Arginine supplementation to improve efficiency in gas fermenting acetogens
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.
1. A method for growing a C1-fixing microorganism, comprising flowing a gaseous substrate comprising one or more of CO, CO2, and H2 to a bioreactor containing a culture of a C1-fixing microorganism in a liquid nutrient medium;
wherein arginine is provided to the culture as a sole nitrogen source; wherein the C1-fixing microorganism is a recombinant, genetically modified microorganism comprising an arginine metabolism pathway; and wherein the C1-fixing microorganism is Clostridium autoethanogenum, Clostridium ljungdahlii , or Clostridium ragsdalei.
2. The method of claim 1 , wherein the arginine metabolism pathway comprises an arginine deiminase pathway and/or an arginine decarboxylase pathway, wherein the arginine deiminase pathway comprises one or more enzymes selected from the group consisting of arginine deiminase (EC 3.5.3.6), ornithine carbamoyltransferase (EC 2.1.3.3), and carbamate kinase (EC 2.7.2.2), and wherein the arginine decarboxylase pathway comprises one or more enzymes selected from the group consisting of arginine decarboxylase (EC 4.1.1.19), agmatine deiminase (EC 3.5.3.12), putrescine carbamoyl transferase (EC 2.1.3.6), and carbamate kinase (EC 2.7.2.2).
3. The method of claim 2 , wherein the arginine metabolism pathway comprises one or more of an overexpressed endogenous enzyme, a mutant endogenous enzyme, or an exogenous enzyme.
4. The method of claim 1 , wherein the concentration of arginine in the bioreactor is at least 330 mg/L.
5. The method of claim 1 , wherein the culture produces no acetate.
6. The method of claim 1 , wherein arginine is catabolized by the arginine metabolism pathway to produce ammonium, wherein the C1-fixing microorganism utilizes ammonium as the nitrogen source.
7. The method of claim 1 , wherein the C1-fixing microorganism comprises a disruptive mutation in an arginine:ornithine transporter.
8. The method of claim 1 , wherein the C1-fixing microorganism comprises one or more genetic modifications to the arginine metabolism pathway, wherein the one or more genetic modifications are selected from the group consisting of (i) a disruptive mutation of regulatory elements and (ii) a replacement of operator binding sites or native promoters with constitutive or synthetic promoters.
9. The method of claim 8 , wherein the disruptive mutation is a knock-out of arginine repressor argR.
10. The method of claim 8 , wherein the replacement is a replacement of an arginine deiminase pathway operon promoter with a constitutive or synthetic promoter.