Engineering a synthetic pathway for maleate in Escherichia coli

Noda, Shinshi; Shirai, Toshiaki; Mori, Yutaro; Oyama, Sachiko; Kondo, Akihiko

doi:10.1038/s41467-017-01233-9

Cited by 40 publications

(39 citation statements)

References 45 publications

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“…4 By introducing related genes into Escherichia coli, we can e ciently produce chorismate derivatives, such as ccMA and maleate (a valuable unsaturated C4 carboxylic acid), from glucose. 21,22,23 The central skeleton structure of ccMA is consistent with that of 1,3-butadiene, so twostep repeated ccMA decarboxylation can produce 1,3-butadiene.…”

Section: Introductionmentioning

confidence: 72%

Direct 1,3-butadiene biosynthesis in Escherichia coli via a tailored ferulic acid decarboxylase mutant

Mori

Noda

Shirai

et al. 2020

Preprint

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The C4 unsaturated compound 1,3-butadiene is an important monomer in synthetic rubber and engineering plastic production. However, microorganisms cannot directly produce 1,3-butadiene using glucose as a renewable carbon source via biological processes. This study constructed a novel artificial metabolic pathway for 1,3-butadiene production from glucose in Escherichia coli by combining the cis,cis-muconic acid (ccMA)-producing pathway and tailored ferulic acid decarboxylase mutants. The rational enzyme design of the substrate-binding site by computational simulation improved 1,3-butadiene-producing ccMA decarboxylation with a small mutant library. We found that changing dissolved oxygen and controlling pH were important factors for 1,3-butadiene production. Using dissolved oxygen–stat fed-batch fermentation in a 1-L jar fermenter, we could produce 2.1 g L− 1 of 1,3-butadiene. These results indicated that using a rational enzyme design, we can produce unnatural/nonbiological compounds (those that are made from petroleum and cannot be produced by microorganisms) using glucose as a renewable carbon source.

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Section: Introductionmentioning

confidence: 72%

Direct 1,3-butadiene biosynthesis in Escherichia coli via a tailored ferulic acid decarboxylase mutant

Mori

Noda

Shirai

et al. 2020

Preprint

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“…To further enhance the arbutin production, we attempted to identify and improve the rate-limiting step in the synthetic pathway of arbutin production. On the basis of previous study for enhanced shikimate pathway [ 17 – 19 ], four categories of genes were selected from the gluconeogenesis pathway, pentose phosphate pathway, shikimate pathway and “chorismate-arbutin” pathway. The first group, PEP synthase encoded by ppsA was generally used to improve the precursor PEP.…”

Section: Resultsmentioning

confidence: 99%

“…Numerous studies reporting deregulation of the feedback inhibition, enhancement of the shikimate pathway, and increased accumulation of precursors PEP and E4P have been attempted to increase the production of target biologicals of shikimate pathway [ 23 – 26 ]. As competitive branches, the disruption of aromatic amino acid (Tyr, Trp, and Phe) biosynthesis pathway might contribute to the improvement of chorismate derivatives [ 27 , 28 ].…”

Section: Discussionmentioning

confidence: 99%

Enhanced biosynthesis of arbutin by engineering shikimate pathway in Pseudomonas chlororaphis P3

Wang

Bilal

et al. 2018

Microb Cell Fact

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BackgroundArbutin is a plant-derived glycoside with potential antioxidant, antibacterial and anti-inflammatory activities. Currently, it is mainly produced by plant extraction or enzymatic processes, which suffers from expensive processing cost and low product yield. Metabolic engineering of microbes is an increasingly powerful method for the high-level production of valuable biologicals. Since Pseudomonas chlororaphis has been widely engineered as a phenazine-producing platform organism due to its well-characterized genetics and physiology, and faster growth rate using glycerol as a renewable carbon source, it can also be engineered as the cell factory using strong shikimate pathway on the basis of synthetic biology.ResultsIn this work, a plasmid-free biosynthetic pathway was constructed in P. chlororaphis P3 for elevated biosynthesis of arbutin from sustainable carbon sources. The arbutin biosynthetic pathway was expressed under the native promoter Pphz using chromosomal integration. Instead of being plasmid and inducer dependent, the metabolic engineering approach used to fine-tune the biosynthetic pathway significantly enhanced the arbutin production with a 22.4-fold increase. On the basis of medium factor optimization and mixed fed-batch fermentation of glucose and 4-hydroxybenzoic acid, the engineered P. chlororaphis P3-Ar5 strain led to the highest arbutin production of 6.79 g/L with the productivity of 0.094 g/L/h, with a 54-fold improvement over the initial strain.ConclusionsThe results suggested that the construction of plasmid-free synthetic pathway displays a high potential for improved biosynthesis of arbutin and other shikimate pathway derived biologicals in P. chlororaphis.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-1022-8) contains supplementary material, which is available to authorized users.

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“…In addition to fueling biosynthesis, synthetic pathways have been built for the biosynthesis of various chemical compounds, such as succinic acid [39], 2,4-dihydroxybutyric acid [40], and maleate [41], and the engineered bio-factories have already been commercialized for the food industry.…”

Section: Pathway Constructionmentioning

confidence: 99%

Advancement of Metabolic Engineering Assisted by Synthetic Biology

Lee

2018

Catalysts

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Synthetic biology has undergone dramatic advancements for over a decade, during which it has expanded our understanding on the systems of life and opened new avenues for microbial engineering. Many biotechnological and computational methods have been developed for the construction of synthetic systems. Achievements in synthetic biology have been widely adopted in metabolic engineering, a field aimed at engineering micro-organisms to produce substances of interest. However, the engineering of metabolic systems requires dynamic redistribution of cellular resources, the creation of novel metabolic pathways, and optimal regulation of the pathways to achieve higher production titers. Thus, the design principles and tools developed in synthetic biology have been employed to create novel and flexible metabolic pathways and to optimize metabolic fluxes to increase the cells’ capability to act as production factories. In this review, we introduce synthetic biology tools and their applications to microbial cell factory constructions.

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Engineering a synthetic pathway for maleate in Escherichia coli

Cited by 40 publications

References 45 publications

Direct 1,3-butadiene biosynthesis in Escherichia coli via a tailored ferulic acid decarboxylase mutant

Direct 1,3-butadiene biosynthesis in Escherichia coli via a tailored ferulic acid decarboxylase mutant

Enhanced biosynthesis of arbutin by engineering shikimate pathway in Pseudomonas chlororaphis P3

Advancement of Metabolic Engineering Assisted by Synthetic Biology

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