Pyruvate Carboxylase and Pyruvate Carboxylase-Encoding Dna, Plasmid Containing Said Dna and Microorganism for the Production Thereof, and Methods for the Production of Products the Biosynthesis of Which Includes Oxaloacetate as Precursor, and Chromosome

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This patent describes genetically modified DNA sequences encoding improved versions of pyruvate carboxylase, an enzyme that catalyzes the formation of oxaloacetate from pyruvate. These modifications, introduced into industrial microorganisms such as Corynebacterium glutamicum or Escherichia coli, enhance the microorganisms' ability to produce valuable compounds that use oxaloacetate as a biosynthetic precursor—especially amino acids like L-lysine. The invention covers the modified gene, its expression via plasmids or genomically, the resulting microorganisms, and methods for producing increased yields of target biochemicals from fermentation processes.

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• Production of amino acids such as L-lysine, L-aspartate, L-asparagine, L-threonine, L-isoleucine, and L-methionine for food, feed, and pharmaceuticals.
• Industrial fermentation for overproduction of organic acids like succinate, malate, fumarate, citrate, and isocitrate.
• Biosynthesis of diamines (e.g., putrescine, cadaverine) for use as chemical building blocks or in polymer manufacturing.
• Improved microbial production of specialty chemicals (e.g., itaconate, ectoin, gamma-aminobutyrate, butanol, and 1-propanol).
• Biotechnological manufacturing using genetically engineered Corynebacterium, Escherichia, or yeast strains for higher yields and productivity.

BenefitsContent extracted from patent full text and abstract with AI.

• Enables higher yield (9–19% improvement demonstrated) of products derived from oxaloacetate, increasing process efficiency and profitability.
• Expands the capabilities of microbial fermentation platforms to make more diverse or valuable bioproducts.
• Reduces feedback inhibition, a common bottleneck in industrial amino acid production, leading to more robust and predictable processes.
• Applicable to a wide range of microorganisms and target products, offering flexibility for industrial biotechnology.
• Potential to lower production costs and resource usage by increasing volumetric and specific productivities.

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Patent Abstract

The invention relates to pyruvate carboxylase and a pyruvate carboxylase-encoding DNA, a plasmid containing said DNA and a microorganism for the production thereof, and to methods for the production of products the biosynthesis of which includes oxaloacetate as a precursor, and to a chromosome. The invention makes a DNA available that codes for a pyruvate carboxylase having an at least 70% sequence identity with sequence no. 1 and where the triplet that codes for threonine-343 is replaced by an alanine-encoding triplet in positions 1027-1029. In a preferred embodiment, additionally an isoleucine-encoding triplet in positions 3034-3036 of this DNA is replaced by a serine-encoding triplet. The expression of said genes results in a pyruvate carboxylase by means of which the yield in products the biosynthesis of which contains oxaloacetate as a precursor, can be increased by 9 to 19% with respect to the strain containing the native Pyc.