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metabolic engineering of Escherichia coli and Corynebacterium glutamicum strains by expression of plant enzyme anthranilic acid methyltransferase1 (AAMT1). Optimizing the key enzyme anthranilic acid (ANT) methyltransferase1 (AAMT1) expression, increasing the direct precursor ANT supply, and enhancing the intracellular availability and salvage of the cofactor S-adenosyl-L-methionine required by AAMT1, results in improved methyl anthranilate (MANT) production in both engineered microorganisms. In situ two-phase extractive fermentation using tributyrin as an extractant is developed to overcome MANT toxicity. Fed-batch cultures of the final engineered Escherichia coli and Corynebacterium glutamicum strains in two-phase cultivation mode lead to the production of 4.47 and 5.74 g/l MANT, respectively, in minimal media containing glucose. Metabolic network related to MANT biosynthesis from glucose in Escherichia coli and Corynebacterium glutamicum, as well as metabolic engineering strategies, methods, overview |
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