Promoters from corynebacterium glutamicum

Abstract

Provided are native promoters comprising polynucleotides isolated from Corynebacterium glutamicum, and mutant promoters derived therefrom, which may be used to regulate, i.e., either increase or decrease, gene expression. Also provided are promoter ladders comprising a plurality of the promoters having incrementally increasing promoter activity. Also provided are host cells and recombinant vectors comprising the promoters, and methods of expressing genes of interest and producing biomolecules using the host cells.

1 - 19 . (canceled) 
     
     
         20 . A process of preparing a fine chemical, the process comprising the steps of:
 (a) culturing a microorganism that produces a fine chemical, said microorganism having increased expression of one or more genes in comparison to a counterpart starting strain by using a promoter comprising a nucleotide sequence selected from the group consisting of: SEQ ID NOs:1-8 in a suitable medium, resulting in a fermentation broth; and   (b) concentrating the fine chemical in the fermentation broth of (a) and/or in the cells of the microorganism.   
     
     
         21 . The process of  claim 20 , wherein the microorganism comprises a promoter ladder comprising at least three promoter polynucleotide sequences selected from the group consisting of SEQ ID NOs: 1-8,
 wherein said promoter ladder comprises a plurality of promoters with incrementally increasing levels of promoter activity.   
     
     
         22 . The process of  claim 21 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 1, 5 and 7. 
     
     
         23 . The process of  claim 21 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 1-2. 
     
     
         24 . The process of  claim 21 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 3, 4 and 5. 
     
     
         25 . The process of  claim 21 , wherein at least one of said promoter polynucleotide sequences is selected from the group consisting of SEQ ID NOs: 6-8. 
     
     
         26 . The process of  claim 21 , wherein each promoter of said promoter ladder is functionally linked to at least one heterologous target gene. 
     
     
         27 . The process of  claim 26 , wherein said at least one heterologous target gene is a gene that is a component of a biosynthetic pathway producing a biomolecule selected from the group consisting of amino acids, organic acids, proteins and polymers. 
     
     
         28 . The process of  claim 27 , wherein said at least one heterologous target gene is a gene that is selected from the group consisting of: aspartate-semialdehyde dehydrogenase, 4-hydroxy-tetrahydrodipicolinate synthase, dihydrodipicolinate reductase, 2,3,4,5-tetrahydropyridine-2-carboxylate N-succinyltransferase, N-succinyldiaminopimelate aminotransferase, succinyl-diaminopimelate desuccinylase, diaminopimelate epimerase, diaminopimelate decarboxylase, diaminopimelate dehydrogenase, aspartokinase lysc alpha and beta subunits, aspartate aminotransferase, glucose-specific enzyme II BC component of phosphotransferase System (PTS), glucose-6-phosphate 1-dehydrogenase, glucose-6-phosphate isomerase, transketolase, 6-phosphofructokinase 1, phosphoenolpyruvate carboxylase, pyruvate carboxylase, isocitrate dehydrogenase, phosphoenolpyruvate carboxykinase (GTP), oxaloacetate decarboxylase, homoserine kinase, homoserine dehydrogenase, and threonine synthase. 
     
     
         29 . The process of  claim 20 , wherein the microorganism comprises a recombinant polynucleotide comprising a promoter ladder comprising at least three promoter polynucleotide sequences selected from the group consisting of SEQ ID NOs: 1-8,
 wherein said promoter ladder comprises a plurality of promoters with incrementally increasing levels of promoter activity; and   wherein said promoter ladder is functionally linked to a heterologous target gene.   
     
     
         30 . The process of  claim 20 , wherein the microorganism comprises a plurality of recombinant polynucleotides comprising a promoter ladder comprising at least three promoter polynucleotide sequences selected from the group consisting of SEQ ID NOs: 1-8,
 wherein said promoter ladder comprises a plurality of promoters with incrementally increasing levels of promoter activity; and   wherein each recombinant polynucleotide comprises one promoter from the promoter ladder functionally linked to a heterologous target gene.   
     
     
         31 . The process of  claim 20 , wherein the microorganism belongs to the genus  Corynebacterium.    
     
     
         32 . The process of  claim 20 , wherein the microorganism comprises a combination of at least three promoter polynucleotide sequences selected from the group consisting of SEQ ID NOs: 1-8 each functionally linked to at least one heterologous target gene, wherein said combination of promoter polynucleotide sequences comprises a plurality of promoters with incrementally increasing levels of promoter activity. 
     
     
         33 . The process of  claim 32 , wherein said heterologous target gene is associated with a biosynthetic pathway producing a biomolecule selected from: amino acids, organic acids, flavors and fragrances, biofuels, proteins and enzymes, polymers/monomers and other biomaterials, lipids, nucleic acids, small molecule therapeutics, protein therapeutics, fine chemicals, and nutraceuticals. 
     
     
         34 . The process of  claim 32 , wherein said heterologous target gene is associated with a biosynthetic pathway producing a secondary metabolite selected from: antibiotics, alkaloids, terpenoids, and polyketides. 
     
     
         35 . The process of  claim 32 , wherein each promoter polynucleotide sequence is functionally linked to a different heterologous target gene. 
     
     
         36 . The process of  claim 32 , wherein two or more promoter polynucleotide sequences are functionally linked to at least one heterologous target gene. 
     
     
         37 . The process of  claim 36 , wherein the two or more promoter polynucleotide sequences are the same. 
     
     
         38 . The process of  claim 36 , wherein the two or more promoter polynucleotide sequences are different.