Purification of alpha-glucosidae and invertase from bakers' yeast on modified polymeric supports.
In the present work Amberlite XAD-16 and Indion NPA-1, Polystyrene Divinylbenzene macroreticular spherical resins, have been evaluated quantitatively as supports for the adsorption and isolation of the yeast proteins and the enzymes, invertase and alpha-glucosidase. Modification of these supports has been carried out by surface grafting using acrylate polymers to reduce the hydrophobicity and nonspecific adsorption of proteins. Good grafting efficiency, in excess of 90%, has been obtained using ultrasonic irradiation for the surface activation of polystyrene resins. XAD-16 has higher adsorption capacities for the total yeast proteins as well as for both the enzymes, alpha-glucosidase and invertase, than NPA-1 in its respective native and grafted form. Adsorption capacities of XAD-16 and NPA-1 in their respective native and grafted forms for alpha-glucosidase are higher than the capacities for invertase. Nonspecific adsorption of total proteins has been reduced considerably after the grafting of acrylate polymers on hydrophobic supports. At the same time selectivity for the adsorption of both the enzymes has been enhanced on grafted supports. The overall solid-liquid adsorption mass transfer coefficient values (Kla) estimated for adsorption of invertase on XAD are lower than those for alpha-glucosidase. Native and grafted resins could be regenerated and reused for adsorption of alpha-glucosidase for two regeneration cycles studied. Storage stability of invertase and alpha-glucosidase is the same on native and grafted form of XAD-16 and is more than the enzymes in the free form.
Lothe
R R
RR
Chemical Engineering Division, University of Mumbai, Matunga, India.
Purohit
S S
SS
Shaikh
S S
SS
Malshe
V C
VC
Pandit
A B
AB
D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification*; D007475:Ion Exchange Resins / Q000737:chemistry*; D011108:Polymers; D011137:Polystyrenes / Q000737:chemistry*; D011145:Polyvinyls / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013055:Spectrophotometry, Infrared; D014867:Water / Q000737:chemistry; D000520:alpha-Glucosidases / Q000737:chemistry / Q000302:isolation & purification*; D043324:beta-Fructofuranosidase
D005656:Fungal Proteins; D007475:Ion Exchange Resins; D011108:Polymers; D011137:Polystyrenes; D011145:Polyvinyls; D014867:Water; C074337:Amberlite XAD-16; C012101:XAD-4 resin; D006026:Glycoside Hydrolases; D000520:alpha-Glucosidases; D043324:beta-Fructofuranosidase