Summary
A 6940 by Klebsiella pneumoniae chromosomal DNA fragment, containing genes involved in pyrroloquinoline quinone (PQQ) biosynthesis, was sequenced. Six open reading frames, pqqA, pqqB, pqqC, pqqD, pqqE and pqqF were identified in the pqq operon, which coded for polypeptides of 2764 (23 amino acids), 33 464, 28 986, 10 436, 42 881 and 83 616 Da, respectively. The transcription startpoint was mapped by primer extension analysis, upstream of qgqA, and promoter boxes could be identified. The gene products of pqqB, pqqC, pqqE and pqqF were detected in maxi-cells and the molecular weights of the proteins corresponded with the molecular weights deduced from the nucleotide sequence. The gene products of pqqA, pqqB, pqqC, pqqD and pqqE show 49%–64% identity in amino acid sequence with those of pqqIV, pqqV, pqqI, pqqII and pqqIII respectively in the cloned pqq cluster of Acinetobacter calcoaceticus. The 84 kDa protein encoded by pgqF, which is not present in the cloned pqq cluster of A. calcoaceticus but which is essential for PQQ biosynthesis in K. pneumoniae and Escherichia coli, seems to belong to a family of proteases.
Similar content being viewed by others
References
Aflholter JA, Fried VA, Roth RA (1988) Human insulin-degrading enzyme shares structural and functional homologies with E. coli protease III. Science 242:1415–1418
Aiba H, Adhya L, de Crombrugghe B (1981) Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem 256:11905–11910
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Biville F, Turlin E, Gasser F (1989) Cloning and genetic analysis of six pyrroloquinoline quinone biosynthesis genes in Methylobacterium organophilum DSM 760. J Gen Microbiol 135:2917–2929
Borck K, Beggs JD, Brammar WJ, Hopkins AS, Murray NE (1976) The construction in vitro of transducing derivatives of phage lambda. Mol Gen Genet 146:199–207
Boyd AC, Sherratt DJ (1986) Polar mobilization of the Escherichia coli chromosome by the ColEl transfer origin. Mol Gen Genet 203:496–504
Clarke L, Carbon J (1978) Functional expression of cloned yeast DNA in Escherichia coli: Specific complementation of argininosuccinate lyase (argH) mutations. J Mol Biol 120:517–532
Cowan ST (1975) Family I. Enterobacteriaceae. In: Buchanan RE, Gibbons NE (eds) Bergey's manual of determinative bacteriology, 8th edition. The Williams and Wilkins Company, Baltimore, pp 290–293
Curtis SJ, Epstein W (1975) Phosphorylation of D-glucose in Escherichia coli mutants defective in glucosephosphotransferase, mannosephosphotransferase and glucokinase. J Bacteriol 122:1189–1199
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Duine JA (1991) Quinoproteins: enzymes containing the quinonoid cofactor pyrroloquinoline quinone, topaquinone or tryptophantryptophan quinone. Eur J Biochem 200:271–284
Duine JA, Frank J (1980) The prosthetic group of methanol dehydrogenase. Purification and some of its properties. Biochem J 187:213–219
Duine JA, Jongejan JA (1989) Enzymes with pyrroloquinoline quinone as cofactor. Annu Rev Biochem 58:403–426
Duine JA, Frank J, Van Zeeland K (1979) Glucose dehydrogenase from Acinetobacter calcoaceticus: A “quinoprotein”. FEBS Lett 108:443–446
Duine JA, Frank J, Jongejan JA (1987) Enzymology of quinoproteins. Adv Enzymol 59:169–212
Finch PW, Wilson RE, Brown K, Hickson ID, Emmerson PT (1986) Complete nucleotide sequence of the Escherichia coli ptr gene encoding Protease III. Nucleic Acids Res 14:7695–7703
Goosen N, Vermaas DAM, van de Putte P (1987) Cloning of the genes involved in synthesis of the coenzyme pyrroloquinoline quinone from Acinetobacter calcoaceticus. J Bacteriol 169:303–307
Goosen N, Horsman HPA, Huinen RGM, de Groot A, van de Putte P (1989a) Genes involved in the biosynthesis of PQQ from Acinetobacter calcoaceticus. In: Duine JA, Jongejan JA (eds) PQQ and quinoproteins. Proceedings of the 1st International Symposium on PQQ and Quinoproteins, Delft, The Netherlands, 1988. Kluwer Academic, Dordrecht, pp 169–176
Goosen N, Horsman HPA, Huinen RGM, van de Putte P (1989b) Acinetobacter calcoaceticus genes involved in biosynthesis of the coenzyme pyrroloquinoline-quinone: Nucleotide sequence and expression in Escherichia coli K-12. J Bacteriol 171:447–455
Groen B, van Kleef MAG, Duine JA (1986) Quinohaemoprotein alcohol dehydrogenase apoenzyme from Pseudomonas testosteroni. Biochem J 234:611–615
Harley CB, Reynolds RP (1987) Analysis of E. coli promoter sequences. Nucleic Acids Res 15:2343–2361
Higgins DG, Sharp PM (1988) Clustal; A package for performing multiple sequence alignment on a micro computer. Gene 73:237–244
Hommes RWJ, Postma PW, Neijssel OM, Tempest DW, Dokter P, Duine JA (1984) Evidence for a glucose dehydrogenase apoenzyme in several strains of Escherichia coli. FEMS Microbiol Lett 24:329–333
Houck DR, Hanners JL, Unkefer CJ (1988) Biosynthesis of pyrroloquinoline quinone. Identification of biosynthetic precursors using 13C labelling and NMR spectroscopy. J Am Chem Soc 110:6920–6921
Kushner SR (1978) Methods for transformation of Escherichia coli. In: Boyer HB, Nicosia S (eds) Genetic engineering. Elsevier, Amsterdam, pp 17–23
Lazdunski AM (1989) Peptidases and proteases of Escherichia coli and Salmonella typhimurium. FEMS Microbiol Rev 63:265–276
Maniatis T, Fritsch EF, Sambrook L (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Messing J, Crea R, Seeburg PH (1981) A system for shotgun DNA sequencing. Nucleic Acids Res 9:309–321
Meulenberg JJM, Loenen WAM, Sellink E, Postma PW (1990a) A general method for the transfer of plasmid-borne mutant alleles to the chromosome of Klebsiella pneumoniae using bacteriophage lambda: Transfer of pqq genes. Mol Gen Genet 220:481–484
Meulenberg JJM, Sellink E, Loenen WAM, Riegman NH, van Kleef M, Postma PW (1990b) Cloning of Klebsiella pneumoniae pqq genes and PQQ biosynthesis in Escherichia coli. FEMS Microbiol Lett 71:337–344
Neijssel OM, Tempest DW, Postma PW, Duine JA, Frank Jzn J (1983) Glucose metabolism by K+-limited Klebsiella aerogenes, evidence for the involvement of a quinoprotein glucose dehydrogenase. FEMS Microbiol Lett 20:35–39
Nilsson B, Uhlen M, Josephson S, Gatenbeck S, Philipson L (1983) An improved positive selection vector constructed by oligonucleotide mediated mutagenesis. Nucleic Acids Res 11:8019–8029
Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Sancar A, Wharton RP, Selzer S, Kacinski BM, Clark MD, Rupp WD (1981) Identification of the uvrA gene product. J Mol Biol 148:45–62
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Scholte BJ, Schuitema ARJ, Postma PW (1982) Characterization of factor IIIGlc in catabolite repression resistant (crr) mutants of Salmonella typhimurium. J Bacteriol 149:576–586
Schulte U, Arretz M, Schneider H, Tropschug M, Wachter E, Neupert W, Weiss H (1989) A family of mitochondrial proteins involved in bioenergetics and biogenesis. Nature 339:147–149
Shine J, Dalgarno L (1974) 3′-Terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346
Simon R, Quandt J, Klipp W (1989) New derivatives of transposon Tn5 suitable for mobilization of replicons, generation of operon fusions and induction of genes in Gram-negative bacteria. Gene 80:161–169
Van Kleef MAG, Duine JA (1988a) L-tyrosine is the precursor of PQQ biosynthesis in Hyphomicrobium X. FEBS Lett 237:91–97
Van Kleef MAG, Duine JA (1988b) A search for intermediates in the bacterial biosynthesis of PQQ. Biofactors 1:297–302
Van Schie BJ, van Dijken JP, Kuenen JG (1984) Non coordinated synthesis of glucose dehydrogenase and its prosthetic group PQQ in Acinetobacter and Pseudomonas species. FEMS Microbiol Lett 24:133–138
Way C, Davis MA, Morisato D, Roberts DE, Kleckner N (1984) New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene 32:369–379
Witte C, Jensen RE, Yaffe MR, Schatz G (1988) masl, a gene essential for yeast mitochondrial assembly, encodes a subunit of the mitochondrial processing protease. EMBO J 7:1439–1447
Author information
Authors and Affiliations
Additional information
Communicated by J. Lengeler
Rights and permissions
About this article
Cite this article
Meulenberg, J.J.M., Sellink, E., Riegman, N.H. et al. Nucleotide sequence and structure of the Klebsiella pneumoniae pqq operon. Molec. Gen. Genet. 232, 284–294 (1992). https://doi.org/10.1007/BF00280008
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00280008