Application of porous microspheres prepared by SPG (Shirasu porous glass) emulsification as immobilizing carriers of glucoamylase (GluA)

✍ Scribed by Shinzo Omi; Kazuyoshi Kaneko; Akira Nakayama; Ken'ichi Katami; Tetsuya Taguchi; Mamoru Iso; Masatoshi Nagai; Guang-Hui Ma

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 240 KB
• Volume: 65
• Category: Article
• ISSN: 0021-8995
• DOI: 10.1002/(sici)1097-4628(19970926)65:13<2655::aid-app7>3.0.co;2-a

✦ Synopsis

Fairly uniform spheres of crosslinked polystyrene (PS) and polymethyl methacrylate (PMMA), prepared by a particular emulsification process using SPG (Shirasu Porous Glass) membranes and subsequent suspension polymerization, were applied for immobilizing carriers of Glucoamylase (GluA). A mixture of monomers, solvents, and oil-soluble initiator was allowed to permeate through the micropores of SPG, suspended in an aqueous solution of poly(vinyl alcohol), and polymerized while retaining the narrow size distribution during polymerization. A small amount of acrylic acid or glycidyl methacrylate (GMA) was incorporated for the immobilization of GluA via covalent bonding. Although GluA has been regarded as being difficult to retain its activity after the immobilization process, a porous structure of the carriers definitely favored the immobilization, and a maximum 55% relative activity (RA) was obtained by the physical adsorption to PMMA spheres. The reaction of epoxide in GMA with 6aminocaproic acid provided a spacer arm for the carboxyl group. An improvement of activity was expected by the incorporation of the spacer arms; however, barely noticeable activity was observed for PMMA carriers either by the physical adsorption or by the covalent bonding. A slight improvement was observed for PS carriers with spacers compared to the carriers without them. The diffusion process of oligosaccharides in the porous carriers seemed to retard the rate of hydrolysis in the case of largest carriers, 60 mm PS-DVB-AA spheres. The activity of immobilized GluA was retained during a long storage period of more than 150 days, some of them even increasing gradually, while the activity of native GluA dropped to zero after 100 days.