Abstract
Metabolic engineering is an enabling technology for producing chemicals, pharmaceuticals, and fuels in a green and sustainable manner. Enzymes are key catalysts for metabolic reactions for the synthesis of target product. In this chapter, we initially discuss enzyme research in metabolic engineering, fueled by screening the enzymes of key biochemical pathways from different organisms for an enhanced production. Next, the optimization of key pathway enzymes by feedback inhibition removal, catalytic efficiency improvement, and substrate specificity alteration are discussed. Finally, assembling of the key pathway enzymes for balancing and strengthening synthetic pathways is discussed, including fusion expression of key enzymes, synthetic scaffold-guided enzyme co-localization, and compartmentalization engineering. The systematic summary and discussion of screening, optimization and assembling of key pathway enzymes in metabolic engineering may facilitate metabolic engineers to further combine protein engineering with metabolic engineering for eliminating rate-limiting steps for improved production.
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