Details

Autor(en) / Beteiligte

• Yamamoto, Kohji
• Zhang, Pingbo
• Miake, Fumio
• Kashige, Nobuhiro
• Aso, Yoichi
• Banno, Yutaka
• Fujii, Hiroshi

Titel

Cloning, expression and characterization of theta-class glutathione S-transferase from the silkworm, Bombyx mori

Ist Teil von

• Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 2005-07, Vol.141 (3), p.340-346

Ort / Verlag

England: Elsevier Inc

Erscheinungsjahr

2005

Quelle

MEDLINE

Beschreibungen/Notizen

• This study focused on glutathione S-transferase (GST), one of the detoxification enzymes, from the silkworm, Bombyx mori (GSTT1). A cDNA encoding a putative GST was amplified by reverse transcriptase-polymerase chain reaction and sequenced. The deduced amino acid sequence revealed 59%, 57% and 56% identities to theta-class GSTs of Musca domestica, Anopheles gambiae and Drosophila melanogaster, respectively. GSTT1 was also estimated to be close to those GSTs in a phylogenetic tree. Recombinant GST (rGSTT1) was functionally overexpressed in Escherichia coli in a soluble form, purified to homogeneity, and characterized. The pH-optimum of rGSTT1 was broad from pH 4 to 9 and rGSTT1 retained more than 75% of its original activity after incubation at pH 5–11. Incubation for 30 min at temperatures below 50 °C also affected the activity insignificantly. The Michaelis constant for 1-chloro-2,4-dinitrobenzene was 0.48 mM.