Abstract
From a gene bank of theAcinetobacter calcoaceticus genome a plasmid was isolated that complements four different classes of PQQ- mutants. Subclones of this plasmid revealed that the four corresponding PQQ genes are located on a fragment of 5 kilobases. The nucleotide sequence of this 5 kb fragment was determined and by means of Tn5 insertion mutants the reading frames of the PQQ genes could be identified. Three of the PQQ genes code for proteins of Mr 29700 (gene I), Mr 10800 (gene II) and Mr 43600 (gene III) respectively. In the DNA region where gene IV was mapped however the largest possible reading frame encodes for a polypeptide of only 24 amino acids. A possible role for this small polypeptide will be discussed. Finally we show that expression of the four PQQ genes inAcinetobacter lwoffi andEscherichia coli lead to the synthesis of the coenzyme in these organisms.
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References
Duine, JA, Frank, JznJ and vanZeeland, K, 1979. Glucose dehydrogenase fromAcinetobacter calcoaceticus: a quinoprotein. FEBS Lett 108: 186–192.
Duine, JA, Frank, Jzn J and Jongejan, JA, 1986. PQQ and quinoprotein enzymes in microbial oxidations. FEMS Microbiol Rev 32: 165–178
Devereux, J, Haeberli, P and Smithies, O, 1984. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12: 387–395
Goosen, N, Vermaas, DAM and van dePutte, P, 1987. Cloning of the genes involved in synthesis of coenzyme pyrrolo-quinoline quinone fromAcinetobacter calcoaceticus. J Bacteriol 169: 303–307
Hommes, RWJ, Postma, PW, Neyssel, OM, Tempest, DW, Dokter, P and Duine, JA, 1984. Evidence of a quinoprotein glucose dehydrogenase apoenzyme in several strains ofEscherichia coli. FEMS Microbiol Lett 24: 329–333
Kunkel, TA, 1985. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82: 488–492
Lobenstein-Verbeek, CL, Jongejan, JA, Frank, J and Duine, JA, 1984. Bovine serum amine oxidase: a mammalian enzyme having covalently bound PQQ as prosthetic group. FEBS Lett 170: 305–309
Maniatis, T, Fritsch, EF and Sambrook, J, 1982. Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N. Y.
Miller, JH, 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N. Y.
Sanger, F, Nicklen, S and Coulson, AR, 1977. DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467
van derMeer, RA, Jongejan, JA, Frank, J and Duine, JA, 1986. Hydrazone formation of 2,4-dinitrophenylhydrazine with pyrroloquinoline quinone in porcine kidney diamine oxidase. FEBS Lett 206: 111–114
van Kleef MAG and Duine JA, 1988. L-tyrosine is the precursor of PQQ biosynthesis inHyphomicrobium X. FEBS Lett in press
vanSchie, BJ, Hellingwerf, KJ, vanDijken, JP, Elferink, MGL, vanDijl, JM, Kuenen, JG and Konings, WN, 1985. Energy transduction by electron transfer via a pyrrolo-quinoline quinone dependent glucose dehydrogenase inEscherichia coli, Pseudomonas aeruginosa andAcinetobacter calcoaceticus (var lwoffi). J Bacteriol 163: 493–499
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Goosen, N., Horsman, H.P.A., Huinen, R.G.M. et al. Genes involved in the biosynthesis of PQQ fromAcinetobacter calcoaceticus . Antonie van Leeuwenhoek 56, 85–91 (1989). https://doi.org/10.1007/BF00822588
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DOI: https://doi.org/10.1007/BF00822588