Processes to prepare elongated 2-ketoacids and C5-C10 compounds therefrom via genetic modifications to microbial metabolic pathways
A C6-C10, gene modification technology, applied in the field of microorganisms of isopropylmalate synthase, can solve the problem that microorganisms cannot produce
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[0070] Preparation of a genetically modified isopropylmalate synthase with enhanced catalytic activity towards 2-ketohexanoate and 2-ketooctanoate.
[0071] During the biosynthesis of 2-ketononanoic acid via the recursive activity of the "+1" pathway of the leucine biosynthetic pathway, isopropylmalate synthase traps and condenses acetyl-CoA and 2-ketoacids of various lengths. For efficient biosynthesis of 2-ketononanoic acid, isopropylmalate synthase is expected to efficiently capture 2-ketobutyrate, 2-ketovalerate, 2-ketohexanoate, 2-ketoheptanoate and 2-ketoheptanoate with acetyl-CoA. and / or 2-ketooctanoic acid, resulting in the corresponding 2-alkylmalic acid product ( figure 1 Intermediate II) in. The native isopropylmalate synthase of E. coli and other microorganisms is less efficient at capturing longer unnatural 2-ketoacid substrates. To increase the activity of native isopropylmalate synthase to capture longer 2-keto acids for catalysis, the active site of isopropy...
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