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1.
Bioprocess Biosyst Eng ; 41(12): 1793-1805, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30173374

RESUMEN

To investigate the relationship between the yield of 1,3-propanediol (1,3-PD) and the flux variation in metabolic pathways of Klebsiella pneumoniae, an optimized calculation method was constructed on basis of dynamic flux balance analysis by combining genome-scale flux balance analysis with a kinetic model of extracellular metabolites. Through optimizing calculations, a more completely expanded metabolic pathway was obtained, which includes the previously reported metabolic pathway and additional three pathways or site: a pentose phosphate pathway (PPP) elicited at the dihydroxyacetone (DHA) node to provide more reducing equivalents; a branch of synthetic amino acids at the 3-phosphoglycerate (3PG) node; and the α-ketoglutarate site in the tricarboxylic acid (TCA) cycle leading to anabolic pathways for glutamate and other amino acids. On this basis, the relationships between the dynamic flux distribution of the important nodes in the metabolic pathway and the yield of 1,3-propanediol were analyzed. First, dynamic flux change from DHA to the PPP is positively correlated with the yield. Second, variation in flux in the TCA cycle is also positively correlated with the yield of 1,3-propanediol. In addition, the influence of the feedback loop formed by the cofactor tetrahydrofolate on the flux change of TCA in the amino acid anabolic pathway was examined. These results are of important reference value and have guiding significance for the extension of the glycerol metabolism pathway in K. pneumoniae, the rational transformation of genetic engineering in bacteria, and the optimization of metabolic pathways for industrial production.


Asunto(s)
Ciclo del Ácido Cítrico , Glicerol/metabolismo , Klebsiella pneumoniae/metabolismo , Vía de Pentosa Fosfato , Glicoles de Propileno/metabolismo
2.
Biotechnol Biofuels Bioprod ; 17(1): 38, 2024 Mar 07.
Artículo en Inglés | MEDLINE | ID: mdl-38454489

RESUMEN

BACKGROUND: Glycerol, as a by-product, mainly derives from the conversion of many crops to biodiesel, ethanol, and fatty ester. Its bioconversion to 1,3-propanediol (1,3-PDO) is an environmentally friendly method. Continuous fermentation has many striking merits over fed-batch and batch fermentation, such as high product concentration with easy feeding operation, long-term high productivity without frequent seed culture, and energy-intensive sterilization. However, it is usually difficult to harvest high product concentrations. RESULTS: In this study, a three-stage continuous fermentation was firstly designed to produce 1,3-PDO from crude glycerol by Clostridium butyricum, in which the first stage fermentation was responsible for providing the excellent cells in a robust growth state, the second stage focused on promoting 1,3-PDO production, and the third stage aimed to further boost the 1,3-PDO concentration and reduce the residual glycerol concentration as much as possible. Through the three-stage continuous fermentation, 80.05 g/L 1,3-PDO as the maximum concentration was produced while maintaining residual glycerol of 5.87 g/L, achieving a yield of 0.48 g/g and a productivity of 3.67 g/(L·h). Based on the 14 sets of experimental data from the first stage, a kinetic model was developed to describe the intricate relationships among the concentrations of 1,3-PDO, substrate, biomass, and butyrate. Subsequently, this kinetic model was used to optimize and predict the highest 1,3-PDO productivity of 11.26 g/(L·h) in the first stage fermentation, while the glycerol feeding concentration and dilution rate were determined to be 92 g/L and 0.341 h-1, separately. Additionally, to achieve a target 1,3-PDO production of 80 g/L without the third stage fermentation, the predicted minimum volume ratio of the second fermenter to the first one was 11.9. The kinetics-based two-stage continuous fermentation was experimentally verified well with the predicted results. CONCLUSION: A novel three-stage continuous fermentation and a kinetic model were reported. Then a simpler two-stage continuous fermentation was developed based on the optimization of the kinetic model. This kinetics-based development of two-stage continuous fermentation could achieve high-level production of 1,3-PDO. Meanwhile, it provides a reference for other bio-chemicals production by applying kinetics to optimize multi-stage continuous fermentation.

3.
Biotechnol Prog ; 40(1): e3411, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-37985220

RESUMEN

To study the relationship between the yield of 1,3-propanediol (1,3-PDO) and the flux change of the Clostridium butyricum metabolic pathway, an optimized calculation method based on dynamic flux balance analysis was used by combining genome-scale flux balance analysis with a kinetic model. A more comprehensive and extensive metabolic pathway was obtained by optimization calculations. The primary extended branches include: the dihydroxyacetone node, which enters the pentose phosphate pathway; the α-oxoglutarate node, which has synthetic metabolic pathways for glutamic acid and amino acids; and the serine and homocysteine nodes, which produce cystathionine before homocysteine enters the methionine cycle pathway. According to the expanded metabolic network, the flux distribution of key nodes in the metabolic pathway and the relationship between the flux distribution ratio of nodes and the yield of 1,3-PDO were analyzed. At the dihydroxyacetone node, the flux of dihydroxyacetone converted to dihydroxyacetone phosphate was positively correlated with the yield of 1,3-PDO. As an important intermediate product, the flux change in the metabolic pathway of α-oxoglutarate reacting with amino acids to produce glutamic acid is positively correlated with the yield. When pyruvate was used as the central node to convert into lactic acid and α-oxoglutarate, the proportion of branch flux was negatively correlated with the yield of 1,3-PDO. These studies provide a theoretical basis for the optimization and further study of the metabolic pathway of C. butyricum.


Asunto(s)
Clostridium butyricum , Clostridium butyricum/metabolismo , Fermentación , Dihidroxiacetona , Ácidos Cetoglutáricos/metabolismo , Glicerol/metabolismo , Glicoles de Propileno , Propilenglicol/metabolismo , Homocisteína/metabolismo , Glutamatos/metabolismo
4.
Biotechnol Prog ; 38(1): e3225, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34775686

RESUMEN

In utilizing glycerol to produce 1,3-propanediol by microbial fermentation, the problems of low utilization rate and poor production performance need to be addressed. Based on the analysis of a mathematical model for 1,3-propanediol production from glycerol by Klebsiella pneumoniae, this study theoretically investigated the effects of the dilution rate and the initial glycerol concentration in a two-stage fermentation process and the feasibility of applying the feedback control methods. First, the optimal operation conditions of initial glycerol concentration and dilution rate were obtained. Through the use of feedback control theory, a control strategy for dilution rate was designed and optimized to shorten the settling time (time required for fermentation to reach stability) from 60.92 to 36.68 h for the first reactor, and from 53.66 to 22.68 h for the second reactor. In addition, the yield of 1,3-propanediol in both two reactors reached up to 0.5 g·g-1 . The simulation results indicated that the feedback control strategy for dilution rate increased the product concentration, reduced the residual glycerol in the fermentation broth, and greatly improved the performance of the fermentation. A feeding strategy of automatic control for dilution rate has been established and will be applied as an effective guiding scheme in automatic continuous fermentations for production of 1,3-propanediol.


Asunto(s)
Glicerol , Glicoles de Propileno , Retroalimentación , Fermentación , Klebsiella pneumoniae , Modelos Teóricos
5.
J Biotechnol ; 301: 68-78, 2019 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-31175893

RESUMEN

Using mathematical model and computer simulation to predict biological processes and optimize the target production is an important strategy for optimizing fermentation process. However, the inherent uncertainty of the kinetic model severely limits the predictive capability. In this study, optimize target production, such as productivity and yield of 1, 3-propanediol produced by Klebsiella pneumoniae using glycerol as substrate, the ensemble modeling approach was used to reduce the model's uncertainty for fermentation process as much as possible, and effectively improve its prediction performance. Firstly, through sensitivity analysis, the parameters having significant influence on the model were determined as the adjustable parameters for the ensemble modeling. After comparison, the appropriate threshold coefficient of the model error was determined, and the sampling method was used to generate as many equivalent parameter sets as possible. Each set of parameters was separately applied for the simulation, and all the predicted values were integrated for the weighted average. Therefore, the expected value of the prediction was obtained. Compared with the traditional simulation using single parameter set, the ensemble modeling method achieved the lower relative error between the prediction and the experimental value and the greatly improved model prediction performance. Moreover, the optimal productivity and yield of 1, 3-propanediol and the corresponding operating conditions were obtained, respectively. The ensemble modeling approach effectively compensates for the uncertainties of the model, making its prediction performance more practical, which is important for computer simulations to predict and guide the actual production process.


Asunto(s)
Glicerol/metabolismo , Klebsiella pneumoniae/metabolismo , Glicoles de Propileno/metabolismo , Biotecnología , Simulación por Computador , Fermentación , Modelos Estadísticos
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