Genetically optimised microorganism for producing molecules of interest

Abstract

The invention concerns a genetically modified microorganism expressing a functional type I or II RuBisCO enzyme and a functional phosphoribulokinase (PRK), and in which the glycolysis pathway is at least partially inhibited, said microorganism being genetically modified so as to produce an exogenous molecule and/or to overproduce an endogenous molecule. According to the invention, the oxidative branch of the pentose phosphate pathway may also be at least partially inhibited. The invention also concerns the use of such a genetically modified microorganism for the production or overproduction of a molecule of interest and processes for the synthesis or bioconversion of molecules of interest.

1 . A genetically modified microorganism for the production of an exogenous molecule of interest and/or to overproduce an endogenous molecule of interest, other than a RuBisCO or phosphoribulokinase enzyme, said microorganism expressing a functional RuBisCO enzyme and a functional phosphoribulokinase (PRK), and in which the glycolysis pathway is at least partially inhibited, upstream of the production of 1,3-biphospho-D-glycerate (1,3-BPG) or upstream of the production of 3-phosphoglycerate (3PG), and downstream of the production of glyceraldehyde-3-phosphate (G3P), wherein said microorganism is genetically modified so as to produce the exogenous molecule of interest and/or to overproduce the endogenous molecule of interest, other than a RuBisCO or phosphoribulokinase enzyme. 
     
     
         2 . The genetically modified microorganism according to  claim 1 , wherein the oxidative branch of the pentose phosphate pathway is also at least partially inhibited. 
     
     
         3 . The genetically modified microorganism according to  claim 1 , wherein said microorganism is genetically modified to express a recombinant RuBisCO enzyme and/or PRK. 
     
     
         4 . The genetically modified microorganism according to  claim 2 , wherein said microorganism is genetically modified to inhibit the oxidative branch of the pentose phosphate pathway upstream of ribulose-5-phosphate production. 
     
     
         5 . The genetically modified microorganism according to  claim 1 , wherein the exogenous molecule and/or the endogenous molecule is selected from amino acids, peptides, proteins, vitamins, sterols, flavonoids, terpenes, terpenoids, fatty acids, polyols and organic acids. 
     
     
         6 . The genetically modified microorganism according to  claim 1 , wherein said microorganism is a eukaryotic cell or a prokaryotic cell. 
     
     
         7 . The genetically modified microorganism according to  claim 1 , wherein the expression of the gene encoding glyceraldehyde 3-phosphate dehydrogenase is at least partially inhibited. 
     
     
         8 . The genetically modified microorganism according to  claim 1 , wherein the expression of the gene encoding phosphoglycerate kinase is at least partially inhibited. 
     
     
         9 . The genetically modified microorganism according to  claim 7 , wherein the expression of the gene encoding glucose-6-phosphate dehydrogenase or 6-phosphogluconolactonase or 6-phosphogluconate dehydrogenase is at least partially inhibited. 
     
     
         10 . The genetically modified microorganism according to  claim 1 , wherein said microorganism is a yeast of the genus  Saccharomyces cerevisiae  genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the TDH1, TDH2 and/or TDH3 gene is at least partially inhibited. 
     
     
         11 . The genetically modified microorganism according to  claim 1 , wherein said microorganism is a  Saccharomyces cerevisiae  yeast genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the PGK1 gene is at least partially inhibited. 
     
     
         12 . The genetically modified microorganism according to  claim 10 , wherein the expression of the ZWF1 gene is at least partially inhibited. 
     
     
         13 . The genetically modified microorganism according to  claim 1 , wherein said microorganism is a filamentous fungus of the genus  Aspergillus  genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the pgk and gsdA genes is at least partially inhibited. 
     
     
         14 . The genetically modified microorganism according to  claim 1 , wherein said microorganism is an  E. coli  bacterium genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the gapA and/or pgk gene, and optionally the zwf gene, is at least partially inhibited. 
     
     
         15 .- 16 . (canceled) 
     
     
         17 . A biotechnological process for producing at least one molecule of interest, wherein it comprises a step of culturing a genetically modified microorganism as defined in  claim 1 , under conditions allowing the synthesis or bioconversion by said microorganism of said molecule of interest, and optionally a step of recovering and/or purifying said molecule of interest. 
     
     
         18 . The biotechnological process according to  claim 17 , wherein the molecule of interest is selected from amino acids, peptides, proteins, vitamins, sterols, flavonoids, terpenes, terpenoids, fatty acids, polyols and organic acids. 
     
     
         19 . The biotechnological process according to  claim 17 , wherein the molecule of interest is selected glutamate, citrate, itaconate or GABA. 
     
     
         20 . The biotechnological process according to  claim 17 , wherein the microorganism is genetically modified to express at least one enzyme involved in the bioconversion or synthesis of said molecule of interest. 
     
     
         21 . The biotechnological process according to  claim 17 , wherein the microorganism is genetically modified to at least partially inhibit an enzyme involved in the degradation of said molecule of interest. 
     
     
         22 . A process for producing a molecule of interest comprising (i) inserting at least one sequence encoding an enzyme involved in the synthesis or bioconversion of said molecule of interest into a recombinant microorganism as defined in  claim 1 , (ii) culturing said microorganism under conditions allowing expression of said enzyme and optionally (iii) recovering and/or purifying said molecule of interest. 
     
     
         23 . A process for producing a molecule of interest comprising (i) inhibiting the expression of at least one gene encoding an enzyme involved in the degradation of said molecule of interest in a recombinant microorganism as defined in  claim 1 , (ii) culturing said microorganism under conditions allowing expression of said enzyme and optionally (iii) recovering and/or purifying said molecule of interest. 
     
     
         24 . The genetically modified microorganism of  claim 1 , wherein said microorganism is a eukaryotic cell selected from yeasts, fungi and microalgae. 
     
     
         25 . The genetically modified microorganism of  claim 1 , wherein said microorganism is a bacterium.