Metabolic Engineering of
            <i>Escherichia coli</i>
            for Efficient Conversion of Glycerol to Ethanol

Trinh, Cong T.; Srienc, Friedrich

doi:10.1128/aem.00670-09

Cited by 135 publications

(95 citation statements)

References 29 publications

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“…Anoxic fermentation of glycerol by E. coli has received much attention in the last few years, and its use in industrial bioprocesses was analysed Gonzalez 2007, 2008;Gonzalez et al 2008;Murarka et al 2008;Trinh and Srienc 2009). Most of the results published regarding metabolic engineering of E. coli for ethanol synthesis describe experiments conducted under strict anoxic conditions.…”

Section: Discussionmentioning

confidence: 99%

Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides

Nikel

Ramírez

Pettinari

et al. 2010

Journal of Applied Microbiology

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Aims: Analysis of the physiology and metabolism of Escherichia coli arcA and creC mutants expressing a bifunctional alcohol‐acetaldehyde dehydrogenase from Leuconostoc mesenteroides growing on glycerol under oxygen‐restricted conditions. The effect of an ldhA mutation and different growth medium modifications was also assessed. Methods and Results: Expression of adhE in E. coli CT1061 [arcA creC(Con)] resulted in a 1·4‐fold enhancement in ethanol synthesis. Significant amounts of lactate were produced during micro‐oxic cultures and strain CT1061LE, in which fermentative lactate dehydrogenase was deleted, produced up to 6·5 ± 0·3 g l−1 ethanol in 48 h. Escherichia coli CT1061LE derivatives resistant to >25 g l−1 ethanol were obtained by metabolic evolution. Pyruvate and acetaldehyde addition significantly increased both biomass and ethanol concentrations, probably by overcoming acetyl‐coenzyme A (CoA) shortage. Yeast extract also promoted growth and ethanol synthesis, and this positive effect was mainly attributable to its vitamin content. Two‐stage bioreactor cultures were conducted in a minimal medium containing 100 μg l−1 calcium d‐pantothenate to evaluate oxic acetyl‐CoA synthesis followed by a switch into fermentative conditions. Ethanol reached 15·4 ± 0·9 g l−1 with a volumetric productivity of 0·34 ± 0·02 g l−1 h−1. Conclusions: Escherichia coli responded to adhE over‐expression by funnelling carbon and reducing equivalents into a highly reduced metabolite, ethanol. Acetyl‐CoA played a key role in micro‐oxic ethanol synthesis and growth. Significance and Impact of the Study: Insight into the micro‐oxic metabolism of E. coli growing on glycerol is essential for the development of efficient industrial processes for reduced biochemicals production from this substrate, with special relevance to biofuels synthesis.

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Section: Discussionmentioning

confidence: 99%

Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides

Nikel

Ramírez

Pettinari

et al. 2010

Journal of Applied Microbiology

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“…pneumoniae CLG29 was isolated from bryophytes grown on the base of leaf stalks of Terminalia catappa at UNESP -Universidade Estadual Paulista, Rio Claro, Brazil and characterized as a new 1,3-PDO producer (da Silva et al, 2014). E. coli TCS099 -ΔmgsA, ΔldhA, ΔfdrA, Δzwf, Δndh, ΔmaeB, Δpta, ΔpoxB, ΔmhpF, ΔadhP and ΔadhE (Trinh and Srienc, 2009), was used at University of Tennessee Knoxville, and E. coli SZ63, W3110 mutant, ΔfocA-pflB::FRT, ΔfrdBC ΔadhE::FRT, ackA::FRT (Zhou et al, 2003) was kindly sent from the University of Florida to the Department of Biochemistry and Microbiology, Biosciences Institute of Rio Claro, Univ. Estadual Paulista -UNESP.…”

Section: Strains and Maintenancementioning

confidence: 99%

Anaerobic and micro-aerobic 1,3-propanediol production by engineered Escherichia coli with dha genes from Klebsiella pneumoniae GLC29

Neto¹,

Briganti²,

Layton³

et al. 2017

Afr. J. Biotechnol.

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1,3-Propanediol (1,3-PDO) is a bifunctional molecule, and used in applications similar to those of ethylene glycol, propylene glycol, 1,3-butanediol and 1,4-butanediol. The use of glycerol as a feedstock is an alternative to reduce production costs for both 1,3-PDO and biodiesel, since biodiesel glycerol can be used for the production of 1,3-PDO by bacteria. Also, using metabolic engineering, it is possible to manipulate the metabolic routes and obtain high value products, reduce or eliminate the formation of undesirable byproducts. The aim of the study was to produce 1,3-propanediol in E. coli cloned with dha genes from Klebsiella pneumoniae GLC29. Six genes responsible for 1,3-PDO production in Klebsiella pneumoniae GLC29 were cloned. These genes were assembled in pSB1C3 as an expression vector: Genes dhaB1, dhaB2, dhaB3 and dhaT (pSB1C3dhaB123T), and another vector with genes dhaF and dhaG (pSB1C3dhaB123TFG) were derived using Gibson's Assembly technique. Escherichia coli TCS099 and SZ63 stains were used as hosts for 1,3-PDO production, and kept at -80°C for long-term storage. Glycerol was used as the sole or main carbon source in all experiments. Fermentations were performed in flasks in aerobic and anaerobic conditions using minimal media. Also, two stage fermentation (aerobic-anaerobic) was performed for 1,3-propanediol production. Only pSB1C3dhaB123TFG was able to produce high amounts of 1,3-PDO in shake flasks experiments, producing 2.5 g/L in micro-aerobic conditions, using E. coli TCS099 as host. Besides, E. coli SZ63 hosting pSB1C3dhaB123TFG was able to produce high amounts of 1,3-PDO, corresponding to 11.3 g/L of 1,3-PDO using a two-stage fermentation process using low concentration of vitamin B12 (1 mg/L). Plasmid pSB1C3dhaB123TFG shows potential for producing high amounts of 1,3-PDO, specially because of dhaF and dhaG, reaffirming the importance of this genes on 1,3-PDO production, especially with the addition of low amounts of vitamin B12, which is an expensive compound.

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“…Other applications include biofuels, such as bioethanol (156) or biohydrogen (157), and drug target discovery (158,159). Recent reviews discussing the various applications of GSMMs are available (20,48,(160)(161)(162).…”

Section: Applications and Discussionmentioning

confidence: 99%

In SilicoConstraint-Based Strain Optimization Methods: the Quest for Optimal Cell Factories

Maia

Rocha

2016

Microbiol Mol Biol Rev

116

107

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SUMMARYShifting from chemical to biotechnological processes is one of the cornerstones of 21st century industry. The production of a great range of chemicals via biotechnological means is a key challenge on the way toward a bio-based economy. However, this shift is occurring at a pace slower than initially expected. The development of efficient cell factories that allow for competitive production yields is of paramount importance for this leap to happen. Constraint-based models of metabolism, together within silicostrain design algorithms, promise to reveal insights into the best genetic design strategies, a step further toward achieving that goal. In this work, a thorough analysis of the mainin silicoconstraint-based strain design strategies and algorithms is presented, their application in real-world case studies is analyzed, and a path for the future is discussed.

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Metabolic Engineering of Escherichia coli for Efficient Conversion of Glycerol to Ethanol

Cited by 135 publications

References 29 publications

Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides

Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides

Anaerobic and micro-aerobic 1,3-propanediol production by engineered Escherichia coli with dha genes from Klebsiella pneumoniae GLC29

In SilicoConstraint-Based Strain Optimization Methods: the Quest for Optimal Cell Factories

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