Corynebacterium glutamicum genes encoding phosphoenolpyruvate: sugar phosphotransferase system proteins

Abstract

Isolated nucleic acid molecules, designated PTS nucleic acid molecules, which encode novel PTS proteins from Corynebacterium glutamicum are described. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing PTS nucleic acid molecules, and host cells into which the expression vectors have been introduced. The invention still further provides isolated PTS proteins, mutated PTS proteins, fusion proteins, antigenic peptides and methods for the improvement of production of a desired compound from C. glutamicum based on genetic engineering of PTS genes in this organism.

1 . An isolated polypeptide selected from the group consisting of: 
 a) an isolated polypeptide comprising the amino acid sequence of SEQ ID NO:18;    b) an isolated polypeptide comprising a naturally occurring allelic variant of a polypeptide comprising the amino acid sequence of SEQ ID NO:18;    c) an isolated polypeptide which is encoded by a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:17;    d) an isolated polypeptide which is encoded by a nucleic acid molecule comprising a nucleotide sequence which is at least 90% identical to the entire nucleotide sequence of SEQ ID NO:17;    e) an isolated polypeptide comprising an amino acid sequence which is at least 90% identical to the entire amino acid sequence of SEQ ID NO:18;    f) an isolated polypeptide comprising a fragment of a polypeptide comprising the amino acid sequence of SEQ ID NO:18, wherein said polypeptide fragment maintains a biological activity of the polypeptide comprising the amino sequence; and    g) an isolated polypeptide which is encoded by a nucleic acid molecule which hybridizes to the complement of the nucleotide sequence of SEQ ID NO:17 in 6× sodium chloride/sodium citrate (SSC) at 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 50-65° C., or a complement thereof.    
     
     
         2 . The isolated polypeptide of  claim 1 , further comprising heterologous amino acid sequences.  
     
     
         3 . The isolated polypeptide of  claim 1 , wherein the polypeptide has a phosphoenolpyruvate-protein phosphotransferase activity.  
     
     
         4 . A method for producing a fine chemical, comprising culturing a recombinant host cell capable of expressing the polypeptide of  claim 1 .  
     
     
         5 . The method of  claim 4 , wherein said method further comprises the step of recovering the fine chemical from said culture.  
     
     
         6 . The method of  claim 4 , wherein said cell belongs to the genus  Corynebacterium  or  Brevibacterium.    
     
     
         7 . The method of  claim 4 , wherein said cell is selected from the group consisting of  Corynebacterium glutamicum, Corynebacterium herculis, Corynebacterium lilium, Corynebacterium acetoacidophilum, Corynebacterium acetoglutamicum, Corynebacterium acetophilum, Corynebacterium ammoniagenes, Corynebacterium fujiokense, Corynebacterium nitrilophilus, Brevibacterium ammoniagenes, Brevibacterium butanicum, Brevibacterium divaricatum, Brevibacterium flavum, Brevibacterium healii, Brevibacterium ketoglutamicum, Brevibacterium ketosoreductum, Brevibacterium lactofermentum, Brevibacterium linens, Brevibacterium paraffinolyticum , and those strains set forth in Table 3.  
     
     
         8 . The method of  claim 4 , wherein expression of the polypeptide results in modulation of production of said fine chemical.  
     
     
         9 . The method of  claim 4 , wherein said fine chemical is selected from the group consisting of organic acids, proteinogenic and nonproteinogenic amino acids, purine and pyrimidine bases, nucleosides, nucleotides, lipids, saturated and unsaturated fatty acids, diols, carbohydrates, aromatic compounds, vitamins, cofactors, polyketides, and enzymes.  
     
     
         10 . The method of  claim 4 , wherein said fine chemical is an amino acid.  
     
     
         11 . The method of  claim 10 , wherein the amino acid is selected from the group consisting of lysine, glutamate, glutamine, alanine, aspartate, glycine, serine, threonine, methionine, cysteine, valine, leucine, isoleucine, arginine, proline, histidine, tyrosine, phenylalanine, and tryptophan.