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

Mohammad R. Haque, Aiko Hirowatari, Nonoko Nai, Shigeki Furuya, Kohji Yamamoto

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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.

Original languageEnglish
Article numbere21594
JournalArchives of insect biochemistry and physiology
Volume102
Issue number2
DOIs
Publication statusPublished - Oct 1 2019

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Glycine Hydroxymethyltransferase
glycine hydroxymethyltransferase
Bombyx
Bombyx mori
silkworms
NADP
NADP (coenzyme)
serine
Serine
Amino Acids
Mutagenesis
glycine (amino acid)
Metabolism
mutagenesis
Glycine
Escherichia coli
Carbon
amino acid sequences
Complementary DNA
Amino Acid Sequence

All Science Journal Classification (ASJC) codes

  • Physiology
  • Biochemistry
  • Insect Science

Cite this

Serine hydroxymethyltransferase from the silkworm Bombyx mori : Identification, distribution, and biochemical characterization. / Haque, Mohammad R.; Hirowatari, Aiko; Nai, Nonoko; Furuya, Shigeki; Yamamoto, Kohji.

In: Archives of insect biochemistry and physiology, Vol. 102, No. 2, e21594, 01.10.2019.

Research output: Contribution to journalArticle

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