Protein modification to provide enzyme activity

Chemistry: molecular biology and microbiology – Carrier-bound or immobilized enzyme or microbial cell;... – Enzyme or microbial cell is immobilized on or in an...

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435177, 435183, 435195, 530402, 530811, C12N 1114, C12N 1102, C12N 900, C12N 914

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active

047146761

ABSTRACT:
A naturally occuring protein is chemically modified to provide the protein with activity of a selected enzyme. The protein does not contain activity of the selected enzyme before modification. Modification is preferably carried out by partially denaturing the protein in the presence of an inhibitor for the selected enzyme to form a partially denatured protein-enzyme inhibitor complex, adsorbing the complex to a solid support and crosslinking the adsorbed complex to form an immobilized modified protein having activity of the selected enzyme. Alternatively, the protein may be adsorbed onto the support, partially denatured and crosslinked in the presence of the inhibitor or the protein may be partially denatured, adsorbed on the support and crosslinked in the presence the inhibitor. In another embodiment, the protein has at least three disulphide groups. This protein is contacted with a disulphide bridge reducting agent in the presence of the inhibitor to produce a partially denatured protein-inhibitor complex, the complex is adsorbed onto a support and the adsorbed complex is contacted with a sulfhydryl oxidizing agent to form disulphide bonds in the protein to produce an immobilized modified protein having enzyme activity.

REFERENCES:
patent: 4266030 (1981-05-01), Tschang et al.
patent: 4609625 (1986-09-01), Keyes et al.
Yamauchi, et al., Reversible conversion of Lysine Monoxygenase to an Oxidase, J. of Biol. Chem., vol. 248, 1973, pp. 3750 to 3752.
Mahler et al., Biological Chemistry, 1956, Harper and Row, N.Y. pp. 287-295.
Beaven et al., International Journal of Peptide Research, vol. 5, pp. 215-218, 1973.
Battelle Report, Verification of Semisynthetic Activity, Owens-Illinois, Jul. 14, 1981.

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