Purification and characterization of β-glucosidase from greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae).
The greater wax moth, Galleria mellonella, is one of the most ruinous pests of honeycomb in the world. Beta-glucosidases are a type of digestive enzymes that hydrolytically catalyzes the beta-glycosidic linkage of glycosides. Characterization of the beta-glucosidase in G. mellonella could be a significant stage for a better comprehending of its role and establishing a safe and effective control procedure primarily against G. mellonella and also some other insect pests. Laboratory reared final instar stage larvae were randomly selected and homogenized for beta-glucosidase activity assay and subsequent analysis. The enzyme was purified to apparent homogeneity by salting out with ammonium sulfate and using sepharose-4B-l-tyrosine-1-naphthylamine hydrophobic interaction chromatography. The purification was 58-fold with an overall enzyme yield of 29%. The molecular mass of the protein was estimated as ca. 42 kDa. The purified beta-glucosidase was effectively active on para/ortho-nitrophenyl-beta-d-glucopyranosides (p-/o-NPG) with Km values of 0.37 and 1.9 mM and Vmax values of 625 and 189 U/mg, respectively. It also exhibits different levels of activity against para-nitrophenyl-β-d-fucopyranoside (p-NPF), para/ortho-nitrophenyl β-d-galactopyranosides (p-/o-NPGal) and p-nitrophenyl 1-thio-β-d-glucopyranoside. The enzyme was competitively inhibited by beta-gluconolactone and also was very tolerant to glucose against p-NPG as substrate. The Ki and IC50 values of δ-gluconolactone were determined as 0.021 and 0.08 mM while the enzyme was more tolerant to glucose inhibition with IC50 value of 213.13 mM for p-NPG.
Department of Basic Sciences/Biochemistry, Faculty of Veterinary, Balikesir University, Balıkesir, Turkey.
Hatibe Ertürk
HE
Kara
Yusuf
Y
Turan
Aylin
A
Er
Mesut
M
Acar
Sabiha
S
Tümay
Selma
S
Sinan
D000818:Animals; D001516:Bees / Q000469:parasitology*; D002847:Chromatography, Agarose; D004591:Electrophoresis, Polyacrylamide Gel; D005936:Glucans / Q000378:metabolism; D005942:Gluconates / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D020128:Inhibitory Concentration 50; D007700:Kinetics; D007783:Lactones / Q000494:pharmacology; D007814:Larva / Q000201:enzymology; D009036:Moths / Q000201:enzymology* / Q000502:physiology; D013696:Temperature; D001617:beta-Glucosidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*
D005936:Glucans; D005942:Gluconates; D007783:Lactones; D001617:beta-Glucosidase; D005947:Glucose; C010730:beta-glucono-1,5-lactone
Galleria mellonella; characterization; purification; β-glucosidase