کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9954170 | 1540195 | 2018 | 39 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Low molecular weight α-galactosidase from black gram (Vigna mungo): Purification and insights towards biochemical and biophysical properties
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
A hitherto unknown low molecular weight form of α-galactosidase (VM-αGal-P) from germinating black gram (Vigna mungo) seeds was purified (324â¯U/mg specific activity, 1157-fold purification, ~45â¯kDa) using ion-exchange (DEAE-cellulose, CM-sepharose), gel filtration (Sephadex G-75) and affinity (Con-A Sepharose 4B) chromatography but with poor yield (0.75%). Partially purified enzyme (VM-αGal) (146.3â¯U/mg specific activity, 522.5-fold purification) was used for further studies. VM-αGal showed optimal activity at pHâ¯5 and 55â¯Â°C. Hg2+ and SDS completely inhibited VM-αGal activity. The Km, Vmax and catalytic efficiency (kcat/Km) of VM-αGal for pNPG and raffinose was 0.99, 17.23â¯mM, 1.66, 0.146â¯Î¼molâ¯mlâ1â¯minâ1, and 0.413, 0.0026â¯sâ1â¯mMâ1, respectively. VM-αGal was competitively inhibited by galactose (Ki 7.70â¯mM). Thermodynamic parameters [activation enthalpy (ÎH), activation entropy (ÎS) and free energy (ÎG)] of VM-αGal at 45-51â¯Â°C showed that VM-αGal was in a less energetic state and had susceptibility towards denaturation. Temperature-induced structural unfolding studies of VM-αGal probed by fluorescence, and far-UV CD spectroscopy revealed significant loss in tertiary structure and a steep decline in β-sheet content at 45-65â¯Â°C, and above 55â¯Â°C, respectively. VM-αGal improved the nutritional quality of soymilk by hydrolyzing raffinose family oligosaccharides (26.5% and 18.45% decrease in stachyose and raffinose, respectively).
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Biological Macromolecules - Volume 119, November 2018, Pages 770-778
Journal: International Journal of Biological Macromolecules - Volume 119, November 2018, Pages 770-778
نویسندگان
Ramadevi Mutra, Juby Elsa Joseph, Deepesh Panwar, Gaurav Singh Kaira, Mukesh Kapoor,