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J Bacteriol. Apr 1986; 166(1): 319–327.
PMCID: PMC214595

Expression of the Escherichia coli pfkA gene in Alcaligenes eutrophus and in other gram-negative bacteria.

Abstract

The Escherichia coli pfkA gene has been cloned in the non-self-transmissible vector pVK101 from hybrid plasmids obtained from the Clarke and Carbon clone bank, resulting in the plasmids pAS300 and pAS100; the latter plasmid also encoded the E. coli tpi gene. These plasmids were transferred by conjugation to mutants of Alcaligenes eutrophus which are unable to grow on fructose and gluconate due to lack of 2-keto-3-deoxy-6-phosphogluconate aldolase activity. These transconjugants recovered the ability to grow on fructose and harbored pAS100 or pAS300. After growth on fructose, the transconjugants contained phosphofructokinase at specific activities between 0.73 and 1.83 U/mg of protein, indicating that the E. coli pfkA gene is readily expressed in A. eutrophus and that the utilization of fructose occurs via the Embden-Meyerhof pathway instead of the Entner-Doudoroff pathway. In contrast, transconjugants of the wild type of A. eutrophus, which are potentially able to catabolize fructose via both pathways, grew at a decreased rate on fructose and during growth on fructose did not stably maintain pAS100 or pAS300. Indications for a glycolytic futile cycling of fructose 6-phosphate and fructose 1,6-bisphosphate are discussed. Plasmid pA 100 was also transferred to 14 different species of gram-negative bacteria. The pfkA gene was expressed in most of these species. In addition, most transconjugants of these strains and of A. eutrophus exhibited higher specific activities of triosephosphate isomerase than did the corresponding parent strains.

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Selected References

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  • Abdelal AT, Schlegel HG. Purification and regulatory properties of fructose 1,6-diphosphatase from Hydrogenomonas eutropha. J Bacteriol. 1974 Oct;120(1):304–310. [PMC free article] [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Blackkolb F, Schlegel HG. Katabolische Repression und Enzymhemmung durch molekularen Wasserstoff bei Hydrogenomonas. Arch Mikrobiol. 1968;62(2):129–143. [PubMed]
  • Bowien B, Schlegel HG. Isolierung und Charakterisierung katabolischer Defektmutanten von Hydrogenomonas eutropha Stamm H 16. II. Mutanten mit einem Defekt in der 2-Keto-3-desoxy-6-phosphogluconat-Aldolase. Arch Mikrobiol. 1972;87(3):221–234. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Clarke L, Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. [PubMed]
  • Clewell DB, Helinski DR. Properties of a supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry. 1970 Oct 27;9(22):4428–4440. [PubMed]
  • Colman A, Byers MJ, Primrose SB, Lyons A. Rapid purification of plasmid DNAs by hydroxyapatite chromatography. Eur J Biochem. 1978 Nov 2;91(1):303–310. [PubMed]
  • Don RH, Pemberton JM. Properties of six pesticide degradation plasmids isolated from Alcaligenes paradoxus and Alcaligenes eutrophus. J Bacteriol. 1981 Feb;145(2):681–686. [PMC free article] [PubMed]
  • Figurski DH, Meyer RJ, Helinski DR. Suppression of Co1E1 replication properties by the Inc P-1 plasmid RK2 in hybrid plasmids constructed in vitro. J Mol Biol. 1979 Sep 25;133(3):295–318. [PubMed]
  • Fraenkel DG, Kotlarz D, Buc H. Two fructose 6-phosphate kinase activities in Escherichia coli. J Biol Chem. 1973 Jul 10;248(13):4865–4866. [PubMed]
  • Friedrich B, Hogrefe C, Schlegel HG. Naturally occurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus. J Bacteriol. 1981 Jul;147(1):198–205. [PMC free article] [PubMed]
  • GOTTSCHALK G, EBERHARDT U, SCHLEGEL HG. VERWERTUNG VON FRUCTOSE DURCH HYDROGENOMONAS H 16. (I.) Arch Mikrobiol. 1964 Apr 2;48:95–108. [PubMed]
  • Hellinga HW, Evans PR. Nucleotide sequence and high-level expression of the major Escherichia coli phosphofructokinase. Eur J Biochem. 1985 Jun 3;149(2):363–373. [PubMed]
  • Knauf VC, Nester EW. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. [PubMed]
  • Kuhn M, Steinbüchel A, Schlegel HG. Hydrogen evolution by strictly aerobic hydrogen bacteria under anaerobic conditions. J Bacteriol. 1984 Aug;159(2):633–639. [PMC free article] [PubMed]
  • Mergeay M, Houba C, Gerits J. Extrachromosomal inheritance controlling resistance to cadmium, cobalt, copper and zinc ions: evidence from curing in a Pseudomonas [proceedings]. Arch Int Physiol Biochim. 1978 May;86(2):440–442. [PubMed]
  • Neidhardt FC, Vaughn V, Phillips TA, Bloch PL. Gene-protein index of Escherichia coli K-12. Microbiol Rev. 1983 Jun;47(2):231–284. [PMC free article] [PubMed]
  • Pfitzner J, Schlegel HG. Denitrifikation bei Hydrogenomonas eutropha Stamm H 16. Arch Mikrobiol. 1973;90(3):199–211. [PubMed]
  • QUAYLE JR, KEECH DB. Carbon dioxide and formate utilization by formate-grown Pseudomonas oxalaticus. Biochim Biophys Acta. 1958 Jul;29(1):223–225. [PubMed]
  • Reutz I, Schobert P, Bowien B. Effect of phosphoglycerate mutase deficiency on heterotrophic and autotrophic carbon metabolism of Alcaligenes eutrophus. J Bacteriol. 1982 Jul;151(1):8–14. [PMC free article] [PubMed]
  • SCHLEGEL HG, KALTWASSER H, GOTTSCHALK G. [A submersion method for culture of hydrogen-oxidizing bacteria: growth physiological studies]. Arch Mikrobiol. 1961;38:209–222. [PubMed]
  • Senior PJ, Dawes EA. The regulation of poly-beta-hydroxybutyrate metabolism in Azotobacter beijerinckii. Biochem J. 1973 May;134(1):225–238. [PMC free article] [PubMed]
  • Shimosaka M, Fukuda Y, Murata K, Kimura A. Application of hybrid plasmids carrying glycolysis genes to ATP production by Escherichia coli. J Bacteriol. 1982 Oct;152(1):98–103. [PMC free article] [PubMed]
  • Silverman PM. Gene cpxA is a new addition to the linkage map of Escherichia coli K-12. J Bacteriol. 1982 Apr;150(1):425–428. [PMC free article] [PubMed]
  • Steinbüchel A, Schlegel HG. NAD-linked L(+)-lactate dehydrogenase from the strict aerobe Alcaligenes eutrophus. 1. Purification and properties. Eur J Biochem. 1983 Feb 1;130(2):321–328. [PubMed]
  • Steinbüchel A, Schlegel HG. NAD-linked L(+)-lactate dehydrogenase from the strict aerobe alcaligenes eutrophus. 2. Kinetic properties and inhibition by oxaloacetate. Eur J Biochem. 1983 Feb 1;130(2):329–334. [PubMed]
  • Thomson JA. E. coli phosphofructokinase synthesized in vitro from a ColE1 hybrid plasmid. Gene. 1977 Jul;1(5-6):347–356. [PubMed]
  • Thomson J, Gerstenberger PD, Goldberg DE, Gociar E, Orozco de Silva A, Fraenkel DG. ColE1 hybrid plasmids for Escherichia coli genes of glycolysis and the hexose monophosphate shunt. J Bacteriol. 1979 Jan;137(1):502–506. [PMC free article] [PubMed]
  • WALDE E. [Studies on growth and synthesis of stored substance by Hydrogenomonas]. Arch Mikrobiol. 1962;43:109–137. [PubMed]

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