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

被引:10
作者
Nishii, Maki [1 ]
Ito, Shoki [1 ]
Katayama, Noriaki [1 ]
Osanai, Takashi [1 ]
机构
[1] Meiji Univ, Sch Agr, Tama Ku, 1-1-1 Higashimita, Kawasaki, Kanagawa 2148571, Japan
基金
日本科学技术振兴机构;
关键词
TRICARBOXYLIC-ACID CYCLE; RNA-BINDING; CLEAVAGE REACTION; SIGMA-FACTOR; ENZYME; IRON; METABOLITES; ACNB; MITOCHONDRIAL; PURIFICATION;
D O I
10.1038/s41598-021-96432-2
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 degrees C and for isocitrate at pH 8.0 and 53 degrees 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.
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页数:12
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