Purification and properties of invertase from the flowers of Woodfordia fruticosa.
Invertase (beta-fructofuranosidase, EC 3.2.1.26) was purified from the flowers of Woodfordia fruticosa, which is used to prepare certain fermented Ayurvedic drugs. The enzyme was purified to near homogeneity as judged by native PAGE with a yield of 10.7%, using (NH4)2SO4 fractionation, followed by gel filtration through Sepharose 4B and DEAE cellulose chromatography at pH 6.8 and 4.42. The molecular mass of the purified enzyme as determined by elution through Sepharose 4B gel column was found to be approximately 280 kDa. SDS-PAGE of the purified enzyme showed that the enzyme is composed of three subunits with molecular mass of 66, 43 and 40 kDa. The enzyme showed a broad pH optimum between 4.0-7.0. Optimum assay temperature was 37 degrees C and above 45 degrees C, the enzyme activity slowly declined and inactivated around 80 degrees C. The apparent Km value of the enzyme for sucrose was 160 mM.
Department of Chemistry, University of Kelaniya, Sri Lanka. bandu@kln.ac.lk
M K B
MK
Weersaooriya
H P
HP
Yatawara
D002482:Cellulose / Q000737:chemistry; D002845:Chromatography / Q000379:methods; D002850:Chromatography, Gel; D004798:Enzymes / Q000737:chemistry; D035264:Flowers / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D012685:Sepharose / Q000737:chemistry; D013696:Temperature; D013997:Time Factors; D031565:Woodfordia / Q000201:enzymology*; D043324:beta-Fructofuranosidase / Q000096:biosynthesis*
D004798:Enzymes; D002482:Cellulose; D012685:Sepharose; D043324:beta-Fructofuranosidase