scholarly journals Genome-Based Metabolic Engineering of Mannheimia succiniciproducens for Succinic Acid Production

2006 ◽  
Vol 72 (3) ◽  
pp. 1939-1948 ◽  
Author(s):  
Sang Jun Lee ◽  
Hyohak Song ◽  
Sang Yup Lee
Keyword(s):  
Succinic Acid ◽  
Acid Production ◽  
Gene Knockout ◽  
Product Formation ◽  
Anaerobic Growth ◽  
Fermentation Products ◽  
Acid Yield ◽  
Mannheimia Succiniciproducens ◽  
Succinic Acid Production ◽  
Pep Carboxykinase

ABSTRACT Succinic acid is a four-carbon dicarboxylic acid produced as one of the fermentation products of anaerobic metabolism. Based on the complete genome sequence of a capnophilic succinic acid-producing rumen bacterium, Mannheimia succiniciproducens, gene knockout studies were carried out to understand its anaerobic fermentative metabolism and consequently to develop a metabolically engineered strain capable of producing succinic acid without by-product formation. Among three different CO2-fixing metabolic reactions catalyzed by phosphoenolpyruvate (PEP) carboxykinase, PEP carboxylase, and malic enzyme, PEP carboxykinase was the most important for the anaerobic growth of M. succiniciproducens and succinic acid production. Oxaloacetate formed by carboxylation of PEP was found to be converted to succinic acid by three sequential reactions catalyzed by malate dehydrogenase, fumarase, and fumarate reductase. Major metabolic pathways leading to by-product formation were successfully removed by disrupting the ldhA, pflB, pta, and ackA genes. This metabolically engineered LPK7 strain was able to produce 13.4 g/liter of succinic acid from 20 g/liter glucose with little or no formation of acetic, formic, and lactic acids, resulting in a succinic acid yield of 0.97 mol succinic acid per mol glucose. Fed-batch culture of M. succiniciproducens LPK7 with intermittent glucose feeding allowed the production of 52.4 g/liter of succinic acid, with a succinic acid yield of 1.16 mol succinic acid per mol glucose and a succinic acid productivity of 1.8 g/liter/h, which should be useful for industrial production of succinic acid.

scholarly journals Metabolic Engineering of Escherichia coli for Enhanced Production of Succinic Acid, Based on Genome Comparison and In Silico Gene Knockout Simulation

2005 ◽  
Vol 71 (12) ◽  
pp. 7880-7887 ◽  
Author(s):  
Sang Jun Lee ◽  
Dong-Yup Lee ◽  
Tae Yong Kim ◽  
Byung Hun Kim ◽  
Jinwon Lee ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Succinic Acid ◽  
In Silico ◽  
Acid Production ◽  
Gene Knockout ◽  
Fermentation Products ◽  
E Coli ◽  
Metabolic Analysis ◽  
Enhanced Production ◽  
Succinic Acid Production

ABSTRACT Comparative analysis of the genomes of mixed-acid-fermenting Escherichia coli and succinic acid-overproducing Mannheimia succiniciproducens was carried out to identify candidate genes to be manipulated for overproducing succinic acid in E. coli. This resulted in the identification of five genes or operons, including ptsG, pykF, sdhA, mqo, and aceBA, which may drive metabolic fluxes away from succinic acid formation in the central metabolic pathway of E. coli. However, combinatorial disruption of these rationally selected genes did not allow enhanced succinic acid production in E. coli. Therefore, in silico metabolic analysis based on linear programming was carried out to evaluate the correlation between the maximum biomass and succinic acid production for various combinatorial knockout strains. This in silico analysis predicted that disrupting the genes for three pyruvate forming enzymes, ptsG, pykF, and pykA, allows enhanced succinic acid production. Indeed, this triple mutation increased the succinic acid production by more than sevenfold and the ratio of succinic acid to fermentation products by ninefold. It could be concluded that reducing the metabolic flux to pyruvate is crucial to achieve efficient succinic acid production in E. coli. These results suggest that the comparative genome analysis combined with in silico metabolic analysis can be an efficient way of developing strategies for strain improvement.

From genome sequence to integrated bioprocess for succinic acid production by Mannheimia succiniciproducens

2008 ◽  
Vol 79 (1) ◽  
pp. 11-22 ◽  
Author(s):  
Sang Yup Lee ◽  
Ji Mahn Kim ◽  
Hyohak Song ◽  
Jeong Wook Lee ◽  
Tae Yong Kim ◽  
...  
Keyword(s):  
Succinic Acid ◽  
Genome Sequence ◽  
Acid Production ◽  
Mannheimia Succiniciproducens ◽  
Succinic Acid Production ◽  
Integrated Bioprocess

scholarly journals Influence of the Initial Sugar Concentration and Supplementation with Yeast Extract on the Succinic Acid Fermentation from Lactose Based Medium

2021 ◽  
Author(s):  
Christiane Terboven ◽  
Christian Abendroth ◽  
Janin Laumer ◽  
Christiane Herrmann ◽  
Roland Schneider ◽  
...  
Keyword(s):  
Succinic Acid ◽  
Yeast Extract ◽  
Acid Production ◽  
Sugar Concentration ◽  
Actinobacillus Succinogenes ◽  
Sugar Consumption ◽  
Acid Fermentation ◽  
Acid Yield ◽  
Succinic Acid Production ◽  
Succinic Acid Fermentation

The aim of this study was to investigate succinic acid production from lactose concentrate, a by-product of cheese-making, using Actinobacillus succinogenes and Basfia succiniciproducens. Although the capability of these strains to metabolize different sugars is already known, their application in the conversion of lactose bears high potential for optimization. With regard to B. succiniciproducens this approach is completely novel. In particular the influence of the mediums sugar concentration and its supplementation with yeast extract to prevent a lack of proteins and vitamins were examined. Lactose based media containing sugar concentrations between 20 and 65 g L-1 and 5 g L-1 yeast extract were fermented, whereby both strains showed comparable performances. The best results in succinic acid yield and acid concentration, 0.57 g g-1 initial sugar and 23 g L1, were achieved at an initial sugar concentration of 43 g L-1. The necessity of yeast extract was demonstrated using the sugar optimized medium without supplementation. As a result, yield and concentration of succinic acid dropped to 0.34 g g-1 and 13 g L-1, the sugar consumption decreased from more than 99 to less than 55 %. Therefore the supplementation amount of 5 g L-1 yeast extract can be regarded as well-balanced.

scholarly journals Improved Succinic Acid Production in the Anaerobic Culture of an Escherichia coli pflB ldhA Double Mutant as a Result of Enhanced Anaplerotic Activities in the Preceding Aerobic Culture

2007 ◽  
Vol 73 (24) ◽  
pp. 7837-7843 ◽  
Author(s):  
Hui Wu ◽  
Zhi-min Li ◽  
Li Zhou ◽  
Qin Ye
Keyword(s):  
Escherichia Coli ◽  
Succinic Acid ◽  
Acid Production ◽  
Double Mutant ◽  
Anaerobic Culture ◽  
Two Stage ◽  
Acid Yield ◽  
Succinic Acid Production ◽  
Ferment Glucose ◽  
Two Stage Culture

ABSTRACT Escherichia coli NZN111 is a pflB ldhA double mutant which loses its ability to ferment glucose anaerobically due to redox imbalance. In this study, two-stage culture of NZN111 was carried out for succinic acid production. It was found that when NZN111 was aerobically cultured on acetate, it regained the ability to ferment glucose with succinic acid as the major product in subsequent anaerobic culture. In two-stage culture carried out in flasks, succinic acid was produced at a level of 11.26 g/liter from 13.4 g/liter of glucose with a succinic acid yield of 1.28 mol/mol glucose and a productivity of 1.13 g/liter·h in the anaerobic stage. Analyses of key enzyme activities revealed that the activities of isocitrate lyase, malate dehydrogenase, malic enzyme, and phosphoenolpyruvate (PEP) carboxykinase were greatly enhanced while those of pyruvate kinase and PEP carboxylase were reduced in the acetate-grown cells. The two-stage culture was also performed in a 5-liter fermentor without separating the acetate-grown NZN111 cells from spent medium. The overall yield and concentration of succinic acid reached 1.13 mol/mol glucose and 28.2 g/liter, respectively, but the productivity of succinic acid in the anaerobic stage dropped to 0.7 g/liter·h due to cell autolysis and reduced anaplerotic activities. The results indicate the great potential to take advantage of cellular regulation mechanisms for improvement of succinic acid production by a metabolically engineered E. coli strain.

Homo-succinic acid production by metabolically engineered Mannheimia succiniciproducens

2016 ◽  
Vol 38 ◽  
pp. 409-417 ◽  
Author(s):  
Jeong Wook Lee ◽  
Jongho Yi ◽  
Tae Yong Kim ◽  
Sol Choi ◽  
Jung Ho Ahn ◽  
...  
Keyword(s):  
Succinic Acid ◽  
Acid Production ◽  
Mannheimia Succiniciproducens ◽  
Succinic Acid Production
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