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Scale-up of anaerobic 1,3-propanediol production by Clostridium butyricum DSP1 from crude glycerol.

ABSTRACT: BACKGROUND: As the production of biofuels from raw materials continuously increases, optimization of production processes is necessary. A very important issue is the development of wasteless methods of biodiesel production. One way of utilization of glycerol generated in biodiesel production is its microbial conversion to 1,3-PD (1,3-propanediol). RESULTS: The study investigated the scale-up of 1,3-PD synthesis from crude glycerol by Clostridium butyricum. Batch fermentations were carried out in 6.6 L, 42 L and 150 L bioreactors. It was observed that cultivation of C. butyricum on a pilot scale did not decrease the efficiency of 1,3-PD production. The highest concentrations of 1,3-PD, 37 g/L for batch fermentation and 71 g/L for fed-batch fermentation, were obtained in the 6.6 L bioreactor. The kinetic parameters of 1,3-PD synthesis from crude glycerol established for batch fermentation were similar regarding all three bioreactor capacities. During fed-batch fermentation, the concentration of 1,3-PD in the 150 L bioreactor was lower and the substrate was not completely utilized. That suggested the presence of multifunctional environmental stresses in the 150 L bioreactor, which was confirmed by protein analysis. CONCLUSION: The values of effectivity parameters for 1,3-PD synthesis in batch fermentations carried out in 6.6 L, 42 L and 150 L bioreactors were similar. The parameters obtained during fed-batch fermentations in the 150 L bioreactor differed in the rate and percentage of substrate utilization. The analysis of cell proteins demonstrated that a number of multifunctional stresses occurred during fed-batch fermentations in the 150 L bioreactor, which suggests the possibility of identifying the key stages in the biochemical process where inhibition of 1,3-PD synthesis pathways can be observed.

SUBMITTER: Szymanowska-Powalowska D

PROVIDER: S-EPMC3974118 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Scale-up of anaerobic 1,3-propanediol production by Clostridium butyricum DSP1 from crude glycerol.

Szymanowska-Powałowska Daria D Białas Wojciech W

BMC microbiology 20140220

<h4>Background</h4>As the production of biofuels from raw materials continuously increases, optimization of production processes is necessary. A very important issue is the development of wasteless methods of biodiesel production. One way of utilization of glycerol generated in biodiesel production is its microbial conversion to 1,3-PD (1,3-propanediol).<h4>Results</h4>The study investigated the scale-up of 1,3-PD synthesis from crude glycerol by Clostridium butyricum. Batch fermentations were c ...[more]

PMID: 24555775

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Project description:BackgroundOscillation is a special cell behavior in microorganisms during continuous fermentation, which poses threats to the output stability for industrial productions of biofuels and biochemicals. In previous study, a spontaneous oscillatory behavior was observed in Clostridium butyricum-intensive microbial consortium in continuous fermentation for 1,3-propanediol (1,3-PDO) production from glycerol, which led to the discovery of oscillation in species C. butyricum.ResultsSpontaneous oscillations by C. butyricum tended to occur under glycerol-limited conditions at low dilution rates. At a glycerol feed concentration of 88 g/L and a dilution rate of 0.048 h-1, the oscillatory behavior of C. butyricum was observed after continuous operation for 146 h and was sustained for over 450 h with an average oscillation period of 51 h. During oscillations, microbial glycerol metabolism exhibited dramatic periodic changes, in which productions of lactate, formate and hydrogen significantly lagged behind that of other products including biomass, 1,3-PDO and butyrate. Analysis of extracellular oxidation-reduction potential and intracellular ratio of NAD+/NADH indicated that microbial cells experienced distinct redox changes during oscillations, from oxidized to reduced state with decreasing of growth rate. Meanwhile, C. butyricum S3 exhibited periodic morphological changes during oscillations, with aggregates, elongated shape, spores or cell debris at the trough of biomass production. Transcriptome analysis indicated that expression levels of multiple genes were up-regulated when microbial cells were undergoing stress, including that for pyruvate metabolism, conversion of acetyl-CoA to acetaldehyde as well as stress response.ConclusionThis study for the first time systematically investigated the oscillatory behavior of C. butyricum in aspect of occurrence condition, metabolism, morphology and transcriptome. Based on the experimental results, two hypotheses were put forward to explain the oscillatory behavior: disorder of pyruvate metabolism, and excessive accumulation of acetaldehyde.

Project description:Crude glycerol is largely generated as the main by-product of the biodiesel industry and is unprofitable for industrial application without costly purification. The direct bioconversion of crude glycerol into 1,3-propanediol (1,3-PDO) by microorganisms is a promising alternative for effective and economic utilization. In this study, Klebsiella pneumoniae 2e was newly isolated for the conversion of crude glycerol into 1,3-PDO. Batch fermentation analysis confirmed that crude glycerol and its main impurities had slight impacts on the growth, key enzyme activity, and 1,3-PDO production of K. pneumoniae 2e. The 1,3-PDO yield from crude glycerol by K. pneumoniae 2e reached 0.64 mol 1,3-PDO/mol glycerol, which was higher than that by most reported 1,3-PDO-producing Klebsiella strains. Genomic profiling revealed that K. pneumoniae 2e possesses 30 genes involved in glycerol anaerobic metabolism and 1,3-PDO biosynthesis. Quantitative real-time PCR analysis of these genes showed that the majority of the genes encoding the key enzymes for glycerol metabolism and 1,3-PDO biosynthesis were significantly upregulated during culture in crude glycerol relative to that in pure glycerol. Further comparative genomic analysis revealed a novel glycerol uptake facilitator protein in K. pneumoniae 2e and a higher number of stress response proteins than in other Klebsiella strains. This work confirms the adaptability of a newly isolated 1,3-PDO-producing strain, K. pneumoniae 2e, to crude glycerol and provides insights into the molecular mechanisms involved in its crude glycerol tolerance, which is valuable for industrial 1,3-PDO production from crude glycerol.IMPORTANCE The rapid development of the biodiesel industry has led to tremendous crude glycerol generation. Due to the presence of complex impurities, crude glycerol has low value for industry without costly purification. Obtaining novel microorganisms capable of direct and efficient bioconversion of crude glycerol to value-added products has great economic potential for industrial application. In this work, we characterized a newly isolated strain, Klebsiella pneumoniae 2e, with the capacity to efficiently produce 1,3-propanediol (1,3-PDO) from crude glycerol and demonstrated its adaptation to crude glycerol. Our work provides insights into the molecular mechanisms of K. pneumoniae 2e adaptation to crude glycerol and the expression patterns of its genes involved in 1,3-PDO biosynthesis, which will contribute to the development of industrial 1,3-PDO production from crude glycerol.

Dataset Information

Scale-up of anaerobic 1,3-propanediol production by Clostridium butyricum DSP1 from crude glycerol.

Publications

Scale-up of anaerobic 1,3-propanediol production by Clostridium butyricum DSP1 from crude glycerol.

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