Serine hydroxymethyltransferase from the silkworm Bombyx mori: Identification, distribution, and biochemical characterization.
Serine hydroxymethyltransferase from the silkworm Bombyx mori: Identification, distribution, and biochemical characterization.
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家蚕丝氨酸羟甲基转移酶：鉴定、分布和生化特征。
DOI：
10.1002/arch.21594
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发表时间：
2019
影响因子：
2.2
通讯作者：
Kohji Yamamoto
中科院分区：
文献类型：
--
作者：
M. Haque;A. Hirowatari;N. Nai;S. Furuya;Kohji Yamamoto
丝氨酸羟甲基转移酶 (SHMT) 催化丝氨酸和四氢叶酸 (THF) 相互转化为甘氨酸和亚甲基四氢叶酸。克隆并测序了编码家蚕 SHMT (bmSHMT) 的 cDNA。推导的氨基酸序列由 465 个氨基酸组成，并被发现与其他 SHMT 具有同源性。重组 bmSHMT 在大肠杆菌中过表达并纯化至均质。该酶在 pH 3.0 和 30°C 时表现出最佳活性，并且在酸性条件下稳定。在烟酰胺腺嘌呤二核苷酸磷酸 (NADP+) 存在下，THF 的 Km 和 kcat /Km 值分别为 0.055 mM 和 0.081 mM-1 s-1 ，而针对 NADP+ 的 Km 和 kcat /Km 值分别为 0.16 mM 和 0.018 mM-1 s-1 ，针对 L-丝氨酸的 Km 和 kcat /Km 值为分别为1.8 mM和0.0022 mM-1 s-1。诱变实验表明 His119、His132 和 His135 对于酶活性很重要。我们的结果提供了对家蚕一碳代谢的作用和调节机制的深入了解。
Serine hydroxymethyltransferase (SHMT) catalyzes the interconversion of serine and tetrahydrofolate (THF) to glycine and methylenetetrahydrofolate. cDNA encoding Bombyx mori SHMT (bmSHMT) was cloned and sequenced. The deduced amino acid sequence consisted of 465 amino acids and was found to share homology with other SHMTs. Recombinant bmSHMT was overexpressed in Escherichia coli and purified to homogeneity. The enzyme showed optimum activity at pH 3.0 and 30°C and was stable under acidic conditions. The Km and kcat /Km values for THF in the presence of Nicotinamide adenine dinucleotide phosphate (NADP+ ) were 0.055 mM and 0.081 mM-1 s-1 , respectively, whereas those toward NADP+ were 0.16 mM and 0.018 mM-1 s-1 and toward l-serine were 1.8 mM and 0.0022 mM-1 s-1 , respectively. Mutagenesis experiments revealed that His119, His132, and His135 are important for enzymatic activity. Our results provide insight into the roles and regulation mechanism of one-carbon metabolism in the silkworm B. mori.
甲酸乙酸梭菌 5,10-次甲基四氢叶酸环化水解酶的纯化和性质。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Clark,JE;Ljungdahl,LG
通讯作者：
Ljungdahl,LG
影响因子：
3.5
作者：
Girgis, S;Nasrallah, IM;Stover, PJ
通讯作者：
Stover, PJ
影响因子：
2.9
作者：
Szebenyi, DME;Musayev, FN;Schirch, V
通讯作者：
Schirch, V