J. Bacteriol., Jan 1996, 149-155, Vol 178, No. 1
Copyright © 1996, American Society for Microbiology

Characterization and phylogeny of the pfp gene of Amycolatopsis methanolica encoding PPi-dependent phosphofructokinase

AM Alves, WG Meijer, JW Vrijbloed and L Dijkhuizen
Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Haren, The Netherlands.

The actinomycete Amycolatopsis methanolica employs a PPi-dependent phosphofructokinase (PPi-PFK) (EC 2.7.1.90) with biochemical characteristics similar to those of both ATP- and PPi-dependent enzymes during growth on glucose. A 2.3-kb PvuII fragment hybridizing to two oligonucleotides based on the amino-terminal amino acid sequence of PPi- PFK was isolated from a genomic library of A. methanolica. Nucleotide sequence analysis of this fragment revealed the presence of an open reading frame encoding a protein of 340 amino acids with a high degree of similarity to PFK proteins. Heterologous expression of this open reading frame in Escherichia coli gave rise to a unique 45-kDa protein displaying a high level of PPi-PFK activity. The open reading frame was therefore designated pfp, encoding the PPi-PFK of A. methanolica. Upstream and transcribed divergently from pfp, a partial open reading frame (aroA) similar to 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase-encoding genes was identified. The partial open reading frame (chiA) downstream from pfp was similar to chitinase genes from Streptomyces species. A phylogenetic analysis of the ATP- and PPi- dependent proteins showed that PPi-PFK enzymes are monophyletic, suggesting that the two types of PFK evolved from a common ancestor.

This article has been cited by other articles:

• Kloosterman, H., Hessels, G. I., Vrijbloed, J. W., Euverink, G. J., Dijkhuizen, L. (2003). (De)regulation of key enzyme steps in the shikimate pathway and phenylalanine-specific pathway of the actinomycete Amycolatopsis methanolica. Microbiology 149: 3321-3330 [Abstract] [Full Text]  
• Alves, A. M. C. R., Euverink, G. J. W., Santos, H., Dijkhuizen, L. (2001). Different Physiological Roles of ATP- and PPi-Dependent Phosphofructokinase Isoenzymes in the Methylotrophic Actinomycete Amycolatopsis methanolica. J. Bacteriol. 183: 7231-7240 [Abstract] [Full Text]  
• Gosset, G., Bonner, C. A., Jensen, R. A. (2001). Microbial Origin of Plant-Type 2-Keto-3-Deoxy-D-arabino-Heptulosonate 7-Phosphate Synthases, Exemplified by the Chorismate- and Tryptophan-Regulated Enzyme from Xanthomonas campestris. J. Bacteriol. 183: 4061-4070 [Abstract] [Full Text]  
• Siebers, B., Klenk, H.-P., Hensel, R. (1998). PPi-Dependent Phosphofructokinase from Thermoproteus tenax, an Archaeal Descendant of an Ancient Line in Phosphofructokinase Evolution. J. Bacteriol. 180: 2137-2143 [Abstract] [Full Text]  
• Matheron, C., Delort, A.-M., Gaudet, G., Forano, E., Liptaj, T. (1998). 13C and 1H Nuclear Magnetic Resonance Study of Glycogen Futile Cycling in Strains of the Genus Fibrobacter. Appl. Environ. Microbiol. 64: 74-81 [Abstract] [Full Text]