Microbial Strain Optimization for Plant Protein Synthesis

Producing enzymes from microorganisms through controlled glucose supplementation. The method employs glycosylamine as a feedstock to enhance enzyme production in microorganisms like Trichoderma reesei. The glycosylamine is added to the culture medium, where it is converted into glucose through enzymatic hydrolysis. This controlled glucose supply prevents catabolite repression while maintaining optimal enzyme expression levels. The glycosylamine feedstock can be precisely controlled in concentration and flow rate to achieve the desired enzyme production levels.

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Genetically engineered microorganisms that can co-produce proteins and chemicals from CO2 through continuous fermentation of industrial substrates. The engineered microorganisms have tandem repeat genes in their genomes, enabling the production of complex proteins and chemicals with multiple carbon atoms. These microorganisms can be engineered to produce a wide range of products, including proteins, chemicals, biomass, and biopolymers, from CO2, CO, and H2 through continuous fermentation processes. The engineered microorganisms can be used as a platform for sustainable chemical production, protein synthesis, and biomass conversion.

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Method for enhancing protein production in microorganisms like Bacillus subtilis through the use of a heterologous FKBP (FK506 binding protein) enzyme. The method involves expressing the heterologous FKBP in the microorganism, particularly in a Bacillus subtilis strain lacking extracellular proteases, to improve protein secretion and yield. The heterologous FKBP is engineered to have prolyl isomerization activity, which enables efficient protein folding and secretion. The engineered microorganism is cultured under optimal conditions, and the target protein is recovered and purified. This approach enables significant productivity gains in protein production by leveraging the natural binding function of the heterologous FKBP enzyme.

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A genetically engineered yeast strain for producing high-value protein concentrates through controlled methanol fermentation. The engineered strain enables the production of protein concentrates with improved nutritional profile and yield compared to traditional Pichia pastoris strains. The engineered strain is engineered to express the necessary enzymes for methanol metabolism, and the process involves precise genetic modification to introduce the desired enzymes. The engineered yeast can be optimized through controlled mutagenesis to enhance protein production and improve fermentation efficiency.

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Bacterial strains of Bifidobacterium, Lactobacillus, Lactococcus, Lactobacillus acidophilus, Lactiplantibacillus plantarum, and Lacticaseibacillus rhamnosus that selectively break down plant proteins and enhance amino acid and peptide availability. These strains, when administered in food products or dietary supplements, exhibit enhanced proteolytic activity compared to their non-probiotic counterparts, resulting in improved protein digestibility and bioavailability. The strains specifically target plant proteins such as soy, pea, fava bean, chickpea, and lentil proteins, demonstrating their efficacy in improving protein utilization in both humans and animals.

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Yeast cells engineered to produce recombinant proteins with reduced exopolysaccharide production, achieved through manipulating the mannosyltransferase pathway. The cells selectively underexpress key enzymes in this pathway, specifically beta-mannosyltransferase 1 and beta-mannosyltransferase 2, which are responsible for the production of complex polysaccharides in yeast. By reducing the expression of these enzymes, the cells can produce recombinant proteins with lower levels of exopolysaccharide impurities while maintaining high protein yields.

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Fungal strains engineered to significantly enhance heterologous protein expression levels through targeted genetic modifications. The engineered strains contain mutations in the GSH1 and Not4 genes, which are crucial for GSH1 activity and Not4 function in the host cell. These mutations lead to increased expression levels of the target protein, enabling higher yields and improved purification characteristics compared to the parent strain. The engineered strains can be used for commercial protein production, particularly for applications requiring high levels of heterologous protein.

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A host cell engineered to utilize non-natural carbon sources for recombinant protein production. The engineered cell expresses a fusion protein containing a catalytic domain of a glycosyl hydrolase and a surface-bound anchoring domain of a glycosylphosphatidylinositol (GPI)-anchored protein. The cell's growth rate is enhanced when fed non-natural carbon sources, such as sucrose, maltose, fructose, or high-fructose corn syrup, compared to standard glucose. This cell line enables the production of recombinant proteins using these alternative carbon sources, particularly for therapeutic proteins and biopharmaceuticals.

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A genetically engineered microorganism for continuous production of targeted chemical products and exogenous proteins from CO2, CO, and H2 through a gas fermentation process. The engineered organism, a Cl-fixing bacterium, produces both a high-value protein (e.g., ethylene) and a chemical (e.g., ethylene) through continuous fermentation, with the gas substrate being a renewable energy source. The engineered microorganism can be engineered to produce tandem-repeating proteins and secreted chemicals, enabling the simultaneous production of both valuable products.

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A high-yielding strain of Fusarium venenatum, a species used to produce mycelium protein, that can produce more protein and grow faster than existing strains. The strain, named Fusarium venenatum TB03, has mutations in genes involved in polyadenylation, transposase function, and cellulose synthesis. These mutations improve protein accumulation in the mycelium. The TB03 strain provides a starting point for optimizing mycelium protein production using Fusarium venenatum.

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High-yield Fusarium venenatum mutant strain for producing mycelium protein. The mutant strain has non-synonymous mutations in genes like zinc cluster protein, aspartic protease, ATF4 activation transcription factor, and cell differentiation protein. These mutations increase protein yield when growing Fusarium venenatum on glucose substrate. The mutant strain is deposited as Fusarium venenatum TBO1 (CGMCC 20740).

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Producing high-value nutritional products through gaseous fermentation of microorganisms using waste streams from industrial processes. The method involves cultivating microorganisms that can convert inorganic carbon into organic compounds through chemoautotrophic metabolism, such as CO2, CH4, and H2. These microorganisms are engineered to grow on waste substrates like CO2, CH4, and H2, and produce valuable products like proteins, vitamins, and enzymes. The process enables the production of biomass and food-grade ingredients from waste streams, with the added benefit of carbon-neutral production.

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High-yield production of recombinant proteins in Pichia pastoris yeast cells through integrated expression cassettes. The approach enables efficient expression of heterologous proteins by combining multiple expression cassettes with diverse promoters, each carrying a separate copy of the heterologous gene sequence. The integrated cassettes are integrated into the yeast genome in a non-homologous manner, allowing for higher copy numbers and improved stability compared to traditional homologous integration methods. This enables the production of high-titer recombinant proteins in large-scale fermentation cultures.

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A strain of Fusarium mycelial protein-producing strain that enables high protein production through fermentation with various nitrogen sources. The strain, identified as Fusarium venenatum TB01, exhibits enhanced mycelial protein yields when grown on different nitrogen substrates, including inorganic and organic sources. This strain enables the production of mycelial protein with a protein content of over 40%, with optimal yields achieved when using sulfuric acid, urea, or organic nitrogen sources.

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A method to enhance nitrogen fixation in non-legume crops by genetically engineering bacteria to improve their ability to fix atmospheric nitrogen. The engineered bacteria incorporate a nitrogen-fixing gene with high sequence identity to a known nitrogen-fixing gene, allowing them to fix nitrogen even in the presence of supplemental fertilizers. The engineered bacteria can be applied to crops like corn, wheat, and soybeans through seed inoculation, enabling farmers to achieve nitrogen fixation while reducing fertilizer use.

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A method for increasing protein production in microbial biomass production through co-culture with methane-oxidizing bacteria. The approach involves using a specific strain of Klebsiella pneumonia 1-17 as a co-culture partner for methane-oxidizing bacteria, which can utilize methane homologues and bacterial byproducts. The co-culture benefits from the methane-oxidizing bacteria's ability to convert natural gas into biomass while the Klebsiella pneumonia 1-17 strain produces proteins, amino acids, and polysaccharides through its lysis process. This co-culture approach enables enhanced protein production compared to traditional pure culture systems, particularly when using methane as a substrate.

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A Corynebacterium strain engineered to produce L-lysine through enhanced protein expression. The engineered strain specifically expresses a protein with the amino acid sequence of L-lysine, which is absent in the original strain. This engineered protein is characterized by its increased activity compared to the original strain's endogenous L-lysine production. The engineered strain can be cultivated in standard media, and L-lysine can be recovered from the culture medium. This engineered strain offers a significant improvement in L-lysine production compared to the original strain, with an enhanced protein activity compared to the endogenous L-lysine production.

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A strain of bacteria R. daidae that selectively colonizes eastern goat plants, enhancing nitrogen fixation and plant growth through a symbiotic relationship. The strain exhibits high specificity to the goat host, promoting enhanced nitrogen fixation and plant development. It demonstrates rapid growth rates and is resistant to antibiotics, making it a valuable tool for agricultural biopreparation. The strain's ability to form symbiotic relationships with goat plants enables efficient nitrogen fixation, improving plant biomass and quality.

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