Pyruvate Carboxylase Having a Feedback-Resistant Mutation, Encoding Dna, Plasmides, and Microorganism for the Production Thereof, and Methods for the Production of Products the Biosynthesis of Which Includes Oxaloacetate as Precursor

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This patent describes a genetically engineered version of the enzyme pyruvate carboxylase, specifically modified to resist feedback inhibition. The modification involves substituting the amino acid isoleucine at position 1012 with serine in the enzyme's sequence. This enzyme, along with its encoding DNA, plasmids, and transformed microorganisms, is used to boost the production of biochemicals whose biosynthesis starts from oxaloacetate, such as lysine, glutamate, and other amino acids, as well as certain organic acids and diamines. The invention results in up to a 7% increase in yield for these target products compared to strains with the native (unmodified) enzyme.

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• Industrial microbial production of amino acids such as L-lysine, L-aspartate, L-glutamate, L-methionine, etc.
• Biotechnological manufacturing of organic acids/intermediates from the citric acid cycle (e.g., succinate, malate, fumarate, citrate, isocitrate, 2-oxoglutarate).
• Production of diamines (e.g., cadaverine, putrescine) for use in polymers and specialty chemicals.
• Biosynthesis of specialty compounds like itaconate, gamma-aminobutyrate, ectoine, 1-propanol, butanol, and hydroxyproline.
• Use in engineered strains of Corynebacterium, Escherichia coli, yeast, and other industrial microorganisms for improved fermentation efficiency.

BenefitsContent extracted from patent full text and abstract with AI.

• Increases yield (up to 7%) of oxaloacetate-derived products in microbial fermentation processes.
• Reduces feedback inhibition in central metabolic pathways, enabling more robust production under industrial conditions.
• Enables more efficient and cost-effective production of essential and specialty amino acids, organic acids, and platform chemicals.
• Genetic modification can be applied to various host organisms widely used in biotechnology, increasing its industrial applicability.
• Provides a method for improved gene expression and enzyme activity, allowing further enhancements to fermentation processes.

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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 isoleucine-1012 is replaced by a serine-encoding triplet in positions 3034-3036. The expression of said gene results in a pyruvate carboxylase by means of which the yield of the reaction of metabolic products derived from oxaloacetate is by 7% higher with respect to the strain containing the native Pyc.