Method for Producing Vectors Containing a Gene That Codes for an Enzyme Having Reduced or Switched-Off Feedback Inhibition, and Use Thereof for Producing Amino Acids and Nucleotides

Simple SummaryContent extracted from patent full text and abstract with AI.

This invention discloses a method to create gene vectors that encode enzymes with reduced or eliminated feedback inhibition. Such vectors are developed by mutating genes in vitro, inserting them into vectors, and using metabolite-sensing microorganisms to identify those that result in elevated target metabolite production. The identified vectors code for enzymes that are less sensitive to feedback inhibition, allowing microorganisms transformed with these vectors to overproduce valuable compounds like amino acids or nucleotides.

Use CasesContent extracted from patent full text and abstract with AI.

• Industrial-scale production of amino acids (e.g., L-lysine, L-glutamate) for food additives, feed supplements, and pharmaceuticals.
• Biosynthesis of nucleotides for use in nutritional supplements and biotechnology processes.
• Development of engineered microbial strains with enhanced productivity for fermentation-based manufacturing processes.
• Production of non-natural or rare amino acids as intermediates for specialty chemicals or pharmaceuticals.
• Generation of microbial strains for research into metabolic regulation and synthetic biology.

BenefitsContent extracted from patent full text and abstract with AI.

• Enables higher yields of important biomolecules such as amino acids and nucleotides through microorganism fermentation.
• Eliminates or reduces the need for analog-based selection methods, leading to more reliable and efficient strain improvement.
• Allows for rapid, high-throughput identification of desirable enzyme mutants using metabolite sensors and fluorescence-based screening.
• Improves the efficiency and accuracy of developing feedback-resistant enzyme variants compared to traditional mutagenesis approaches.
• Facilitates the creation of tailored strains for industry, reducing production costs and increasing scalability.

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Patent Abstract

The invention relates to a method for producing vectors containing a gene that codes for an enzyme having reduced or switched-off feedback inhibition, and use thereof for producing amino acids and nucleotides. According to the invention genes that code for feedback-inhibited enzymes are mutagenised in vitro, in a further step are ligated into vectors, in a further step each vector is transformed into a microorganism that contains a metabolite sensor, which at increased metabolite concentration causes the synthesis of a fluorescent protein, after which microorganisms having an increased or the highest fluorescence are selected, and the vectors that contain a gene coding for an enzyme having reduced or switched-off feedback inhibition are isolated from microorganisms having an increased or the highest fluorescence.