Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Methylococcus capsulatus Bath

Reshetnikov, Alexander S., Rozova, Olga N., Khmelenina, Valentina N., Mustakhimov, Ildar I., Beschastny, Alexander P., Murrell, J. Colin and Trotsenko, Yuri A. (2008) Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Methylococcus capsulatus Bath. FEMS Microbiology Letters, 288 (2). pp. 202-210. ISSN 1574-6968

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Abstract

An active pyrophosphate-dependent 6-phosphofructokinase (PPi-PFK) from the thermotolerant methanotroph Methylococcus capsulatus Bath, containing a six-residue polyhistidine tag, was characterized. The enzyme was homodimeric (2 × 45 kDa), nonallosteric and most active at pH 7.0. PPi-PFK catalyzed reactions of PPi-dependent phosphorylation of fructose-6-phosphate (F-6-P) (Km 2.27 mM and Vmax 7.6 U mg-1 of protein), sedoheptulose-7-phosphate (Km 0.027 mM and Vmax 31 U mg-1) and ribulose-5-phosphate. In the reaction with F-6-P, the apparent Km for PPi was 0.027 mM, while in the reverse reaction, Km for orthophosphate was 8.69 mM and that for fructose-1,6-bisphosphate 0.328 mM (Vmax 9.0 U mg-1). Phylogenetically, M. capsulatus PPi-PFK was most similar to PPi-PFKs from the lithoautotrophic ammonia oxidizers Nitrosomonas europaea (74.0%), Nitrosospira multiformis (73.6%) and Betaproteobacterial methylotroph Methylibium petroleiphilum PM1 (71.6% identity). Genes coding PPi-PFK and a putative V-type H+-translocating pyrophosphatase (H+-PPi-ase) were cotranscribed as an operon. The potential significance of the PPi-PFK for regulation of carbon and energy fluxes in M. capsulatus Bath is discussed.

Item Type:	Article
Faculty \ School:	Faculty of Science > School of Environmental Sciences
UEA Research Groups:	Faculty of Science > Research Centres > Centre for Ecology, Evolution and Conservation Faculty of Science > Research Groups > Environmental Biology Faculty of Science > Research Groups > Resources, Sustainability and Governance (former - to 2018) Faculty of Science > Research Groups > Marine and Atmospheric Sciences (former - to 2017)
Related URLs:	http://dx.doi.org/10.1111/j.1574-6968.20...
Depositing User:	Rhiannon Harvey
Date Deposited:	27 Mar 2012 16:14
Last Modified:	24 Oct 2022 01:03
URI:	https://ueaeprints.uea.ac.uk/id/eprint/38528
DOI:	10.1111/j.1574-6968.2008.01366.x

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