Hydrogel for immobilization of one or more enzyme(s) and method for preparing the same

A method of preparing a biocompatible hydrogel,

• comprising the following steps:
• a) Providing a first polysaccharide and a second polysaccharide,
• optionally wherein the first polysaccharide and/or the second polysaccharide being independently from each other selected from the group consisting of pullulan, alginate, cellulose, hyaluronan, dextran, lichenin, lentinan and mixtures thereof,
• with n being the number of the monomeric repeating unit of the first and/or second polysaccharide and with n being an integer from 10 to 10000,
• b) optionally carboxymethylation of at least one OH-group of the first and/or second polysaccharide;
• c) functionalization of the first polysaccharide with one or more linker unit(s) of the structure -A-X, when the monomeric repeating unit of the first polysaccharide does not comprise a carboxylic acid residue, or functionalization of the first polysaccharide with one or more linker unit(s) of the structure —X, when the monomeric repeating unit of the first polysaccharide comprises a carboxylic acid residue,
• wherein A is —(CH2)d—C(O)— or —C(O)—NH—, wherein d is an integer from 1 to 3,
• wherein X is selected from the group consisting of —NH—(CH2)rN3, —NH—(CH2CH2O)s—CH2CH2N3, and —NH—(CH2—CH2—C(O))t—CH2—CH2—N3,
• with r being an integer from 2 to 20,
• s being an integer from 1 to 15, and
• t being an integer from 1 to 15;
• and
• functionalization of the second polysaccharide with one or more linker unit(s) of the structure -A′-Y, when the monomeric repeating unit of the second polysaccharide does not comprise a carboxylic acid residue, or functionalization of the second polysaccharide with one or more linker unit(s) of the structure —Y, when the monomeric repeating unit of the second polysaccharide comprises a carboxylic acid residue,
• wherein A′ is —(CH2)d—C(O)— or —C(O)—NH—, wherein d is an integer from 1 to 3,
• wherein Y is selected from the group consisting of —NH—(CH2)r-Q, —NH—(CH2CH2O)s—CH2CH2Q and —NH—(CH2—CH2—C(O))t—CH2—CH2-Q,
• wherein Q is
• wherein M, M′=H or Me, and
• wherein W=OMe, OEt, OH, NH2 or NHMe,
• d) optionally having the addition of one or more enzyme(s) to the mixture formed by steps a)-c), and
• e) incubation of the mixture formed by steps a)-d) in an aqueous medium at a temperature being in one of the range from 25° C. to 70° C., or at 40° C., for one of at least 1 hour, or for 1 to 10 hours.