Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase

Abstract A unicellular cyanobacterium Synechocystis sp. PCC 6803 possesses a unique tricarboxylic acid (TCA) cycle, wherein the intracellular citrate levels are approximately 1.5–10 times higher than the levels of other TCA cycle metabolite. Aconitase catalyses the reversible isomerisation of citrat...

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Autores principales:	Maki Nishii, Shoki Ito, Noriaki Katayama, Takashi Osanai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f9888978176f4e71834adfe9f1845d2d
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spelling	oai:doaj.org-article:f9888978176f4e71834adfe9f1845d2d2021-12-02T18:53:08ZBiochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase10.1038/s41598-021-96432-22045-2322https://doaj.org/article/f9888978176f4e71834adfe9f1845d2d2021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96432-2https://doaj.org/toc/2045-2322Abstract A unicellular cyanobacterium Synechocystis sp. PCC 6803 possesses a unique tricarboxylic acid (TCA) cycle, wherein the intracellular citrate levels are approximately 1.5–10 times higher than the levels of other TCA cycle metabolite. Aconitase catalyses the reversible isomerisation of citrate and isocitrate. Herein, we biochemically analysed Synechocystis sp. PCC 6803 aconitase (SyAcnB), using citrate and isocitrate as the substrates. We observed that the activity of SyAcnB for citrate was highest at pH 7.7 and 45 °C and for isocitrate at pH 8.0 and 53 °C. The K m value of SyAcnB for citrate was higher than that for isocitrate under the same conditions. The K m value of SyAcnB for isocitrate was 3.6-fold higher than the reported K m values of isocitrate dehydrogenase for isocitrate. Therefore, we suggest that citrate accumulation depends on the enzyme kinetics of SyAcnB, and 2-oxoglutarate production depends on the chemical equilibrium in this cyanobacterium.Maki NishiiShoki ItoNoriaki KatayamaTakashi OsanaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maki Nishii Shoki Ito Noriaki Katayama Takashi Osanai Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase
description	Abstract A unicellular cyanobacterium Synechocystis sp. PCC 6803 possesses a unique tricarboxylic acid (TCA) cycle, wherein the intracellular citrate levels are approximately 1.5–10 times higher than the levels of other TCA cycle metabolite. Aconitase catalyses the reversible isomerisation of citrate and isocitrate. Herein, we biochemically analysed Synechocystis sp. PCC 6803 aconitase (SyAcnB), using citrate and isocitrate as the substrates. We observed that the activity of SyAcnB for citrate was highest at pH 7.7 and 45 °C and for isocitrate at pH 8.0 and 53 °C. The K m value of SyAcnB for citrate was higher than that for isocitrate under the same conditions. The K m value of SyAcnB for isocitrate was 3.6-fold higher than the reported K m values of isocitrate dehydrogenase for isocitrate. Therefore, we suggest that citrate accumulation depends on the enzyme kinetics of SyAcnB, and 2-oxoglutarate production depends on the chemical equilibrium in this cyanobacterium.
format	article
author	Maki Nishii Shoki Ito Noriaki Katayama Takashi Osanai
author_facet	Maki Nishii Shoki Ito Noriaki Katayama Takashi Osanai
author_sort	Maki Nishii
title	Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase
title_short	Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase
title_full	Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase
title_fullStr	Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase
title_full_unstemmed	Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase
title_sort	biochemical elucidation of citrate accumulation in synechocystis sp. pcc 6803 via kinetic analysis of aconitase
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/f9888978176f4e71834adfe9f1845d2d
work_keys_str_mv	AT makinishii biochemicalelucidationofcitrateaccumulationinsynechocystissppcc6803viakineticanalysisofaconitase AT shokiito biochemicalelucidationofcitrateaccumulationinsynechocystissppcc6803viakineticanalysisofaconitase AT noriakikatayama biochemicalelucidationofcitrateaccumulationinsynechocystissppcc6803viakineticanalysisofaconitase AT takashiosanai biochemicalelucidationofcitrateaccumulationinsynechocystissppcc6803viakineticanalysisofaconitase
_version_	1718377363757072384

Biochemical elucidation of citrate accumulation in Synechocystis sp. PCC 6803 via kinetic analysis of aconitase

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