Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs

Abstract Aerobic methanotrophic bacteria utilize methane as a growth substrate but are unable to grow on any sugars. In this study we have shown that two obligate methanotrophs, Methylotuvimicrobium alcaliphilum 20Z and Methylobacter luteus IMV-B-3098, possess functional glucose dehydrogenase (GDH)...

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Autores principales:	Olga N. Rozova, Galina A. Ekimova, Nikolai V. Molochkov, Alexander S. Reshetnikov, Valentina N. Khmelenina, Ildar I. Mustakhimov
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Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d7de1e9c2d7b40bea5a76ded68314eaa
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spelling	oai:doaj.org-article:d7de1e9c2d7b40bea5a76ded68314eaa2021-12-02T16:45:11ZEnzymes of an alternative pathway of glucose metabolism in obligate methanotrophs10.1038/s41598-021-88202-x2045-2322https://doaj.org/article/d7de1e9c2d7b40bea5a76ded68314eaa2021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-88202-xhttps://doaj.org/toc/2045-2322Abstract Aerobic methanotrophic bacteria utilize methane as a growth substrate but are unable to grow on any sugars. In this study we have shown that two obligate methanotrophs, Methylotuvimicrobium alcaliphilum 20Z and Methylobacter luteus IMV-B-3098, possess functional glucose dehydrogenase (GDH) and gluconate kinase (GntK). The recombinant GDHs from both methanotrophs were homotetrameric and strongly specific for glucose preferring NAD+ over NADP+. GDH from Mtm. alcaliphilum was most active at pH 10 (V max = 95 U/mg protein) and demonstrated very high K m for glucose (91.8 ± 3.8 mM). GDH from Mb. luteus was most active at pH 8.5 (V max = 43 U/mg protein) and had lower K m for glucose (16 ± 0.6 mM). The cells of two Mtm. alcaliphilum double mutants with deletions either of the genes encoding GDH and glucokinase (gdh ─/glk ─) or of the genes encoding gluconate kinase and glucokinase (gntk ─/glk ─) had the lower glycogen level and the higher contents of intracellular glucose and trehalose compared to the wild type strain. The gntk ─/glk ─ knockout mutant additionally accumulated gluconic acid. These data, along with bioinformatics analysis, demonstrate that glycogen derived free glucose can enter the Entner–Doudoroff pathway or the pentose phosphate cycle in methanotrophs, bypassing glycolysis via the gluconate shunt.Olga N. RozovaGalina A. EkimovaNikolai V. MolochkovAlexander S. ReshetnikovValentina N. KhmeleninaIldar I. MustakhimovNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Olga N. Rozova Galina A. Ekimova Nikolai V. Molochkov Alexander S. Reshetnikov Valentina N. Khmelenina Ildar I. Mustakhimov Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs
description	Abstract Aerobic methanotrophic bacteria utilize methane as a growth substrate but are unable to grow on any sugars. In this study we have shown that two obligate methanotrophs, Methylotuvimicrobium alcaliphilum 20Z and Methylobacter luteus IMV-B-3098, possess functional glucose dehydrogenase (GDH) and gluconate kinase (GntK). The recombinant GDHs from both methanotrophs were homotetrameric and strongly specific for glucose preferring NAD+ over NADP+. GDH from Mtm. alcaliphilum was most active at pH 10 (V max = 95 U/mg protein) and demonstrated very high K m for glucose (91.8 ± 3.8 mM). GDH from Mb. luteus was most active at pH 8.5 (V max = 43 U/mg protein) and had lower K m for glucose (16 ± 0.6 mM). The cells of two Mtm. alcaliphilum double mutants with deletions either of the genes encoding GDH and glucokinase (gdh ─/glk ─) or of the genes encoding gluconate kinase and glucokinase (gntk ─/glk ─) had the lower glycogen level and the higher contents of intracellular glucose and trehalose compared to the wild type strain. The gntk ─/glk ─ knockout mutant additionally accumulated gluconic acid. These data, along with bioinformatics analysis, demonstrate that glycogen derived free glucose can enter the Entner–Doudoroff pathway or the pentose phosphate cycle in methanotrophs, bypassing glycolysis via the gluconate shunt.
format	article
author	Olga N. Rozova Galina A. Ekimova Nikolai V. Molochkov Alexander S. Reshetnikov Valentina N. Khmelenina Ildar I. Mustakhimov
author_facet	Olga N. Rozova Galina A. Ekimova Nikolai V. Molochkov Alexander S. Reshetnikov Valentina N. Khmelenina Ildar I. Mustakhimov
author_sort	Olga N. Rozova
title	Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs
title_short	Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs
title_full	Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs
title_fullStr	Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs
title_full_unstemmed	Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs
title_sort	enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/d7de1e9c2d7b40bea5a76ded68314eaa
work_keys_str_mv	AT olganrozova enzymesofanalternativepathwayofglucosemetabolisminobligatemethanotrophs AT galinaaekimova enzymesofanalternativepathwayofglucosemetabolisminobligatemethanotrophs AT nikolaivmolochkov enzymesofanalternativepathwayofglucosemetabolisminobligatemethanotrophs AT alexandersreshetnikov enzymesofanalternativepathwayofglucosemetabolisminobligatemethanotrophs AT valentinankhmelenina enzymesofanalternativepathwayofglucosemetabolisminobligatemethanotrophs AT ildarimustakhimov enzymesofanalternativepathwayofglucosemetabolisminobligatemethanotrophs
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Enzymes of an alternative pathway of glucose metabolism in obligate methanotrophs

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