Characterization of recombinant PPi-dependent 6-phosphofructokinases from Methylosinus trichosporium OB3b and Methylobacterium nodulans ORS 2060

• Published in: Biochemistry (Moscow) Vol. 77; no. 3; pp. 288 - 295
• Main Authors: Rozova, O. N., Khmelenina, V. N., Trotsenko, Y. A.
• Format: Journal Article
• Language: English
• Published: Dordrecht SP MAIK Nauka/Interperiodica 01.03.2012; Springer; Springer Nature B.V
• Subjects: Adenosine diphosphate; Amino Acid Sequence; Amino acids; Bacteria; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Chemical properties; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - metabolism; Enzymes; Fructose; Fructosephosphates - metabolism; Kinetics; Life Sciences; Methylobacterium; Methylobacterium - chemistry; Methylobacterium - classification; Methylobacterium - enzymology; Methylobacterium - genetics; Methylosinus trichosporium; Methylosinus trichosporium - chemistry; Methylosinus trichosporium - classification; Methylosinus trichosporium - enzymology; Methylosinus trichosporium - genetics; Microbiology; Molecular Sequence Data; Phosphates; Phosphofructokinase-1 - chemistry; Phosphofructokinase-1 - genetics; Phosphofructokinase-1 - metabolism; Phosphorylation; Phylogeny; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; sedoheptulose-1,7-bisphosphatase; methylotrophic bacteria; pyrophosphate-dependent 6-phosphofructokinase; gluconeogenesis; glycolysis
• ISSN: 0006-2979; 1608-3040
• DOI: 10.1134/S0006297912030078

The properties of the purified recombinant PPi-dependent 6-phosphofructokinases (PPi-PFKs) from the methanotroph Methylosinus trichosporium OB3b and rhizospheric phytosymbiont Methylobacterium nodulans ORS 2060 were determined. The dependence of activities of PPi-PFK-His 6 -tag from Ms. trichosporium OB3b (6 × 45 kDa) and PPi-PFK from Mb. nodulans ORS 2060 (4 × 43 kDa) on the concentrations of substrates of forward and reverse reactions conformed to Michaelis-Menten kinetics. Besides fructose-6-phosphate, the enzymes also phosphorylated sedoheptulose-7-phosphate. ADP or AMP (1 mM each) inhibited activity of the Ms. trichosporium PPi-PFK but did not affect the activity of the Mb. nodulans enzyme. Preference of PPi-PFKs to fructose-1,6-bisphosphate implied a predominant function of the enzymes in hexose phosphate synthesis in these bacteria. PPi-PFKs from the methylotrophs have low similarity of translated amino acid sequences (17% identity) and belong to different phylogenetic subgroups of type II 6-phosphofructokinases. The relationship of PPi-PFKs with microaerophilic character of Ms. trichosporium OB3b and adaptation of Mb. nodulans ORS 2060 to anaerobic phase of phytosymbiosis are discussed.