Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Methylococcus capsulatus Bath
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 0378-1097Full text not available from this repository.
Official URL: http://dx.doi.org/10.1111/j.1574-6968.2008.01366.x
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.|
|Number of Pages:||9|
|Page Range:||pp. 202-210|
|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 18.104.22.16871 (MAS)|
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