Metabolic engineering for the production of dicarboxylic acids and diamines

Chae, Tong Un; Ahn, Jung Ho; Ko, Yoo-Sung; Kim, Je Woong; Lee, Jong An; Lee, Eon Hui; Lee, Sang Yup

doi:10.1016/j.ymben.2019.03.005

Cited by 107 publications

(78 citation statements)

References 100 publications

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Section: Resultsmentioning

confidence: 99%

“…Compared with other biobased fermentation methods with engineered E. coli for DCAs production 48 , our biocatalytic route provides general access to DCAs with varying chain length (C5 to C8) from different starting chemicals (e.g., cycloalkanes, cycloalkanols, or lactones). In contrast, metabolic engineering strategies require individual engineering of metabolic pathways for each DCA product 48 . In addition, with cycloalkanes or cycloalkanols as substrates, our approach gave much higher product titers than fermentation methods for production of glutaric acid 7a (1.6-6.3 g L −1 vs. 0.82 g L −1 ) 49 and suberic acid 7d (1.1-7.3 g L −1 vs. 0.254 g L −1 ) 50 , and for pimelic acid 7c, which has not been realized by metabolic pathway construction in E. coli.…”

Section: Resultsmentioning

confidence: 99%

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One-pot biocatalytic route from cycloalkanes to α,ω‐dicarboxylic acids by designed Escherichia coli consortia

Wang

Zhao

et al. 2020

Nat Commun

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Aliphatic α,ω‐dicarboxylic acids (DCAs) are a class of useful chemicals that are currently produced by energy-intensive, multistage chemical oxidations that are hazardous to the environment. Therefore, the development of environmentally friendly, safe, neutral routes to DCAs is important. We report an in vivo artificially designed biocatalytic cascade process for biotransformation of cycloalkanes to DCAs. To reduce protein expression burden and redox constraints caused by multi-enzyme expression in a single microbe, the biocatalytic pathway is divided into three basic Escherichia coli cell modules. The modules possess either redox-neutral or redox-regeneration systems and are combined to form E. coli consortia for use in biotransformations. The designed consortia of E. coli containing the modules efficiently convert cycloalkanes or cycloalkanols to DCAs without addition of exogenous coenzymes. Thus, this developed biocatalytic process provides a promising alternative to the current industrial process for manufacturing DCAs.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

One-pot biocatalytic route from cycloalkanes to α,ω‐dicarboxylic acids by designed Escherichia coli consortia

Wang

Zhao

et al. 2020

Nat Commun

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“…Since the physiological production of succinic acid is marginal in the cells, microbial cells have been engineered to develop strains that can overproduce succinic acid, and are suitable for industrial use. For example, fungi (such as Aspergillus sp., Saccharomyces cerevisiae, and Pichia kudriavzevii), rumen bacteria (Actinobacillus succiniciproducens and Mannheimia succiniciproducens), and industrial microbial strains (Corynebacterium glutamicum and Escherichia coli) have been genetically manipulated for succinic acid overproduction (Ahn et al, 2016;Chae et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Short-Term Adaptation Modulates Anaerobic Metabolic Flux to Succinate by Activating ExuT, a Novel D-Glucose Transporter in Escherichia coli

2020

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The sugar phosphotransferase system (PTS) is an essential energy-saving mechanism, particularly under anaerobic conditions. Since the PTS consumes equimolar phosphoenolpyruvate to phosphorylate each molecule of internalized glucose in the process of pyruvate generation, its absence can adversely affect the mixed acid fermentation profile and cell growth under anaerobic conditions. In this study, we report that the ptsG mutant cells of Escherichia coli K-12 strain exhibited inefficient glucose utilization, produced a significant amount of succinate, and exhibited a low growth rate. However, cells adapted soon after and started to grow rapidly in the same batch culture. As a result, the adapted ptsG cells showed the same mixed acid fermentation profiles as the wild-type cells, which was attributed to the mutation of the mlc gene, a repressor of the D-mannose PTS, another transporter for D-glucose. Similar adaptations were observed in the cells with ptsG manX and the cells with ptsI that resulted in the production of a substantial amount of succinate and fast growth rate. The genome sequencing showed the presence of null mutations in the exuR gene, which encodes a modulator of exuT-encoded non-PTS sugar transporter, in adapted ptsG manX and ptsI strains. Results from the RT-qPCR analysis and genetic test confirmed that the enhanced expression of ExuT, a non-PTS sugar transporter, was responsible for the uptake of D-glucose, increased succinate production, and fast growth of adapted cells. In conclusion, our study showed that the regulatory network of sugar transporters can be modulated by short-term adaptation and that downstream metabolic flux could be significantly determined by the choice of sugar transporters.

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“…Diacids, polyamine, and lactams are all used to produce various polyester and nylon fibers which are currently derived from petrochemicals 7,8 . In an effort to make production of these chemicals sustainable, many groups have developed engineered microbes to synthesize these precursors [9][10][11][12] .…”

Section: Introductionmentioning

confidence: 99%

Glutarate metabolism in Pseudomonas putida is regulated by two distinct glutarate sensing transcription factors

Thompson

Cruz-Morales

Krishna

et al. 2019

Preprint

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A significant bottleneck in synthetic biology involves screening the large genetically encoded libraries enabled by the latest technologies for desirable phenotypes such as chemical production. However, transcription factor based biosensors can be leveraged to screen thousands of genetic designs for optimal chemical production in engineered microbes. In this study we characterize two glutarate sensing transcription factors (CsiR and GcdR) from Pseudomonas putida. The genomic contexts of csiR homologs were analyzed and DNA binding sites were bioinformatically predicted. Both CsiR and GcdR were purified and shown to bind upstream of their coding sequencing in vitro. CsiR was shown to dissociate from DNA in vitro when exogenous glutarate was added confirming it acts as a genetic repressor. Both transcription factors and cognate promoters were then cloned into broad host range vectors to create two glutarate biosensors. Their respective sensing performance features were characterized, and more sensitive derivatives of the GcdR biosensor were created by manipulating the expression of the transcription factor. Sensor vectors were then reintroduced into P. putida and were evaluated for their ability to respond to glutarate as well as various lysine metabolites. Additionally, we developed a novel mathematical approach to describe the usable range of detection for genetically encoded biosensors which may be broadly useful to future efforts to better characterize biosensor performance.

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Metabolic engineering for the production of dicarboxylic acids and diamines

Cited by 107 publications

References 100 publications

One-pot biocatalytic route from cycloalkanes to α,ω‐dicarboxylic acids by designed Escherichia coli consortia

One-pot biocatalytic route from cycloalkanes to α,ω‐dicarboxylic acids by designed Escherichia coli consortia

Short-Term Adaptation Modulates Anaerobic Metabolic Flux to Succinate by Activating ExuT, a Novel D-Glucose Transporter in Escherichia coli

Glutarate metabolism in Pseudomonas putida is regulated by two distinct glutarate sensing transcription factors

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