Production of <i>p</i>-Aminobenzoic acid by metabolically engineered <i>Escherichia coli</i>

Koma, Daisuke; Yamanaka, Hayato; Moriyoshi, Kunihiko; Sakai, Kiyofumi; Masuda, Takaya; Satō, Yoshihiro; Toida, Kozo; Ohmoto, Takashi

doi:10.1080/09168451.2014.878222

Cited by 34 publications

(35 citation statements)

References 29 publications

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“…Given that pABA production is kinetically limited, in a situation of high flux into the shikimate pathway during faster glucose metabolism, chorismate could be accumulating, leading to an overflow into PEA (a similar flux overflow resulting in an accumulation of PHE and TYR has been reported when producing pABA in E. coli (Koma et al, 2014)). …”

Section: Metabolic Bottlenecks Of Carbon-source Utilization and Aromamentioning

confidence: 76%

“…Recently also the production of pABA in E. coli has been reported, reaching maximum titers of 4.8 g/L at a yield of 21% (Koma et al, 2014). This was accomplished by employing the E. coli aroF fbr (DAHP synthase) and pabC (aminodeoxychorismate lyase) genes in combination with a Corynebacterium efficiens pabAB (aminodeoxychorismate synthase).…”

Section: Bio-based Production Of Phba and Pabamentioning

confidence: 99%

“…However, the production was moderate (250 µM), raising the question, if ABZ2 might be currently limiting pABA production in yeast. Recently overproduction was reported in E. coli (4.8 g/L) where also the choice of the bacterial analogues to ABZ1 and ABZ2 (pabA & pabB / pabAB and pabC) was shown to be crucial (Koma et al, 2014).…”

Section: K229lmentioning

confidence: 99%

“…The prokaryotic functional analogous pabAB & pabC are another set of enzymes which seem very promising (Koma et al, 2014) for enhancing pABA formation. Finally, the design of a fusion protein of the aminodeoxychorismate synthase & aminodeoxychorismate lyase (e.g.…”

Section: Future Perspectivesmentioning

confidence: 99%

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Engineering yeast shikimate pathway towards production of aromatics: rational design of a chassis cell using systems and synthetic biology

Averesch¹

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Aromatic building blocks are amongst the most important bulk feedstocks in the chemical industry. As these compounds are commonly derived from petrochemistry, obtaining them is becoming more and more a matter of costs and sustainability. Biochemistry gives rise to a wealth of compounds that can potentially replace current petroleum-based chemicals or be used for novel materials. The aromatic compounds para-aminobenzoic acid (pABA) and para-hydroxybenzoic acid (pHBA) and the aromatics derived compound cis,cismuconic acid (ccMA) can be precursors for, but are not limited to, terephthalic or/and adipic acid. These are essential feedstocks for the production of PET and nylon. The three compounds can be derived from the shikimate pathway, an anabolic pathway leading to the biosynthesis of aromatic amino acids, present in certain prokaryotes and eukaryotes, including fungi. By combination of metabolic modelling with genetic engineering, a microbial production system based on the yeast Saccharomyces cerevisiae can be designed, which effectively channels flux into the target compounds.In order to develop a competitive bio-based process, yields, titers and rates need to be maximized. While productivity or rates in a process can be altered using genetic engineering, carbon yield, and pathway feasibility are stoichiometrically and thermodynamically predetermined. Both limitations need to be considered when designing a microbial production system. For formation of adipic acid and precursors many bio-based routes exists. More rational than just picking one for in vivo studies rather all available biochemical pathways were examined in silico using metabolic modelling. To compare theoretical yields and reaction thermodynamics an interface that allowed network-embedded thermodynamic analysis of elementary flux modes was developed. This allowed distinguishing between thermodynamically feasible and infeasible flux distributions. Feasible maximum theoretical product carbon yields were substantially different in E. coli and S. cerevisiae metabolic models and ranged from 32% to 92%. Further, many pathways appeared to be restricted by a thermodynamic equilibrium lying on the substrate side, some even infeasible.The only routes that deliver significant product yields and were thermodynamically favoured were shikimate pathway based. Being currently of strong scientific interest, recent implementations of these pathways in E. coli and S. cerevisiae were evaluated and strain construction strategies optimized, using the concept of constrained minimal cut-sets.Especially in S. cerevisiae a single non-obvious knock-out target allowed coupling of growth to product formation; in particular, the deletion of the pyruvate kinase reaction resulted in a minimum yield constraint of 28%.Though unique to shikimate pathway, this strategy is transferrable to other products which are derived from chorismate and also involve the formation of pyruvate as a by-product. This applies to pHBA. With further optimizations, the strategy was applied in...

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Section: Metabolic Bottlenecks Of Carbon-source Utilization and Aromamentioning

confidence: 76%

Section: Bio-based Production Of Phba and Pabamentioning

confidence: 99%

Section: K229lmentioning

confidence: 99%

Section: Future Perspectivesmentioning

confidence: 99%

See 2 more Smart Citations

Engineering yeast shikimate pathway towards production of aromatics: rational design of a chassis cell using systems and synthetic biology

Averesch¹

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“…These are used in sunscreens, dyes, liquid crystal polymers, polyurethanes and food additives and have also the potential to act as building blocks for aromatic polymers. 281 Even though purely synthesised from petrochemicals to date, there is potential for the bio-production of pHBA and pABA as microbes such as E.…”

Section: 3-butanediol (23-bdo) Is An Interesting Example To Study mentioning

confidence: 99%

Understanding extracellular electron transport of industrial microorganisms and optimization for production application

Kracke¹

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Microbial electrosynthesis (MES) and electro-fermentation are novel approaches to increase microbial production by stimulating the metabolism of the cells electrically. The technique is still in its infancy and while already hyped as promising method to convert CO 2 or cheap carbon sources and electrical energy into valuable chemicals and fuels, little is known about its true potential. This project uses a combination of in silico and in vivo approaches to gather novel information about the benefits and limitations of microbial electrosynthesis as well as its fundamental mechanisms.Understanding microbial electron transport mechanisms is the key to optimization of any bioelectrochemical technology. Therefore, this work analyses the different electron transport chains that nature offers for organisms such as metal respiring bacteria and acetogens, but also standard biotechnological organisms currently used in bioproduction.Special focus lies on the essential connection of redox and energy metabolism, which is often ignored when studying bioelectrochemical systems. The possibility of extracellular electron exchange at different points in each organism is discussed regarding required redox potentials and effect on cellular redox and energy levels. Key compounds such as electron carriers (e.g. cytochromes, ferredoxins, quinones, flavins) are identified and analysed regarding their possible role in electrode-microbe-interactions.Based on these findings a stoichiometric network analysis is performed analysing the effect of electrical stimulation on the microbial metabolism theoretically. Escherichia coli is used as model organism for production with the aim to identify target processes for MES.For the first time, 20 different valuable products were screened for their potential to show increased yields during anaerobic electrically enhanced fermentation. Surprisingly it was found that an increase in product formation by electrical enhancement is not necessarily dependent on the degree of reduction of the product but rather the metabolic pathway it is derived from. Contrary to the usual assumption a reduced product would always require electron supply by a cathode this study shows that a complex metabolic analysis is needed to identify the overall redox state of each process. A variety of beneficial processes is presented with product yield increases of maximal +36% in reductive and +84% in oxidative fermentations and final theoretical product yields up to 100%. This includes compounds that are already produced at industrial scale such as succinic acid, ii lysine and diaminopentane as well as potential novel bio-commodities such as isoprene, para-hydroxybenzoic acid and para-aminobenzoic acid. Furthermore, it is shown that the way of electron transport has major impact on achievable biomass and product yields. The coupling of electron transport to energy conservation could be identified as crucial for most processes.The in vivo approach of this project introduces the development of a standardised reactor platfor...

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Anthranilic Acid and Aniline

2023

Pathway Design for Industrial Fermentation

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Production of p-Aminobenzoic acid by metabolically engineered Escherichia coli

Cited by 34 publications

References 29 publications

Engineering yeast shikimate pathway towards production of aromatics: rational design of a chassis cell using systems and synthetic biology

Engineering yeast shikimate pathway towards production of aromatics: rational design of a chassis cell using systems and synthetic biology

Understanding extracellular electron transport of industrial microorganisms and optimization for production application

Anthranilic Acid and Aniline

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