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纤细裸藻中乙醇酸的两个碳原子在转化为丝氨酸过程中的不同代谢命运。

Different metabolic fate of two carbons of glycolate in its conversion to serine in Euglena gracilis z.

作者信息

Yokota A, Komura H, Kitaoka S

出版信息

Arch Biochem Biophys. 1985 Nov 1;242(2):498-506. doi: 10.1016/0003-9861(85)90236-x.

Abstract

In our preceding work (A. Yokota, Y. Nakano, and S. Kitaoka, 1978, Agric. Biol. Chem. 42, 121-129), extensive decarboxylation of glycolate carboxyl carbon during its metabolism in Euglena gracilis suggested occurrence of a metabolic pathway of glycolate different from that of higher C3 plants. In the present report, we establish the Euglena glycolate pathway from characteristics of the decarboxylation of the carboxyl carbon and from the metabolic fate of hydroxymethyl carbon of glycolate. The ratio of the decarboxylation of the carboxyl carbon of glycolate to the total metabolized carbon increased with increasing metabolic rate in an asymptotic fashion. Thus, the ratio was 20% at the metabolic rate of 0.05 nmol of glycolate/10(6) cells/min, but it was over 60% at the rate of more than 0.35 nmol/10(6) cells/min after 2 min of incubation. Metabolic products were also changed depending on the rate of metabolism of glycolate; glycine was the main product at the low rate of glycolate metabolism and the contribution of glycine was reversed by the increased contribution of evolved CO2 at the high rates. At the metabolic rate of 1.5 nmol of glycolate/10(6) cells/min, the rate of the decarboxylation was 1.0 nmol of CO2/10(6) cells/min, which could not be explained by the extremely low activity of glycine synthase in Euglena. Experiments with [2-14C]glycolate showed that exogenously added formate and methionine caused accumulation of radioactive formate. Based on these results, we have proposed that the glycolate metabolism of E. gracilis consists of glycine and formate pathways and that the relative contribution of both pathways to the glycolate metabolism depends on the metabolic rate of glycolate.

摘要

在我们之前的工作中(横田明、中野洋、北冈幸雄,1978年,《农业与生物化学》42卷,121 - 129页),纤细裸藻在乙醇酸代谢过程中,乙醇酸羧基碳发生大量脱羧,这表明存在一条与高等C3植物不同的乙醇酸代谢途径。在本报告中,我们根据羧基碳的脱羧特性以及乙醇酸羟甲基碳的代谢归宿,确立了裸藻的乙醇酸途径。乙醇酸羧基碳的脱羧量与总代谢碳量的比值随着代谢速率的增加以渐近方式上升。因此,在乙醇酸代谢速率为0.05 nmol/10⁶个细胞/分钟时,该比值为20%,但在孵育2分钟后,当代谢速率超过0.35 nmol/10⁶个细胞/分钟时,该比值超过60%。代谢产物也会根据乙醇酸的代谢速率而变化;在乙醇酸代谢速率较低时,甘氨酸是主要产物,而在高代谢速率下,随着释放的二氧化碳贡献增加,甘氨酸的贡献则相反。在乙醇酸代谢速率为1.5 nmol/10⁶个细胞/分钟时,脱羧速率为1.0 nmol二氧化碳/10⁶个细胞/分钟,这无法用裸藻中极低的甘氨酸合酶活性来解释。用[2 - ¹⁴C]乙醇酸进行的实验表明,外源添加的甲酸和甲硫氨酸会导致放射性甲酸积累。基于这些结果,我们提出纤细裸藻的乙醇酸代谢由甘氨酸途径和甲酸途径组成,并且这两条途径对乙醇酸代谢的相对贡献取决于乙醇酸的代谢速率。

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