Microorganisms for the production of methacrylic acid

Abstract

The invention provides a non-naturally occurring microbial organism having a 2-hydroxyisobutyric acid, 3-hydroxyisobutyric acid or methacrylic acid pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in a 2-hydroxyisobutyric acid, 3-hydroxyisobutyric acid or methacrylic acid pathway. The invention additionally provides a method for producing 2-hydroxyisobutyric acid, 3-hydroxyisobutyric acid or methacrylic acid. The method can include culturing a 2-hydroxyisobutyric acid, 3-hydroxyisobutyric acid or methacrylic acid producing microbial organism expressing at least one exogenous nucleic acid encoding a 2-hydroxyisobutyric acid, 3-hydroxyisobutyric acid or methacrylic acid pathway enzyme in a sufficient amount and culturing under conditions and for a sufficient period of time to produce 2-hydroxyisobutyric acid, 3-hydroxyisobutyric acid or methacrylic acid.

1 - 240 . (canceled) 
     
     
         241 . A non-naturally occurring microbial organism, comprising one or more gene disruptions, said one or more gene disruptions occurring in genes encoding proteins or enzymes wherein said one or more gene disruptions confer increased production of 3-hydroxyisobutyric acid or methacrylic acid in said organism. 
     
     
         242 . The non-naturally occurring organism of  claim 241 , wherein production of 3-hydroxyisobutyric acid or methacrylic acid is growth-coupled. 
     
     
         243 . The non-naturally occurring organism of  claim 241 , wherein production of 3-hydroxyisobutyric acid or methacrylic acid is not growth-coupled. 
     
     
         244 . The non-naturally occurring organism of  claim 241 , wherein said one or more gene disruptions encode proteins or enzymes listed in Tables 10 or 11. 
     
     
         245 . The non-naturally occurring organism of  claim 241 , wherein said one or more gene disruptions encode proteins or enzymes selected from the group consisting of malate dehydrogenase, lactate dehydrogenase and acetaldehyde-CoA dehydrogenase. 
     
     
         246 . The non-naturally occurring organism of  claim 245 , further comprising one or more gene disruptions encoding proteins or enzymes selected from the group consisting of aspartase, pyruvate formate lyase, NAD(P) transhydrogenase, glutamate dehydrogenase, ATP synthase, phosphoenolpyruvate:pyruvate phosphotransferase system, glutamate dehydrogenase, phosphotransacetylase, acetate kinase, 6-phosphogluconolactonase, glucose 6-phosphate dehydrogenase and NADH dehydrogenase. 
     
     
         247 . The organism of  claim 241 , wherein said one or more gene disruptions comprises a deletion of said one or more genes. 
     
     
         248 . The organism of  claim 241 , wherein said cell is in a substantially anaerobic culture medium. 
     
     
         249 . A method for producing 3-hydroxyisobutyric acid or methacrylic acid comprising culturing the non-naturally occurring microbial organism of  claim 241  under conditions and for a sufficient period of time to produce 3-hydroxyisobutyric acid or methacrylic acid. 
     
     
         250 . The method of  claim 249 , wherein production of 3-hydroxyisobutyric acid or methacrylic acid is growth-coupled. 
     
     
         251 . The method of  claim 249 , wherein production of 3-hydroxyisobutyric acid or methacrylic acid is not growth-coupled. 
     
     
         252 . The method of  claim 249 , wherein said one or more gene disruptions encode proteins or enzymes listed in Tables 10 or 11. 
     
     
         253 . The organism of  claim 249 , wherein said one or more gene disruptions encode proteins or enzymes selected from the group consisting of malate dehydrogenase, lactate dehydrogenase and acetaldehyde-CoA dehydrogenase. 
     
     
         254 . The organism of  claim 253 , further comprising one or more gene disruptions encoding proteins or enzymes selected from the group consisting of aspartase, pyruvate formate lyase, NAD(P) transhydrogenase, glutamate dehydrogenase, ATP synthase, phosphoenolpyruvate:pyruvate phosphotransferase system, glutamate dehydrogenase, phosphotransacetylase, acetate kinase, 6-phosphogluconolactonase, glucose 6-phosphate dehydrogenase and NADH dehydrogenase. 
     
     
         255 . The method of  claim 249 , wherein said culturing is conducted in a substantially anaerobic culture medium. 
     
     
         256 . The method of  claim 249 , wherein said one or more gene disruptions comprises a deletion of said one or more genes.