Presión , Fermentación , Glicerol/análisis , Glicerol/metabolismo , Ácido Láctico/análisis , Ácido Láctico/metabolismo , Límite de Detección , Glicoles de Propileno/metabolismo , Espectrometría Raman/instrumentaciónRESUMEN
Synthetic biology emerges as a powerful approach for unlocking the potential of cyanobacteria to produce various chemicals. However, the highly oxidative intracellular environment of cyanobacteria is incompatible to numerous introduced enzymes from anaerobes. In this study, we explore a strategy based on natural compartmentalization of cyanobacterial heterocysts to overcome the incompatibility. Hence, the oxygen-sensitive 1,3-propanediol (1,3-PDO) biosynthetic pathway was selected as a model and insulated in heterocysts to evaluate the proposed strategy. Thus, the genes from different sources for 1,3-PDO production were tandemly arrayed with promoter, resulting the assembled 1,3-PDO synthetic cassettes. Then the synthetic cassettes were integrated into the chromosome of Anabaena sp. strain PCC7120 by homologous recombination, respectively. The engineered strain P11 containing the genes from facultative anaerobe Klebsiella pneumoniae (cassette KP) accumulated 46.0 mg L-1 of 1,3-PDO when heterocysts were present, which is approximately 1.7-fold higher than that of no heterocysts. As for the strains (P12, P13, and P14) containing the genes from strictly anaerobic bacterium Clostridium butyricum (cassette CB), the product 1,3-PDO could only be detected when heterocysts were present. These results indicate that insulation of the oxygen-sensitive 1,3-PDO pathway with heterocysts is an effective way to protect these enzymes in cyanobacteria. The strategy may have the potential of serving as a universal strategy to overcome the incompatibility of oxygen-sensitivity in synthetic biology.
Asunto(s)
Anabaena/genética , Klebsiella pneumoniae/genética , Luz , Glicoles de Propileno/metabolismo , Proteínas Bacterianas/genética , Dióxido de Carbono/metabolismo , Cromosomas Bacterianos/genética , Clostridium butyricum/genética , Cromatografía de Gases y Espectrometría de Masas , Recombinación Homóloga , Hidroliasas/genética , Ingeniería Metabólica/métodos , Monoéster Fosfórico Hidrolasas/genética , Plásmidos/genética , Plásmidos/metabolismo , Glicoles de Propileno/análisis , Glicoles de Propileno/químicaRESUMEN
A sensitive LC-MS/MS method was established to quantify diols and glycerol in fermentation broth using trichloroacetyl isocyanate as instantaneous derivatization reagent for monitoring the production of 1,3-propanediol and 2,3-butanediol from the biodiesel biorefinery process. Due to the derivatization reaction was very quickly at room temperature, only 1â¯min was needed for the reaction process. In addition, both extraction of analytes and evaporation of water were not employed in the analytical procedure. Furthermore, the isotope of chlorine was beneficial for understanding of the secondary mass spectrum and avoiding false positive results. Therefore, much more accurate results of diols and glycerol concentration in fermentation could be obtained even at very low levels for the evaluation of microbial metabolism pathway modification.
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Alcoholes/análisis , Técnicas de Química Analítica/métodos , Cromatografía Liquida , Fermentación , Glicerol/análisis , Isocianatos/química , Espectrometría de Masas en Tándem , Biocombustibles , Butileno Glicoles/análisis , Glicoles de Propileno/análisisRESUMEN
A novel method has been developed for the direct, sensitive, and rapid detection of bronopol in rice using a simple solid-phase extraction (SPE) procedure followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), with electrospray ionization (ESI). Bronopol was stable under acidic conditions, and an acidic environment was thus needed before sample loading to ensure the stability of bronopol. Rice extracts containing bronopol were pretreated using a hydrophilic-lipophilic balanced (Bond Elut Plexa) cartridge to reduce the matrix effect. An XDB-C18 column (150 mm × 2.1 mm, 3.5 µm) was used for chromatographic separations, with a mobile phase comprising methanol and aqueous ammonium formate (5 mM). The linearity of the method was satisfactory with regression coefficient (R 2) = 0.9992. The limit of quantification was 3.3 µg kg-1. Three spiked levels (25, 125 and 625 µg kg-1) were used to determine the recovery of bronopol, which was found to be 73.3-96.7%, with relative standard deviations (RSD) in the range 1.2-7.9%. The RSD for intra-day precision (n = 7) was 7.6% and the RSD for inter-day precision (n = 15) was 8.3%. The newly developed analytical method was successfully used to quantify bronopol in rice samples.
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Residuos de Medicamentos/análisis , Oryza/química , Glicoles de Propileno/análisis , Extracción en Fase Sólida/métodos , Espectrometría de Masas en Tándem/métodos , Cromatografía Líquida de Alta Presión/métodos , Límite de Detección , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de Electrospray/métodosRESUMEN
We utilized a photoautotrophic organism to synthesize 1,2-propanediol from carbon dioxide and water fueled by light. A synthetic pathway comprising mgsA (methylglyoxal synthase), yqhD (aldehyde reductase), and adh (alcohol dehydrogenase) was inserted into Synechocystis sp. PCC6803 to convert dihydroxyacetone phosphate to methylglyoxal, which is subsequently reduced to acetol and then to 1,2-propanediol. 1,2-propanediol could be successfully produced by Synechocystis, at an approximate rate of 55 µmol h-1 gCDW-1 . Surprisingly, maximal productivity was observed in the stationary phase. The production of 1,2-propanediol was clearly coupled to the turn-over of intracellular glycogen. Upon depletion of the glycogen pool, product formation stopped. Reducing the carbon flux to glycogen significantly decreased final product titers. Optimization of cultivation conditions allowed final product titers of almost 1 g L-1 (12 mM), which belongs to the highest values published so far for photoautotrophic production of this compound.
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Glucógeno/metabolismo , Glicoles de Propileno/metabolismo , Synechocystis/metabolismo , Procesos Autotróficos , Biotecnología , Dióxido de Carbono/metabolismo , Glicoles de Propileno/análisis , AguaRESUMEN
Today, biofuels represent a hot topic in the context of petroleum and adjacent products decrease. As biofuels production increase, so does the production of their major byproduct, namely crude glycerol. The efficient usage of raw glycerol will concur to the biodiesel viability. As an inevitable waste of biodiesel manufacturing, glycerol is potentially an attractive substrate for the production of value-added products by fermentation processes, due to its large amounts, low cost and high degree of reduction. One of the most important usages of glycerol is its bioconversion through microbial fermentation to value-added materials like 1,3-propanediol and citric acid. There is a considerable industrial interest in 1,3-propanediol and citric acid production based on microbial fermentations, as it seems to be in competition with traditional technologies utilized for these products. In the present work, yields and concentrations of 1,3-propanediol and citric acid registered for different isolated strains are also described. Microbial bioconversion of glycerol represents a remarkable choice to add value to the biofuel production chain, allowing the biofuel industry to be more competitive. The current review presents certain ways for the bioconversion of crude glycerol into citric acid and 1,3-propanediol with high yields and concentrations achieved by using isolated microorganisms.
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Biocombustibles , Ácido Cítrico/metabolismo , Glicerol/metabolismo , Glicoles de Propileno/metabolismo , Bacterias/metabolismo , Biotecnología , Fermentación , Glicerol/química , Microbiología Industrial/métodos , Glicoles de Propileno/análisisRESUMEN
The applicability of 3D map modelling for melting point prediction was studied. The melting points in the ammonium polyphosphate-pentaerythritol-melamine chemical system of intumescent flame-retardant coatings over a wide range of concentrations were collected. The ternary diagram (triangle) of the melting points was plotted and an approximated 3D map was built for the range 205-345°C. The present work contains the thermal data for the observed ternary system and provides a new graphic system for making predictions for intumescent flame-retardant coatings. The applicability of the calculated 3D map for obtaining experimental samples of fire-retardant paints with a low melting point for thin steel constructions was shown.
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Retardadores de Llama/análisis , Polifosfatos/análisis , Glicoles de Propileno/análisis , Relación Estructura-Actividad Cuantitativa , Temperatura de Transición , Triazinas/análisis , Compuestos de AmonioRESUMEN
Coiled tubing corrosion was investigated for 16 field water samples (S5 to S20) from a Canadian shale gas field. Weight loss corrosion rates of carbon steel beads incubated with these field water samples averaged 0.2 mm/yr, but injection water sample S19 had 1.25±0.07 mm/yr. S19 had a most probable number of zero acid-producing bacteria and incubation of S19 with carbon steel beads or coupons did not lead to big changes in microbial community composition. In contrast other field water samples had most probable numbers of APB of 102/mL to 107/mL and incubation of these field water samples with carbon steel beads or coupons often gave large changes in microbial community composition. HPLC analysis indicated that all field water samples had elevated concentrations of bromide (average 1.6 mM), which may be derived from bronopol, which was used as a biocide. S19 had the highest bromide concentration (4.2 mM) and was the only water sample with a high concentration of active bronopol (13.8 mM, 2760 ppm). Corrosion rates increased linearly with bronopol concentration, as determined by weight loss of carbon steel beads, for experiments with S19, with filtered S19 and with bronopol dissolved in defined medium. This indicated that the high corrosion rate found for S19 was due to its high bronopol concentration. The corrosion rate of coiled tubing coupons also increased linearly with bronopol concentration as determined by electrochemical methods. Profilometry measurements also showed formation of multiple pits on the surface of coiled tubing coupon with an average pit depth of 60 µm after 1 week of incubation with 1 mM bronopol. At the recommended dosage of 100 ppm the corrosiveness of bronopol towards carbon steel beads was modest (0.011 mm/yr). Higher concentrations, resulting if biocide is added repeatedly as commonly done in shale gas operations, are more corrosive and should be avoided. Overdosing may be avoided by assaying the presence of residual biocide by HPLC, rather than by assaying the presence of residual surviving bacteria.
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Corrosión , Desinfectantes/análisis , Yacimiento de Petróleo y Gas/química , Acero , Microbiología del Agua , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bromuros/análisis , Canadá , Carbono/química , Cromatografía Líquida de Alta Presión , Código de Barras del ADN Taxonómico/métodos , Yacimiento de Petróleo y Gas/microbiología , Densidad de Población , Dinámica Poblacional , Glicoles de Propileno/análisis , ARN Ribosómico 16S/genética , Agua/análisisRESUMEN
Crude glycerol is an ideal feedstock for bioproduction of 1,3-propanediol (1,3-PDO) while pure culture always shows low substrate tolerance and limited productivity. In this study, an anaerobic microbial consortium for conversion of crude glycerol was selected and its 1,3-PDO production capacity was evaluated. The consortium was obtained from anaerobic activated sludge by 19 serial transfers and mainly consisted of 94.64% Clostridiaceae and 4.47% Peptostreptococcaceae. The consortium adapted well with high glycerol concentration of 120 g/L as well as wide substrate concentration fluctuation from 15 to 80 g/L, producing 60.61 and 82.66 g/L 1,3-PDO in the batch and fed-batch fermentation, with the productivity of 3.79 and 3.06 g/(Lâh), respectively, which are among the best results published so far. Furthermore, mini consortia isolated by serial dilution exhibited similar microbial composition but gradually decreasing tolerance to crude glycerol. Four randomly selected Clostridium butyricum displayed different substrate tolerance and insufficient 1,3-PDO production capacity. This work demonstrated that the high adaptation to crude glycerol of the consortium was the collaborative effort of different individuals.
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Adaptación Fisiológica , Glicerol/metabolismo , Consorcios Microbianos/fisiología , Glicoles de Propileno/metabolismo , Aclimatación , Anaerobiosis , Biocombustibles , Clostridiaceae/metabolismo , Clostridium butyricum/metabolismo , Medios de Cultivo , Fermentación , Glicerol/análisis , Peptostreptococcus/metabolismo , Glicoles de Propileno/análisisRESUMEN
Fermentation of wheat with yeast produces thin stillage (W-TS) and distiller's wet grains. A subsequent fermentation of W-TS (two-stage fermentation, TSF) with endemic bacteria at 25 and 37 °C decreased glycerol and lactic acid concentrations, while 1,3-propanediol (1,3-PD) and acetic acid accumulated with greater 1,3-PD and acetic acid produced at 37 °C. During TSF, W-TS colloids coagulated and floated in the fermentation medium producing separable liquid and slurry fractions. The predominant endemic bacteria in W-TS were Lactobacillus panis, L. gallinarum, and L. helveticus, and this makeup did not change substantially as fermentation progressed. As nutrients were exhausted, floating particles precipitated. Protein contents of slurry and clarified liquid increased and decreased, respectively, as TSF progressed. The liquid was easily filtered through an ultrafiltration membrane. These results suggested that TSF is a novel method for W-TS clarification and production of protein concentrates and 1,3-PD from W-TS.
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Lactobacillus/metabolismo , Proteínas de Plantas/metabolismo , Glicoles de Propileno/análisis , Triticum/metabolismo , Ácido Acético/metabolismo , Fermentación , Glicerol/metabolismo , Ácido Láctico/metabolismo , Lactobacillus/genética , Lactobacillus/aislamiento & purificación , Proteínas de Plantas/análisis , Glicoles de Propileno/metabolismo , Triticum/químicaRESUMEN
This paper addresses the problem of identifying the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae for production of 1,3-propanediol (1,3-PD) under different culture conditions. To achieve this goal, we propose a multi-level programming model. This model includes three optimization problems, where the constraint region of the first level problem is implicitly determined by the other two optimization problems. The optimized objectives of the first and second level problems are to minimize the set of fluxes that are of major importance to glycerol metabolism and the difference between the observed fluxes and those computed by the model, respectively. The third level problem in the proposed multi-level programming simultaneously solves a set of flux balance analysis (FBA) models. A method is proposed to solve efficiently the presented multi-level programming problem. In this method, we first transform the proposed multi-level problem into a bi-level problem by applying the dual theory of linear programming to the FBA models of the third level. Next, the optimal solution of the above bi-level problem is obtained by iteratively solving a sequence of mixed integer programming problems. Optimization results reveal that the proposed method can identify the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae under three groups of experimental data.
Asunto(s)
Glicerol/metabolismo , Klebsiella pneumoniae/metabolismo , Modelos Biológicos , Glicoles de Propileno/metabolismo , Algoritmos , Biología Computacional , Análisis de Flujos Metabólicos , Redes y Vías Metabólicas , Glicoles de Propileno/análisisRESUMEN
The aim of this study has been to produce 1,3-propanediol (1,3-PDO) from glycerol (gly) fermentation by means of a microbial mixed culture (granular sludge), as well as to establish the operational conditions of two up-flow anaerobic sludge blanket (UASB) reactors in order to achieve a maximum 1,3-PDO yield. The UASB reactors with initial pH values set at 6.8 and 5.5 were operated at 30 °C during 165 days. Thirteen variables were previously screened by a Plackett-Burman (PB) design; results showed that yeast extract, MgSO4 and methanogenesis inhibition (by heat shock) showed a positive effect, whereas high glycerol concentration, tryptone and CaCl2 showed a negative impact on the 1,3-PDO produced by glycerol degradation. Following four experimental periods, the highest average yield of 0.43 mol 1,3-PDO mol-1 gly was achieved when sodium bicarbonate was added to the reactors. Propionate and acetate were also produced and a high microorganism diversity was detected; however, the restrictive operational conditions of the reactors led to the death of the methanogenic archaea. Nevertheless, the continuous production of 1,3-PDO from glycerol within UASB reactors inoculated with granular sludge can be considered highly feasible.