Reshetnikov, Alexander S., Rozova, Olga N., Khmelenina, V. N. (Valentina Nikolaevna), Mustakhimov, Ildar I., Beschastny, Alexander P., Murrell, J. C. (J. Colin) and Trotsenko, Yuri A.. (2008) Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Methylococcus capsulatus Bath. FEMS Microbiology Letters, Volume 288 (Number 2). pp. 202-210. ISSN 0378-1097

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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 x 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) (K-m 2.27 mM and V-max 7.6 U mg(-1) of protein), sedoheptulose-7-phosphate (K-m 0.027 mM and V-max 31 U mg(-1)) and ribulose-5-phosphate. In the reaction with F-6-P, the apparent K-m for PPi was 0.027 mM, while in the reverse reaction, K-m for orthophosphate was 8.69 mM and that for fructose-1,6-bisphosphate 0.328 mM (V-max 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:	Journal Article
Subjects:	Q Science > QR Microbiology
Divisions:	Faculty of Science > Life Sciences (2010- ) > Biological Sciences ( -2010)
Library of Congress Subject Headings (LCSH):	Methanotrophs -- Research
Journal or Publication Title:	FEMS Microbiology Letters
Publisher:	Wiley-Blackwell Publishing Ltd.
ISSN:	0378-1097
Official Date:	November 2008
Dates:	Date Event November 2008 Published
Volume:	Volume 288
Number:	Number 2
Number of Pages:	9
Page Range:	pp. 202-210
Identification Number:	10.1111/j.1574-6968.2008.01366.x
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	Rossiĭskiĭ fond fundamentalʹnykh issledovaniĭ [Russian Foundation for Basic Research] (RFBR), Ministry of Education and Science of the Russian Federation
Grant number:	08-04-01484a (RFBR), RF RNP 2.1.1.2671 (MAS)
URI:	http://wrap.warwick.ac.uk/id/eprint/29251

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