Journal of Agricultural Science and Technology
Journal of Agricultural Science and Technology
JAST
Agriculture
http://jast.modares.ac.ir
1
admin
1680-7073
2345-3737
10.22034/jast
en
jalali
1393
8
1
gregorian
2014
11
1
16
6
online
1
fulltext
en
Purification and Characterization of Midgut α-Glucosidase from Larvae of the Rice Green Caterpillar, Naranga aenescens Moore
Application of chemical pesticides has increased significantly worldwide and has raised serious concerns about environmental pollutions. One of the encouraging trends to minimize pesticide risk is production of resistant plants containing toxic proteins against insect pests. Considering the importance of purification and characterization of digestive enzymes in the production of resistant plants, in this study an <em>α</em>-glucosidase from the <em>Naranga aenescens </em>Moore's midgut was purified by ammonium sulfate precipitation, ion exchange chromatography on DEAE-sepharose, and concentrating through ultrafiltration. The apparent molecular mass of the enzyme was 48 kDa determined by SDS-PAGE. The optimum pH and temperature of the enzyme were 6.0 and 45°C, respectively. The irreversible thermoinactivation of the enzyme showed that it was highly stable at 35ºC but moderately stable at 40 and 45ºC. Zn<sup>2+</sup>, Hg<sup>2+</sup>, Co<sup>2+</sup> at 10 and 20 mM, and Ba<sup>+2</sup>only in 20 mM strongly inhibited the <em>α</em>-glucosidase activity. Ba<sup>2+</sup> and Ca<sup>2+</sup> only at 10 mM, EDTA and Hg<sub>2</sub><sup>2+</sup> only at 20 mM and Mg<sup>2+</sup> at 10 and 20 mM significantly increased the enzyme activity. The <em>K<sub>m</sub></em> and <em>K<sub>cat</sub> </em>values for the <em>α</em>-glucosidase were 0.54 mM and 3.62 min<sup>-1</sup>, respectively, when <em>p</em>-Nitrophenyl-α-D-glucopyranoside (pNαG) was used as a substrate.
Ion exchange chromatography,Enzyme activity,Kinetic parameters,Thermostability Kinetic parameters,Toxic proteins
1253
1266
http://jast.modares.ac.ir/browse.php?a_code=A-23-1000-3323&slc_lang=en&sid=23
N
Memarizadeh
N
Memarizadeh
100319475328460054072
100319475328460054072
No
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Islamic Republic of Iran.
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Islamic Republic of Iran.
P.
Zamani
P.
Zamani
100319475328460054071
100319475328460054071
No
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Islamic Republic of Iran.
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Islamic Republic of Iran.
R. H.
Sajedi
R. H.
Sajedi
100319475328460054083
100319475328460054083
No
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Islamic Republic of Iran.
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Islamic Republic of Iran.
M.
Ghadamyari
M.
Ghadamyari
100319475328460054070
100319475328460054070
Yes
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Islamic Republic of Iran.
Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Islamic Republic of Iran.