Catechol biosynthesis from glucose in Escherichia coli anthranilate-overproducer strains by heterologous expression of anthranilate 1,2-dioxygenase from Pseudomonas aeruginosa PAO1

Abstract

BackgroundThe aromatic compound catechol is used as a precursor of chemical products having multiple applications. This compound is currently manufactured by chemical synthesis from petroleum-derived raw materials. The capacity to produce catechol is naturally present in several microbial species. This knowledge has been applied to the generation of recombinant Escherichia coli strains that can produce catechol from simple carbon sources.ResultsSeveral strains derived from E. coli W3110 trpD9923, a mutant that overproduces anthranilate, were modified by transforming them with an expression plasmid carrying genes encoding anthranilate 1,2-dioxygenase from Pseudomonas aeruginosa PAO1. The additional expression of genes encoding a feedback inhibition resistant version of 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase and transketolase from E. coli, was also evaluated. Generated strains were characterized in complex or minimal medium in shake-flask and fed-batch bioreactor cultures and incubation temperatures ranging from 37 to 28°C. These experiments enabled the identification of culture conditions for the production of 4.47 g/L of catechol with strain W3110 trpD9923, expressing 1,2-dioxygenase, DAHP synthase and transketolase. When considering the amount of glucose consumed, a yield of 16% was calculated, corresponding to 42% of the theoretical maximum as determined by elementary node flux analysis.ConclusionsThis work demonstrates the feasibility of applying metabolic engineering for generating E. coli strains for the production of catechol from glucose via anthranilate. These results are a starting point to further optimize environmentally-compatible production capacity for catechol and derived compounds.

Metabolic Engineering of Shikimic Acid Biosynthesis Pathway for the Production of Shikimic Acid and Its Branched Products in Microorganisms: Advances and Prospects

“…The main methods used to produce catechol are phenol and m-diisopropylbenzene oxidation and coal tar distillation [ 21 ]. However, due to the complex chemical synthesis process, serious pollution, requirement of expensive catalysts, and other such problems, these methods have been phased out.…”

“…Catechol is mostly produced via chemical synthesis using raw materials extracted from petroleum. The ability to produce catechol naturally is observed only in a small number of microbial species, and acquiring the required carbon source is a complex process; hence, the requirements of industrial production are not met [ 21 ]. Fortunately, with the further development of metabolic engineering, recombinant bacteria have been constructed for the production of catechol from simple carbon sources [ 93 ].…”

“…Significant achievements have been made in SA production via these methods [ 20 ]. The microbial metabolic engineering method is also of great significance for QA, GA, and catechol production, thereby effectively meeting the market demand for these products [ 21 , 22 , 23 ].…”

Production of Melanins With Recombinant Microorganisms

“…To further improve this process, metabolic engineering was evaluated to generate a strain with the capacity of generating catechol melanin from a simple carbon source. The strategy that was followed is based on employing an engineered E. coli strain that can produce catechol from a simple carbon source (Balderas-Hernández et al, 2014). Strain E. coli W3110 trpD9923 is a mutant in the L-tryptophan biosynthetic pathway that overproduces the intermediate anthranilate (Yanofsky et al, 1971).…”

Catechol-Loading Nanofibrous Membranes for Eco-Friendly Iron Nutrition of Plants

“…Several distinct organisms (plants and microorganisms as well as mollusks) synthesize and use catechol, although for different purposes [35,36,37,38,39], but always exploiting the Fe-binding capacity of this molecule. Catechol, for example, is part of the phenolic compounds that are the major components of root exudates released in response to iron (Fe) deficiency in Strategy I plants [40,41], and the catechol motif is present in several other soluble phenolic compounds of root exudates, such as coumarins (esculetin, fraxetin, and sideretin) [33,42,43] and flavonoids (quercetin, catechin) [44,45,46], where this structural moiety is specifically devoted to iron binding.…”