Immobilized biocatalysts

Claim 1

A method for immobilization of microbial cells comprising: (a) forming a solution of an agar or carrageenan hydrogel; (b) introducing microbial cells into the hydrogel solution, said microbial cells being at temperature 40°-48° C. when agar is used, and 37°-50° C. when ccarrageenan is used; (c) forming spherical gel beads of the microbial cell containing hydrogel solution, such beads created by contacting the hydrogel solution with a water-immiscible liquid at a temperature below that of the hydrogel solution; (d) activating the microbial cells by incubating the spherical beads containing culture for 12-50 hours, such incubation resulting in the formation of colonies of microbial cells in the spherical beads which produce CO 2 thereby decreasing resistance of the beads to diffusion; (e) diffusing the monomer, acrylamide (ACAM), the cross-linking agent, N,N'-methylene-bis-acrylamide (BIS), and the accelerator, β-dimethylaminopropionitrile (DMAPN) or N,N,N',N'-tetramethylenediamine (TEMED) into the beads by introducing the ACAM, BIS, and DMAPN or TEMED to the gel beads in solution; (f) after diffusion removing the solution of ACAM, BIS and either DMAPN or TEMED; (g) contacting the gel beads with a polymerization initiator selected from the group consisting of K 2 S 2 O 8 and (NH 4 ) 2 S 2 O 8 to cause polymerization; (h) sparging the solution with nitrogen during polymerization; (i) stirring the beads so as to increase uptake of polymerization initiator, and; (j) washing the polymerized beads with water to remove unpolymerized molecules.

Claim 2

A method for immobilization of microbial cells comprising: (a) forming a solution of an agar or carrageenan hydrogel; (b) introducing microbial cells into the hydrogel solution, said microbial cells being at temperature 40°-48° C. when agar is used, and 37°-50° C. when ccarrageenan is used; (c) forming spherical gel beads of the microbial cell containing hydrogel solution, such beads created by contacting the hydrogel solution with a water-immiscible liquid at a temperature below that of the hydrogel solution; (d) activating the microbial cells by incubating the spherical beads containing culture for 12-50 hours, such incubation resulting in the formation of colonies of microbial cells in the spherical beads which produce CO 2 thereby decreasing resistance of the beads to diffusion; (e) diffusing the monomer, acrylamide (ACAM), the cross-linking agent, N,N'-methylene-bis-acrylamide (BIS), and the accelerator, β-dimethylaminopropionitrile (DMAPN) or N,N,N',N'-tetramethylenediamine (TEMED) into the beads by introducing the ACAM, BIS, and DMAPN or TEMED to the gel beads in solution; (f) after diffusion removing the solution of ACAM, BIS and either DMAPN or TEMED; (g) contacting the gel beads with a polymerization initiator selected from the group consisting of K 2 S 2 O 8 and (NH 4 ) 2 S 2 O 8 to cause polymerization; (h) sparging the solution with nitrogen during polymerization; (i) stirring the beads so as to increase uptake of polymerization initiator, and; (j) washing the polymerized beads with water to remove unpolymerized molecules.