+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Cloning, sequence analysis, expression and inactivation of the Corynebacterium glutamicum pta-ack operon encoding phosphotransacetylase and acetate kinase



Cloning, sequence analysis, expression and inactivation of the Corynebacterium glutamicum pta-ack operon encoding phosphotransacetylase and acetate kinase



Microbiology 145 pt 2: 3-13



The Corynebacterium glutamicum ack and pta genes encoding the acetate-activating enzymes acetate kinase and phosphotransacetylase were isolated, subcloned on a plasmid and re-introduced into Corynebacterium glutamicum. Relative to the wild-type, the recombinant strains showed about tenfold higher specific activities of both enzymes. Sequence analysis of a 3657 bp DNA fragment revealed that the ack and pta genes are contiguous in the corynebacterial chromosome, with pta upstream and the last nucleotide of the pta stop codon (TAA) overlapping the first of the ack start codon (ATG). The predicted gene product of pta consists of 329 amino acids (Mr 35,242), that of ack consists of 397 amino acids (Mr 43,098) and the amino acid sequences of the two polypeptides show up to 60% (phosphotransacetylase) and 53% (acetate kinase) identity in comparison with respective enzymes from other organisms. Northern (RNA) blot hybridizations using pta- and ack-specific probes and transcriptional cat fusion experiments revealed that the two genes are transcribed as a 2.5 kb bicistronic mRNA and that the expression of this operon is induced when Corynebacterium glutamicum grows on acetate instead of glucose as a carbon source. Directed inactivation of the chromosomal pta and ack genes led to the absence of detectable phosphotransacetylase and acetate kinase activity in the respective mutants and to their inability to grow on acetate. These data indicate that no isoenzymes of acetate kinase and phosphotransacetylase are present in Corynebacterium glutamicum and that a functional acetate kinase/phosphotransacetylase pathway is essential for growth of this organism on acetate. Reprinted by permission of the publisher.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 009796171

Download citation: RISBibTeXText

PMID: 10075432

DOI: 10.1099/13500872-145-2-503


Related references

Cloning, sequence analysis, expression, and inactivation of the Corynebacterium glutamicum icd gene encoding isocitrate dehydrogenase and biochemical characterization of the enzyme. Journal of Bacteriology 177(3): 774-782, 1995

Cloning, sequence analysis, and hyperexpression of the genes encoding phosphotransacetylase and acetate kinase from Methanosarcina thermophila. Journal of Bacteriology 175(21): 6822-6829, 1993

Cloning and heterologous expression of the genes encoding acetate kinase and phosphotransacetylase from Lactobacillus sanfranciscensis. Abstracts of the General Meeting of the American Society for Microbiology 99: 503, 1999

Cloning, sequencing, and expression of genes encoding phosphotransacetylase and acetate kinase from Clostridium acetobutylicum ATCC 824. Applied and Environmental Microbiology 62(8): 2758-2766, 1996

Identification, sequence analysis, and expression of a Corynebacterium glutamicum gene cluster encoding the three glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase, and triosephosphate isomerase. Journal of Bacteriology 174(19): 6076-6086, 1992

Cloning of the pyruvate kinase gene (pyk) of Corynebacterium glutamicum and site-specific inactivation of pyk in a lysine-producing Corynebacterium lactofermentum strain. Applied and Environmental Microbiology 60(7): 2494-2500, 1994

Cloning and structural analysis of ribosomal rna operon of corynebacterium glutamicum. Doklady Akademii Nauk SSSR 308(6): 1476-1477, 1989

Cloning and structural analysis of the operon of ribosomal RNA of Corynebacterium glutamicum. Doklady Akademii Nauk SSSR 308(6): 1476-1477, 1989

Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiology 140: 1817-1828, 1994

Molecular cloning nucleotide sequence and fine structural analysis of the corynebacterium glutamicum fda gene structural comparison of corynebacterium glutamicum fructose 1 6 bisphosphate aldolase to class i and class ii aldolases. Molecular Microbiology 3(11): 1625-1638, 1989

Identification and characterization of the ackA (acetate kinase A)-pta (phosphotransacetylase) operon and complementation analysis of acetate utilization by an ackA-pta deletion mutation of Escherichia coli. Journal of Biochemistry (Tokyo) 116(4): 916-922, 1994

Identification and characterization of the ackA (acetate kinase A)-pta (phosphotransacetylase) operon and complementation analysis of acetate utilization by an ackA-pta deletion mutant of Escherichia coli. Journal of Biochemistry 116(4): 916-922, 1994

Nucleotide sequence and fine structural analysis of the Corynebacterium glutamicum hom-thrB operon. Molecular Microbiology 2(1): 63-72, 1988

Lrp of Corynebacterium glutamicum controls expression of the brnFE operon encoding the export system for L-methionine and branched-chain amino acids. Journal of Biotechnology 158(4): 231-241, 2012