Biochemical Properties and Phylogeny of Hydroxypyruvate Reductases from Methanotrophic Bacteria with Different C₁-Assimilation Pathways

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S. Y. But^1*, S. V. Egorova², V. N. Khmelenina¹, and Y. A. Trotsenko^1,2*

¹Skryabin Institute of Biochemistry and Physiology of Microorganisms, Laboratory of Methylotrophy, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia; E-mail: flash20063@rambler.ru, trotsenko@ibpm.pushchino.ru

²Pushchino State Institute of Natural Sciences, 142290 Pushchino, Moscow Region, Russia

^* To whom correspondence should be addressed.

Received July 11, 2017; Revision received August 22, 2017

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In the aerobic methanotrophic bacteria Methylomicrobium alcaliphilum 20Z, Methylococcus capsulatus Bath, and Methylosinus trichosporium OB3b, the biochemical properties of hydroxypyruvate reductase (Hpr), an indicator enzyme of the serine pathway for assimilation of reduced C₁-compounds, were comparatively analyzed. The recombinant Hpr obtained by cloning and heterologous expression of the hpr gene in Escherichia coli catalyzed NAD(P)H-dependent reduction of hydroxypyruvate or glyoxylate, but did not catalyze the reverse reactions of D-glycerate or glycolate oxidation. The absence of the glycerate dehydrogenase activity in the methanotrophic Hpr confirmed a key role of the enzyme in utilization of C₁-compounds via the serine cycle. The enzyme from Ms. trichosporium OB3b realizing the serine cycle as a sole assimilation pathway had much higher special activity and affinity in comparison to Hpr from Mm. alcaliphilum 20Z and Mc. capsulatus Bath assimilating carbon predominantly via the ribulose monophosphate (RuMP) cycle. The hpr gene was found as part of gene clusters coding the serine cycle enzymes in all sequenced methanotrophic genomes except the representatives of the Verrucomicrobia phylum. Phylogenetic analyses revealed two types of Hpr: (i) Hpr of methanotrophs belonging to the Gammaproteobacteria class, which use the serine cycle along with the RuMP cycle, as well as of non-methylotrophic bacteria belonging to the Alphaproteobacteria class; (ii) Hpr of methylotrophs from Alpha- and Betaproteobacteria classes that use only the serine cycle and of non-methylotrophic representatives of Betaproteobacteria. The putative role and origin of hydroxypyruvate reductase in methanotrophs are discussed.

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KEY WORDS: hydroxypyruvate reductase, methanotrophs, serine cycle of carbon assimilation

DOI: 10.1134/S0006297917110074

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