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Serine hydroxymethyltransferase from the silkworm Bombyx mori: Identification, distribution, and biochemical characterization.

Abstract

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.

Authors+Show Affiliations

Department of Bioscience and Biotechnology, Kyushu University Graduate School, Nishi-ku, Fukuoka, Japan.Department of Bioscience and Biotechnology, Kyushu University Graduate School, Nishi-ku, Fukuoka, Japan.Department of Bioscience and Biotechnology, Kyushu University Graduate School, Nishi-ku, Fukuoka, Japan.Department of Bioscience and Biotechnology, Kyushu University Graduate School, Nishi-ku, Fukuoka, Japan.Department of Bioscience and Biotechnology, Kyushu University Graduate School, Nishi-ku, Fukuoka, Japan.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31298425

Citation

Haque, Mohammad R., et al. "Serine Hydroxymethyltransferase From the Silkworm Bombyx Mori: Identification, Distribution, and Biochemical Characterization." Archives of Insect Biochemistry and Physiology, 2019, pp. e21594.
Haque MR, Hirowatari A, Nai N, et al. Serine hydroxymethyltransferase from the silkworm Bombyx mori: Identification, distribution, and biochemical characterization. Arch Insect Biochem Physiol. 2019.
Haque, M. R., Hirowatari, A., Nai, N., Furuya, S., & Yamamoto, K. (2019). Serine hydroxymethyltransferase from the silkworm Bombyx mori: Identification, distribution, and biochemical characterization. Archives of Insect Biochemistry and Physiology, pp. e21594. doi:10.1002/arch.21594.
Haque MR, et al. Serine Hydroxymethyltransferase From the Silkworm Bombyx Mori: Identification, Distribution, and Biochemical Characterization. Arch Insect Biochem Physiol. 2019 Jul 12;e21594. PubMed PMID: 31298425.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Serine hydroxymethyltransferase from the silkworm Bombyx mori: Identification, distribution, and biochemical characterization. AU - Haque,Mohammad R, AU - Hirowatari,Aiko, AU - Nai,Nonoko, AU - Furuya,Shigeki, AU - Yamamoto,Kohji, Y1 - 2019/07/12/ PY - 2019/7/13/entrez KW - biosynthesis KW - cobalamin KW - one-carbon metabolism KW - site-directed mutagenesis KW - tetrahydrofolate SP - e21594 EP - e21594 JF - Archives of insect biochemistry and physiology JO - Arch. Insect Biochem. Physiol. N2 - 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. SN - 1520-6327 UR - https://www.unboundmedicine.com/medline/citation/31298425/Serine_hydroxymethyltransferase_from_the_silkworm_Bombyx_mori:_Identification,_distribution,_and_biochemical_characterization L2 - https://doi.org/10.1002/arch.21594 DB - PRIME DP - Unbound Medicine ER -