Expression and characterization of a recombinant Drosophila tyramine-β-hydroxylase in silkworm infected with recombinant baculovirus

Ahmed M.H. Ali, Man Lee, Mayumi Yoshida, Kosuke Sakashita, Jumpei Torii, Takahiro Kusakabe, Akinori Hirashima

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The insect nervous system contains biogenic amines such octopamine (OA), which is synthesized from tyramine (TA) by catalysis of tyramine-β-hydroxylase (TβH). In this study, the Drosophila 70 kDa tyramine-β-hydroxylase (DmTβH) protein was purified after the recombinant nucleopolyhedrovirus isolated from Bombyx mori (BmNPV) containing the TβH gene was injected into the hemocoel of the fifth instar larvae from the d17 B. mori strain. Western blot analysis revealed an immunoreactive band with a molecular mass of 70 kDa. The products formed by incubating the enzyme reaction mixture were separated and detected by reverse phase high-performance liquid chromatography. The optimum pH, temperature, and incubation time for the conversion of TA to OA were 7.6, 25 °C, and 30 min, respectively. The inhibitory experiments using various concentrations of 1-(2-methoxy-5-methylphenyl) imidazole-2(3. H)-thione (MMIT) showed that MMIT inhibited DmTβH dose-dependently and that this method can be applied for screening DmTβH inhibitors.

Original languageEnglish
Pages (from-to)567-572
Number of pages6
JournalJournal of Asia-Pacific Entomology
Volume15
Issue number4
DOIs
Publication statusPublished - Dec 1 2012

Fingerprint

tyramine
Baculoviridae
silkworms
Drosophila
octopamine
Bombyx mori
Bombyx mori nucleopolyhedrovirus
hemocoel
imidazoles
reversed-phase liquid chromatography
biogenic amines
enzymatic reactions
catalytic activity
nervous system
Western blotting
instars
high performance liquid chromatography
molecular weight
screening
insects

All Science Journal Classification (ASJC) codes

  • Insect Science

Cite this

Expression and characterization of a recombinant Drosophila tyramine-β-hydroxylase in silkworm infected with recombinant baculovirus. / Ali, Ahmed M.H.; Lee, Man; Yoshida, Mayumi; Sakashita, Kosuke; Torii, Jumpei; Kusakabe, Takahiro; Hirashima, Akinori.

In: Journal of Asia-Pacific Entomology, Vol. 15, No. 4, 01.12.2012, p. 567-572.

Research output: Contribution to journalArticle

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