Concanavalin A Immobilized Affinity Adsorbents for Reversible Use in Yeast Invertase Adsorption

Abstract

Concanavalin A (Con A) immobilized poly(2-hydroxyethyl methacrylate) (PHEMA) beads were investigated for specific adsorption of yeast invertase from aqueous solutions. PHEMA beads were prepared by a suspension polymerization technique with an average size of 150-200 microm, and activated by epichlorohydrin. Con A was then immobilized by covalent binding onto these beads. The maximum Con A immobilization was found to be 10 mg/g. The invertase-loading capability of the PHEMA/Con A beads was 107 mg/g. The maximum invertase adsorption capacity on the PHEMA/Con A adsorbents was observed at pH 5.0. The values of the Michaelis constant K(m) of invertase were significantly larger upon adsorption, indicating decreased affinity by the enzyme for its substrate, whereas V(max) was smaller for the adsorbed invertase. Adsorption improved the pH stability of the enzyme as well as its temperature stability. Thermal stability was found to increase with adsorption. The adsorbed enzyme activity was found to be quite stable in repeated experiments. Storage stability of adsorbed invertase.

Concanavalin A immobilized poly(ethylene glycol dimethacrylate) based affinity cryogel matrix and usability of invertase immobilization

“…The maximum invertase adsorption capacity of these beads observed as 35.2 mg/g polymer. Yavuz et al [5] produced Concanavalin A attached poly(HEMA) beads and investigated its invertase adsorption capacity. Maximum invertase loading capability of the Poly(HEMA)/Con A beads was found to be 107 mg/g polymer.…”

“…Affinity purification of glycoenzymes and glycopeptides on immobilized lectins is widely employed for preparative purposes [1][2][3][4][5]. Con A, a 104 kDa tetrameric protein, is one of the most extensively investigated lectins due to its ability to form specific complexes with ␣-d-mannose and ␣-d-glucose containing glycoconjugates.…”

Adsorption of lysozyme from aqueous solutions by a novel bentonite–tyrptophane (Bent–Trp) microcomposite affinity sorbent

“…The method also offers the reusability of expensive supports after inactivation of immobilized enzyme. As the main problem with polymer supports is lower pH stabilities and thermal stabilities, many supports have been unemployed to immobilize enzymes, the most important being polymers [3][4][5]. Inorganic supports provide better thermal stabilities and hence there is immense scope for research in this area [6].…”

Orthogonal Transformations on Solid Substrates: Efficient Avenues to Surface Modification

“…[141][142][143][144] because of their large specific surface area. However, the modification of microspheres is inevitably required to increase their cell compatibility and bioactivity.…”