Microorganisms and methods for the continuous co-production of high-value, specialized proteins and chemical products from C1-substrates
Inventors
Simpson, Sean Dennis • Koepke, Michael • Clomburg, James MacAllister • Gramling, Juniper Dee • Jones, Stephanie Rhianon • Jung, Jaeyoung • Politano, Timothy James • Tappel, Ryan Christopher
Assignees
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Publication Number
US-12291734-B2
Publication Date
2025-05-06
Expiration Date
Abstract
Microorganisms are genetically engineered to continuously co-produce amino acids, high-value, specialized proteins, microbial biomass, chemicals, or any combination thereof by microbial fermentation, particularly by microbial fermentation of a gaseous substrate. The microorganisms are C1-fixing. The production of ethylene, microbial biomass, and heterologous high-value, specialized proteins can be improved. This can be improved by varying promoters or nutrient limiting means.
Core Innovation
The invention provides a process for continuous co-production of a chemical product and at least one exogenous gene product using a continuous bioreactor. A recombinant C1-fixing microorganism selected from Clostridium autoethanogenum and Cupriavidus necator is introduced with a gaseous substrate comprising one or more of CO, CO2, and H2 and a liquid growth medium, and continuous culturing generates a gas fermentation broth comprising the chemical product, the exogenous gene product, and microbial biomass.
The process continuously removes a portion of the gas fermentation broth in a first stream, continuously removes at least one chemical product in a second stream, and continuously recovers the exogenous gene product from the microbial biomass from the first stream. The exogenous gene product is squid ring teeth (SRT) protein, and the chemical product is selected from acetone, ethanol, ethylene, isoprene, and isopropanol.
The disclosure further describes continuous gas-fermentation co-production using recombinant C1-fixing microorganisms in continuous CSTR settings for producing chemical products while generating detectable recombinant tandem-repeat proteins. Example contexts include continuous ethylene production from CO2 with H2 as an energy source in a CSTR, and a bubble-generating reactor system for enhancing gas-liquid mass transfer using a sparger and perforated plate.
Claims Coverage
The independent claim covers continuous co-production in a continuous bioreactor using a recombinant C1-fixing microorganism, with split-stream removal of the chemical product and recovery of an exogenous gene product from microbial biomass. One independent claim is present, with dependent refinements to the chemical product identity and the exogenous gene product.
Continuous bioreactor co-production with recombinant C1-fixing microorganism
providing a continuous bioreactor; introducing to the bioreactor a recombinant C1-fixing microorganism selected from Clostridium autoethanogenum and Cupriavidus necator capable of co-producing at least one chemical product and at least one exogenous gene product, a gaseous substrate comprising one or more of CO, CO2, and H2, and a liquid growth medium; continuously culturing the recombinant C1-fixing microorganism thereby generating a gas fermentation broth comprising the at least one chemical product, the at least one exogenous gene product, and microbial biomass;
First-stream portion removal and biomass-based exogenous gene product recovery
continuously removing a portion of the gas fermentation broth in a first stream; continuously recovering the at least one exogenous gene product from the microbial biomass from the first stream;
Second-stream chemical product removal while maintaining continuous co-production
continuously removing the at least one chemical product in a second stream;
SRT exogenous gene product coupled with selected chemical products
wherein the at least one exogenous gene product is squid ring teeth (SRT) protein and wherein the at least one chemical product is selected from acetone, ethanol, ethylene, isoprene, and isopropanol.
The coverage centers on continuous co-production in a continuous bioreactor using a recombinant C1-fixing microorganism fed with a gaseous substrate, generating a gas fermentation broth containing both a chemical product and squid ring teeth (SRT) protein, and using split-stream removal with recovery of the exogenous gene product from microbial biomass.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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