Summary
Gluconobacter oxydans contains pyrroloquinoline quinone-dependent glucose dehydrogenase (GDH). Two isogenic G. oxydans strains, P1 and P2, which differ in their substrate specificity with respect to oxidation of sugars have been analysed. P1 can oxidize only d-glucose, whereas P2 is also capable of the oxidation of the disaccharide maltose. To investigate the nature of this maltose-oxidizing property we cloned the gene encoding GDH from P2. Expression of P2 gdh in P1 enables the latter strain to oxidize maltose, indicating that a mutation in the P2 gdh gene is responsible for the change in substrate specificity. This mutation could be ascribed to a 1 by substitution resulting in the replacement of His 787 by Asn.
Similar content being viewed by others
References
Buchert J, Viikari L (1988) Oxidative d-xylose metabolism of Gluconobacter oxydans. Appl Microbiol Biotechnol 29:375–379
Cleton-Jansen AM, Goosen N, Wenzel TJ, Van de Putte P (1988a) Cloning of the gene encoding quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus: evidence for the presence of a second enzyme. J Bacteriol 170:2121–2125
Cleton-Jansen AM, Goosen N, Odle G, Van de Putte P (1988b) Nucleotide sequence of the gene coding for quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus. Nucleic Acids Res 16:6228
Cleton-Jansen AM, Goosen N, Vink K, Van de Putte P (1989) Cloning, characterization and DNA sequencing of the gene encoding the Mr 50,000 quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus. Mol Gen Genet 217:430–436
Cleton-Jansen AM, Goosen N, Fayet O, van de Putte P (1990) Cloning, mapping, and sequencing of the gene encoding Escherichia coli quinoprotein glucose dehydrogenase. J Bacteriol 172:6308–6315
Chomczynski P, Qasba PK (1984) Alkaline transfer of DNA to plastic membrane. Biochem Biophys Res Commun 122:340–344
Creaser EH, Murali C, Britt KA (1990) Protein engineering of alcohol dehydrogenases: effects of amino acid changes at positions 93 and 48 of yeast ADH1. Prot Engng 3:523–526
De Ley J (1961) Comparative carboxyhydrate metabolism and a proposal for phylogenetic relationship of the acetic acid bacteria. J Gen Microbiol 24:31–50
Deveraux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:333–446
Ditta G, Stanfield S, Corbin D, Helenski DR (1980) Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351
Duine JA, Frank J, Van Zeeland K (1979) Glucose dehydrogenase from Acinetobacter calcoaceticus, a “quinoprotein”. FEBS Lett 108:443–446
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Goosen N, Vermaas DAM, Van de Putte P (1987) Cloning of the genes involved in synthesis of coenzyme pyrrolo-quinoline quinone from Acinetobacter calcoaceticus. J Bacteriol 169:303–307
Hauge JG (1966) Glucose dehydrogenase: Pseudomonas sp. and Bacterium antitratum. Methods Enzymol 9:92–98
Hommes RJW, Postma PW, Neijssel OM, Tempest DW, Dokter P, Duine JA (1984) Evidence of a quinoprotein glucose dehydrogenase apoenzyme in several strains of Escherichia coli. FEMS Microbiol Lett 24:329–333
Karn J, Brenner S, Barnett L, Cesareni G (1980) Novel bacteriophage I cloning vector. Proc Natl Acad Sci USA 77:5172
Kunkel TA, Roberts JD, Zakour RA (1987) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492
Matsushita K, Shinagawa E, Inoue T, Adachi O, Ameyama M (1986) Immunological evidence for two types of PQQ dependent d-glucose dehydrogenase in bacterial membranes and the location of the enzyme in Escherichia coli. FEMS Microbiol Lett 37:141–144
Messing J, Crea R, Seeburg PH (1981) A system for shotgun DNA sequencing. Nucleic Acids Res 9:309–321
Pronk JT, Levering PR, Olijve W, Van Dijken JP (1989) Role of NADP-dependent and quinoprotein glucose dehydrogenases in gluconic acid production by Gluconobacter oxydans. Enzyme Microb Technol 11:160–164
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Simon R, Priefer U, Puhler A (1983) A broad host range mobilization system for in vitro engineering: transposon mutagenesis in gram-negative bacteria. Bio/Technology 1:742–791
Tabor S, Richardson CC (1987) DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci USA 84:4767–4771
Van Rijn PA, Goosen N, Turk SCHJ, Van de Putte P (1989) Regulation of phage Mu repressor transcription by IHF depends on the level of early transcription. Nucleic Acids Res 17:10203–10211
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequence of M13 mp18 and pUC19 vectors. Gene 55:103–119
Zoller MJ, Smith M (1987) Oligonucleotide-directed mutagenesis: A simple method using two oligonucleotide primers and single stranded DNA template. Methods Enzymol 154:329
Author information
Authors and Affiliations
Additional information
Communicated by C. van den Hondel
Rights and permissions
About this article
Cite this article
Cleton-Jansen, AM., Dekker, S., van de Putte, P. et al. A single amino acid substitution changes the substrate specificity of quinoprotein glucose dehydrogenase in Gluconobacter oxydans . Molec. Gen. Genet. 229, 206–212 (1991). https://doi.org/10.1007/BF00272157
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00272157