Phosphoenolpyruvate carboxylase mutant and application thereof
By performing dual-site mutations (R528H and S828P) on phosphoenolpyruvate carboxylase, the problem of competitive consumption of pyruvate by the intracellular TCA cycle bypass pathway was solved, thereby improving the production capacity of L-valine and achieving a significant yield increase.
CN120905200APending Publication Date: 2025-11-07ANHUI HUAHENG BIOTECH CO LTD +1
2 Cites 0 Cited by
Patent Information
- Application Number
- CN202511054124.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-29
- Publication Date
- 2025-11-07
AI Technical Summary
Technical Problem
During the biosynthesis of L-valine in Escherichia coli, the competitive consumption of phosphoenolpyruvate by alternative pathways such as the intracellular TCA cycle limits the accumulation of pyruvate, resulting in insufficient L-valine production capacity.
Method used
By performing dual mutations at R528H and S828P on phosphoenolpyruvate carboxylase (ppc), its catalytic activity is reduced, thereby weakening competition from the bypass pathway and increasing the supply of pyruvate precursors.
Benefits of technology
It significantly increased L-valine production by 27.7% while maintaining normal cell growth requirements, thus promoting the development of a new generation of high-yield L-valine strains.
✦ Generated by Eureka AI based on patent content.
Abstract
The invention relates to a phosphoenolpyruvate carboxylase mutant and application thereof, in particular to the phosphoenolpyruvate carboxylase mutant, compared with an amino acid sequence of wild phosphoenolpyruvate carboxylase protein as shown in SEQ ID NO: 1, the phosphoenolpyruvate carboxylase mutant has R528H and S828P substitutions, and the amino acid sequence of the phosphoenolpyruvate carboxylase mutant is shown in SEQ ID NO: 1. And the activity of phosphoenolpyruvate carboxylase is reduced. By mutating phosphoenolpyruvate carboxylase, the protein expression activity of phosphoenolpyruvate carboxylase is reduced, bypass pathway competition is weakened, and pyruvic acid precursor supply is enhanced, so that the production capacity and biomass of valine engineering strains are further improved. Through fermentation verification, compared with an initial bacterium Sval024, the L-valine yield of a recombinant strain containing the ppc (R528H / S828P) mutant gene is increased by 27.7%, and the recombinant strain has important industrial application value for developing the L-valine high-yield strain.
Need to check novelty before this filing date? Find Prior Art