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Title: ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs

Abstract

The 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 generamore » 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.« less

Authors:
 [1];  [2];  [1];  [3];  [3];  [4]
  1. Russian Academy of Sciences (RAS), Pushchino (Russian Federation). Federal Research Center. Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences. G.K. Skryabin Inst. of Biochemistry and Physiology of Microorganisms. Lab. of Methylotrophy
  2. Pushchino State Inst. of Natural Sciences, Pushchino (Russian Federation). Dept. of Microbiology and Biotechnology
  3. Russian Academy of Sciences (RAS), Pushchino (Russian Federation). Federal Research Center. Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences. G.K. Skryabin Inst. of Biochemistry and Physiology of Microorganisms; Pushchino State Inst. of Natural Sciences, Pushchino (Russian Federation). Dept. of Microbiology and Biotechnology
  4. Russian Academy of Sciences (RAS), Pushchino (Russian Federation). Federal Research Center. Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences. G.K. Skryabin Inst. of Biochemistry and Physiology of Microorganisms
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division
OSTI Identifier:
1628467
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Microorganisms
Additional Journal Information:
Journal Volume: 7; Journal Issue: 2; Journal ID: ISSN 2076-2607
Publisher:
MDPI
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Microbiology; ATP-glucokinase; Methylomonas sp. 12; methanotrophs; polyphosphate-glucokinase

Citation Formats

Reshetnikov, Alexander, Solntseva, Natalia, Rozova, Olga, Mustakhimov, Ildar, Trotsenko, Yuri, and Khmelenina, Valentina. ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs. United States: N. p., 2019. Web. doi:10.3390/microorganisms7020052.
Reshetnikov, Alexander, Solntseva, Natalia, Rozova, Olga, Mustakhimov, Ildar, Trotsenko, Yuri, & Khmelenina, Valentina. ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs. United States. doi:10.3390/microorganisms7020052.
Reshetnikov, Alexander, Solntseva, Natalia, Rozova, Olga, Mustakhimov, Ildar, Trotsenko, Yuri, and Khmelenina, Valentina. Fri . "ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs". United States. doi:10.3390/microorganisms7020052. https://www.osti.gov/servlets/purl/1628467.
@article{osti_1628467,
title = {ATP- and Polyphosphate-Dependent Glucokinases from Aerobic Methanotrophs},
author = {Reshetnikov, Alexander and Solntseva, Natalia and Rozova, Olga and Mustakhimov, Ildar and Trotsenko, Yuri and Khmelenina, Valentina},
abstractNote = {The 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.},
doi = {10.3390/microorganisms7020052},
journal = {Microorganisms},
number = 2,
volume = 7,
place = {United States},
year = {2019},
month = {2}
}

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