ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs
AbstractThe genes encoding adenosine triphosphate (ATP)- and polyphosphate (polyP)-dependent glucokinases (Glk) were identified in the aerobic obligate methanotroph Methylomonas sp. 12. The recombinant proteins were obtained by the heterologous expression of the glk genes in Esherichia coli. ATP-Glk behaved as a multimeric protein consisting of di-, tri-, tetra-, penta- and hexamers with a subunit molecular mass of 35.5 kDa. ATP-Glk phosphorylated glucose and glucosamine using ATP (100% activity), uridine triphosphate (UTP) (85%) or guanosine triphosphate (GTP) (71%) as a phosphoryl donor and exhibited the highest activity in the presence of 5 mM Mg2+ at pH 7.5 and 65 °C but was fully inactivated after a short-term incubation at this temperature. According to a gel filtration in the presence of polyP, the polyP-dependent Glk was a dimeric protein (2 × 28 kDa). PolyP-Glk phosphorylated glucose, mannose, 2-deoxy-D-glucose, glucosamine and N-acetylglucosamine using polyP as the phosphoryl donor but not using nucleoside triphosphates. The Km values of ATP-Glk for glucose and ATP were about 78 μM, and the Km values of polyP-Glk for glucose and polyP(n=45) were 450 and 21 μM, respectively. The genomic analysis of methanotrophs showed that ATP-dependent glucokinase is present in all sequenced methanotrophs, with the exception of the genera Methylosinus and Methylocystis, whereas polyP-Glks were found in all species of the genus Methylomonas and in Methylomarinum vadi only. This work presents the first characterization of polyphosphate specific glucokinase in a methanotrophic bacterium. View Full-Text
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Reshetnikov, A.S.; Solntseva, N.P.; Rozova, O.N.; Mustakhimov, I.I.; Trotsenko, Y.A.; Khmelenina, V.N. ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs. Microorganisms 2019, 7, 52.
Reshetnikov AS, Solntseva NP, Rozova ON, Mustakhimov II, Trotsenko YA, Khmelenina VN. ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs. Microorganisms. 2019; 7(2):52.Chicago/Turabian Style
Reshetnikov, Alexander S.; Solntseva, Natalia P.; Rozova, Olga N.; Mustakhimov, Ildar I.; Trotsenko, Yuri A.; Khmelenina, Valentina N. 2019. "ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs." Microorganisms 7, no. 2: 52.
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