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

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

Research output: Contribution to journalArticle

9 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 May 1

Keywords

  • Aryl sulfotransferase
  • Bombyx mori
  • Gallic acid
  • Insect

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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