Carbon flux amino acid biosynthetic pathway

Metabolic flux through the anaplerotic pathways (the intermediate pathways) will be limiting for amino acid production. Overexpression or deletion of the anaplerotic enzymes may help direct the flux toward increased precursor supply for better or overproduction of the amino acid. In branching amino acid biosynthetic pathways, weakening the competing branches to direct the available precursors and net carbon flux toward the overproduction of amino acids. The pyruvate node of the TCA cycle has been engineered for increased precursor supply for lysine, glutamate. 

The effect of different amino acids supplements on the synthesis of PHB by recombinant E. coli was evaluated by Mahishi and Rawal. The study revealed that when the basal medium is supplemented with amino acids, except glycine and valine, all other amino acid supplements enhanced PHB accumulation in recombinant E. coli harboring PHB synthesizing genes from S. aureqfaciens. Cysteine, isoleucine, or methionine supplementation increased PHB accumulation by 60, 45, and 61%, respectively. Amino acid biosynthetic enzyme activities in several pathways are repressed by end produa supplementation. End product inhibition in the cysteine biosynthetic pathway controls the carbon flow due to sensitivity of serine transacetylase to cysteine. Hence, supplementation of cysteine favors a change in carbon flux that eliminates the requirement of acetyl-CoA for serine transacetylation which in turn provides more carbon source and acetyl-CoA for PHB synthesis. Degradation of methionine and isoleucine yields succinyl CoA, an intermediate of tricarboxylic acid cycle and allows more acetyl-CoA to enter the PHB biosynthetic pathway. 

L-Threonine is one of the three major amino acids produced by fermentation processes [45]. Currently, more than 4,000 tons of L-threonine are produced annually by fermentation [46]. In this section, we examine the L-threonine biosynthetic pathway and its regulation, and discuss how the carbon flux can be maximized towards L-threonine biosynthesis by metabolic engineering. The detailed description on L-threonine biosynthetic pathways and regulations involved is shown in Fig. 1. 

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