Selection Method and Recombinant Microorganisms and uses Therefor
AssigneeLanzatech New Zealand Limited
InventorsMichael Koepke, Bakir Al-Sinawi
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Abstract
One or more genes in a biosynthesis pathway for a vitamin or other essential nutrient which is needed for the survival of a microorganism can be used as an effective selective marker to identify cells transformed with an exogenous nucleic acid. The microorganism does not naturally contain or express the one or more gene. This permits genetic manipulations to be performed. It permits lower cost fermentations to be performed. It permits production of the essential nutrient for subsequent commodity use.
Claims23
A process for converting CO in a gaseous CO-containing substrate into higher molecular weight products, the process comprising:
- a) passing the gaseous CO-containing substrate to a bioreactor containing a culture of carboxydotrophic acetogenic bacteria in a culture medium such that the bacteria convert the CO to higher molecule weight products, and
- b) recovering the higher molecular weight products from the bioreactor,
- wherein the carboxydotrophic acetogenic bacteria are genetically engineered to express an enzyme in a biosynthetic pathway of an essential nutrient that is absent from the culture medium, and wherein the carboxydotrophic acetogenic bacteria are prototrophic for an essential nutrient selected from the group consisting of thiamine, pantothenate, riboflavin, nicotinic acid, pyridoxine, biotin, folic acid, and cyanocobalamine, by virtue of an exogenous gene encoding the enzyme.
An isolated, genetically engineered carboxydotrophic acetogenic bacterium which is prototrophic for a vitamin selected from the group consisting of thiamine, pantothenate, riboflavin, nicotinic acid, pyridoxine, biotin, folic acid, and cyanocobalamine, by virtue of an expressed exogenous gene encoding an enzyme in a biosynthetic pathway that makes the vitamin.
A method of culturing an isolated, genetically engineered carboxydotrophic acetogenic bacterium which is prototrophic for a vitamin selected from the group consisting of thiamine, pantothenate, riboflavin, nicotinic acid, pyridoxine, biotin, folic acid, and cyanocobalamine, by virtue of an expressed exogenous gene encoding an enzyme in a biosynthetic pathway that makes the vitamin, the method comprising, growing the bacterium in a medium comprising a gaseous carbon source, wherein the carbon source comprises CO.
A method of culturing an isolated, genetically engineered carboxydotrophic acetogenic bacterium which is prototrophic for a vitamin selected from the group consisting of thiamine, pantothenate, riboflavin, nicotinic acid, pyridoxine, biotin, folic acid, and cyanocobalamine, by virtue of an expressed exogenous gene encoding an enzyme in a biosynthetic pathway that makes the vitamin, the method comprising, growing the bacterium in a medium comprising an energy source, wherein the energy source comprises CO.
A method for transferring an heterologous nucleic acid into a population of carboxydotrophic acetogenic bacteria which are auxotrophic for an essential nutrient, the method comprising:
- a) transforming the bacteria with a first nucleic acid which comprises an exogenous gene in a biosynthetic pathway of an essential nutrient, said gene operably linked to a promoter;
- and
- b) selecting for bacteria which are prototrophic for the essential nutrient among the transformed bacteria.
Dependencies are inferred from the claim text.