Cloning and expression of a novel sulfotransferase with unique substrate specificity from Bombyx mori

Kenji Hattori, Mio Hirayama, Hiroko Suzuki, Hiroshi Hamamoto, Kazuhisa Sekimizu, Hiroomi Tamura

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We identified a cDNA encoding a putative cytosolic sulfotransferase (SULT) by searching the expressed sequence tag database of Bombyx mori, and subsequently obtained the full-length cDNA for this gene via rapid amplification of cDNA ends (RACE). We designated this gene bmST1, and showed by sequence analysis that it belongs to a novel SULT family. The tissue specificity of bmST1 mRNA expression was examined in fifth instar larvae by reverse transcriptase-polymerase chain reaction (RT-PCR), and transcripts were detectable in the silk gland, gut, fat body, and Malpighian tube. A recombinant form of bmST1 was then expressed using a gluthathione S-transferase (GST) gene fusion system, and it was purified from Escherichia coli. Purified bmST1 did not exhibit sulfating activity toward SULT substrates such as 4-nitrophenol, vanillin, hydroxysteroids, or monoamines. Surprisingly, however, recombinant bmST1 showed considerable activity toward 4-nitrocatechol and also gallate esters, although the catechins are not sulfated by this enzyme.

Original languageEnglish
Pages (from-to)1044-1051
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume71
Issue number4
DOIs
Publication statusPublished - 2007

Fingerprint

sulfotransferases
Sulfotransferases
Bombyx
Cloning
Bombyx mori
substrate specificity
Substrate Specificity
Organism Cloning
molecular cloning
Complementary DNA
Genes
Substrates
Hydroxysteroids
silk glands
p-nitrophenol
monoamines
Organ Specificity
Fat Body
Silk
rapid amplification of cDNA ends

Keywords

  • Aryl sulfotransferase
  • Bombyx mori
  • Gallic acid
  • Insect

ASJC Scopus subject areas

  • Bioengineering
  • Biotechnology
  • Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Food Science

Cite this

Cloning and expression of a novel sulfotransferase with unique substrate specificity from Bombyx mori. / Hattori, Kenji; Hirayama, Mio; Suzuki, Hiroko; Hamamoto, Hiroshi; Sekimizu, Kazuhisa; Tamura, Hiroomi.

In: Bioscience, Biotechnology and Biochemistry, Vol. 71, No. 4, 2007, p. 1044-1051.

Research output: Contribution to journalArticle

Hattori, Kenji ; Hirayama, Mio ; Suzuki, Hiroko ; Hamamoto, Hiroshi ; Sekimizu, Kazuhisa ; Tamura, Hiroomi. / Cloning and expression of a novel sulfotransferase with unique substrate specificity from Bombyx mori. In: Bioscience, Biotechnology and Biochemistry. 2007 ; Vol. 71, No. 4. pp. 1044-1051.
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