PRODUCTION OF MUCONIC ACID FROM GENETICALLY ENGINEERED MICROORGANISMS
This present invention is in the field of producing renewable chemical feedstocks using biocatalysts that have been genetically engineered to increase their ability to convert renewable carbon resources into useful compounds. More specifically, the present invention provides a process for producing muconic acid form renewable carbon resources using a genetically modified organism.
1 . A genetically engineered Escherichia coli in which the PEP-dependent phosphotransferase system and the GalP-based system for glucose import are eliminated, and which comprises an exogenous glucose facilitator system comprising a protein encoded by an exogenous glf genes.
2 . A genetically engineered Escherichia coli as in claim 1 further comprising a pck gene coding for phosphoenol pyruvate carboxykinase enzyme with increased activity.
3 . A genetically engineered Escherichia coli as in claim 1 further comprising a deletion in the pykA gene coding for pyruvate kinase enzyme activity.
4 . A genetically engineered Escherichia coli as in claim 1 further comprising a deletion in the pykF gene coding for pyruvate kinase enzyme activity.
5 . A genetically engineered Escherichia coli as in claim 1 further comprising one or more exogenous genes selected from a group consisting of aroZ, qa-4, asbF, aroY, quiC and catAX, wherein said one or more exogenous genes code for proteins functional in a muconic acid pathway.
6 . A genetically engineered Escherichia coli as in claim 1 further comprising one or more exogenous genes selected from a group consisting of aroB, aroD, aroF, aroG, aroH, tktA, talB, rpe, and rpi, wherein said one or more exogenous genes code for proteins functional in a shikimic acid pathway.
7 . A genetically engineered Escherichia coli as in claim 1 wherein the activity of a negative regulator protein of aromatic amino acid biosynthesis encoded by a tyrR gene or its homolog is substantially reduced or eliminated.
8 . A genetically engineered Escherichia coli as in claim 1 further comprising an aroG* gene which codes for a DAHP synthase enzyme that is substantially resistant to inhibition by phenylalanine.
9 . A genetically engineered Escherichia coli of claim 8 further comprising an aroG* gene that codes for a DAHP synthase enzyme that is selected from a group consisting of aroG*20-893, aroG*20-897 aroG*20-899, aroG*20-901, aroG*111, aroG*211, aroG*212, aroG*311, aroG*312, aroG*411, aroG*412, and aroG*511.
10 . A genetically engineered Escherichia coli of claim 1 further comprising an exogenous gene that codes for a QuiC enzyme of a bacterium of the genus Acinetobacter , or a homolog of said QuiC enzyme.