Involvement of the AtoSCDAEB regulon in the high molecular weight poly-(R)-3-hydroxybutyrate biosynthesis in phaCAB+ Escherichia coli

Theodorou, Evaggelos C.; Θεοδώρου, Μάμας; Kyriakidis, Dimitrios A.

doi:10.1016/j.ymben.2012.03.010

Cited by 11 publications

(8 citation statements)

References 57 publications

(98 reference statements)

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“…Recombinant E. coli has been used for PHB production and PHB flux studies (Fidler and Dennis, 1992;Gao et al, 2011;Lee and Choi, 2001;Theodorou et al, 2012;Tyo et al, 2010). Effects of dissolved oxygen concentration on the growth of E. coli have been discussed (van Wegen et al, 2001;Wang and Lee, 1997).…”

Section: Introductionmentioning

confidence: 99%

Enhanced co-production of hydrogen and poly-(R)-3-hydroxybutyrate by recombinant PHB producing E. coli over-expressing hydrogenase 3 and acetyl-CoA synthetase

Wang

Shi

Chen

et al. 2012

Metabolic Engineering

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Section: Introductionmentioning

confidence: 99%

Enhanced co-production of hydrogen and poly-(R)-3-hydroxybutyrate by recombinant PHB producing E. coli over-expressing hydrogenase 3 and acetyl-CoA synthetase

Wang

Shi

Chen

et al. 2012

Metabolic Engineering

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“…There is typically an inherent tradeoff between overall productivity and substrate utilization in producing microbial PHB 29 , and it is possible to achieve high PHB productivities at the expense of substrate utilization [30][31][32] . However there remain application scenarios where a high substrate utilization and yield must be simultaneously maintained; in particular when substrate costs are high and in remote and austere environments such as long term missions in space where access to feedstock material might be limited.…”

Section: Discussionmentioning

confidence: 99%

“…To date, no engineering or interrogation of two component systems has been done in species of Cupriavidus for the purposes of increasing PHB or deciphering any TCS-related aspects of the native regulation of PHB synthesis/mobilization. However, overexpression of the native E. coli AtoSC two component system activating fatty acid biosynthesis genes in an E. coli mutant expressing C. necator phaCAB genes resulted in increased overall PHB titer and per-cell PHB accumulation 30 , linking native fatty acid metabolism to heterologous PHB production in the organism. More generally, TCS system engineering has resulted in bioproduction benefits for other products in other organisms as well such as the elimination of gacS - gacA system in P. putida resulting in an increased conversion of para-coumarate to indigoidine 46 .…”

Section: Discussionmentioning

confidence: 99%

Eliminating Genes for a Two Component System Increases PHB Productivity in Cupriavidus basilensis 4G11 Under PHB Suppressing, Non-Stress Conditions

Sander

Abel

Friedline

et al. 2022

Preprint

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Species of bacteria from the genus Cupriavidus are known, in part, for their ability to produce high amounts of poly-hydroxybutyrate (PHB) making them attractive candidate bioplastic producers. The native production of PHB occurs during periods of metabolic stress, and the process regulating the initiation of PHB accumulation in these organisms is not fully understood. Screening an RB-TnSeq transposon library of Cupriavidus basilensis 4G11 allowed us to identify two genes of an apparent, uncharacterized two component system which, when omitted from the genome, are capable of increased PHB productivity in balanced, non-stress growth conditions. We observe average increases in PHB productivity of 56% and 41% relative to the wildtype parent strain, upon deleting each of two genes individually from the genome. The increased PHB phenotype disappears, however, in nitrogen-free unbalanced growth conditions suggesting the phenotype is specific to fast-growing, replete, non-stress growth. Bioproduction modeling suggests this phenotype could be due to a decreased reliance on metabolic stress induced by nitrogen limitation to initiate PHB production in the mutant strains. Such strains may allow for the use of single stage, continuous bioreactor systems, which are far simpler than PHB bioproduction schemes used previously. Bioproductivity modeling suggests that omitting this regulation in the cells may increase PHB productivity up to 24% relative to the wildtype organism when using single stage continuous systems. This work furthermore expands our understanding of the regulation of PHB accumulation in Cupriavidus, in particular the initiation of this process upon transition into unbalanced growth regimes.

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“…Theodorou et al . [79] recently demonstrated that PHB synthesis from D-glucose is impaired in a Δ atoSC and Δ atoDAEB mutant, and that this phenotype can be restored by ectopic expression of Ato components in the recombinants. The authors hinted an important role of the AtoSC system in polymer accumulation and also proposed that this regulatory system could be an important target for metabolic engineering manipulations.…”

Section: Bacterial Polyhydroxyalkanoatesmentioning

confidence: 99%

Escherichia Coli Redox Mutants as Microbial Cell Factories for the Synthesis of Reduced Biochemicals

Ruiz

Almeida

Godoy

et al. 2012

Computational and Structural Biotechnology Journal

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Bioprocesses conducted under conditions with restricted O2 supply are increasingly exploited for the synthesis of reduced biochemicals using different biocatalysts. The model facultative aerobe Escherichia coli, the microbial cell factory par excellence, has elaborate sensing and signal transduction mechanisms that respond to the availability of electron acceptors and alternative carbon sources in the surrounding environment. In particular, the ArcBA and CreBC two-component signal transduction systems are largely responsible for the metabolic regulation of redox control in response to O2 availability and carbon source utilization, respectively. Significant advances in the understanding of the biochemical, genetic, and physiological duties of these regulatory systems have been achieved in recent years. This situation allowed to rationally-design novel engineering approaches that ensure optimal carbon and energy flows within central metabolism, as well as to manipulate redox homeostasis, in order to optimize the production of industrially-relevant metabolites. In particular, metabolic flux analysis provided new clues to understand the metabolic regulation mediated by the ArcBA and CreBC systems. Genetic manipulation of these regulators proved useful for designing microbial cells factories tailored for the synthesis of reduced biochemicals with added value, such as poly(3-hydroxybutyrate), under conditions with restricted O2 supply. This network-wide strategy is in contrast with traditional metabolic engineering approaches, that entail direct modification of the pathway(s) at stake, and opens new avenues for the targeted modulation of central catabolic pathways at the transcriptional level.

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Involvement of the AtoSCDAEB regulon in the high molecular weight poly-(R)-3-hydroxybutyrate biosynthesis in phaCAB+ Escherichia coli

Cited by 11 publications

References 57 publications

Enhanced co-production of hydrogen and poly-(R)-3-hydroxybutyrate by recombinant PHB producing E. coli over-expressing hydrogenase 3 and acetyl-CoA synthetase

Enhanced co-production of hydrogen and poly-(R)-3-hydroxybutyrate by recombinant PHB producing E. coli over-expressing hydrogenase 3 and acetyl-CoA synthetase

Eliminating Genes for a Two Component System Increases PHB Productivity in Cupriavidus basilensis 4G11 Under PHB Suppressing, Non-Stress Conditions

Escherichia Coli Redox Mutants as Microbial Cell Factories for the Synthesis of Reduced Biochemicals

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