Arginine supplementation to improve efficiency in gas fermenting acetogens
AssigneeLanzatech New Zealand Limited
InventorsKaspar Valgepea, Michael Koepke, James Bruce Yarnton Haycock Behrendorff, Esteban Marcellin, Lars K. Nielsen, Renato de S.P. Lemgruber
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Abstract
The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways.
Claims25
A method for increasing the production of at least one ATP-intensive product, the method comprising;
- a. flowing a gaseous C1-containing substrate to a bioreactor containing a culture of a C1-fixing microorganism in a liquid nutrient media;
- and
- b. fermenting the culture to produce at least one product;
- wherein arginine is provided to the culture in excess of the cellular requirement of the culture;
- and wherein the C1-fixing microorganism comprises an arginine metabolism pathway.
A method for improving the sustainability of a fermentation process, the method comprising:
- a. flowing a gaseous C1-containing substrate to a bioreactor containing a culture of at least one C1-fixing bacterium comprising at least one of an arginine deiminase pathway or an arginine decarboxylase pathway in a liquid nutrient media;
- b. fermenting the culture to produce at least one product;
- and
- c. providing arginine to the culture in excess of the cellular requirement of the culture, such that arginine is catabolized by the arginine deiminase pathway to produce ammonium, wherein the bacterium utilizes ammonium as a nitrogen source.
A method for improving the efficiency of a fermentation process, the method comprising providing arginine as the sole nitrogen source for a C1-fixing microorganism.
A genetically engineered C1-fixing microorganism comprising an optimized arginine deiminase pathway.
A method for improving the efficiency of arginine incorporation into metabolism, the method comprising culturing a genetically engineered C1-fixing microorganism comprising one or more genetic modifications selected from the group consisting of:
- i. a disruptive mutation in an arginine transporter;
- ii. overexpression of one or more endogenous enzymes selected from the group consisting of arginine deiminase (EC 3.5.3.6), ornithine carbomyltransferase (putrescine carbomyltransferase) (EC 2.1.3.3), carbamate kinase (EC 2.7.2.2), omithine racemase (EC 5.1.1.12), omithine aminomutase (EC 5.4.3.5), 2,4-diaminopentanoate dehydrogenase (EC 1.4.1.12), and 2-amino-4-oxopentanoate thiolase (EC 2.3.1.B10);
- iii. expression of one or more mutated endogenous enzymes selected from the group consisting of arginine deiminase (EC 3.5.3.6), ornithine carbomyltransferase (putrescine carbomyltransferase) (EC 2.1.3.3), carbamate kinase (EC 2.7.2.2), ornithine racemase (EC 5.1.1.12), ornithine aminomutase (EC 5.4.3.5), 2,4-diaminopentanoate dehydrogenase (EC 1.4.1.12), and 2-amino-4-oxopentanoate thiolase (EC 2.3.1.B10);
- and
- iv. expression of one or more exogenous enzymes selected from the group consisting of arginine deiminase (EC 3.5.3.6), ornithine carbomyltransferase (putrescine carbomyltransferase) (EC 2.1.3.3), carbamate kinase (EC 2.7.2.2), ornithine racemase (EC 5.1.1.12), ornithine aminomutase (EC 5.4.3.5), 2,4-diaminopentanoate dehydrogenase (EC 1.4.1.12), and 2-amino-4-oxopentanoate thiolase (EC 2.3.1.B10)
A method for improving efficiency of arginine co-utilization with one or more gaseous substrates selected from the group consisting of CO, H2 and CO2, the method comprising culturing a genetically engineered C1-fixing bacterium comprising one or more genetic modifications, wherein the one or more genetic modifications are selected from the group consisting of (i) disruptive mutation of regulatory elements and (ii) replacement of operator binding sites or native promoters with constitutive or synthetic promoters.
Dependencies are inferred from the claim text.