Superoxide dismutase, its immobilized form, and their production and use

Claim 2

Superoxide dismutase having the following properties: (a) Activity: to dismutate a superoxide ion into a hydrogen peroxide molecule and an oxygen molecule; (b) Substrate specificity: to act on superoxide ion; (c) Optimum pH range: about 7 to 8; (e) Stable pH range: about 6 to 11; (e) Optimum functional temperature range: about 20° to 45° C.; (f) Thermostability: stable at 37° C. for 60 minutes; Inactivated about 50% at 50°-60° C. in 60 minutes; Inactivated about 100% at 80° C. in 5 minutes; (g) Molecular weight: about 4.8×10 4 (by gel filtration method); (h) Elemental analysis: C. 48.90±2.0%, H. 7.05±2.0%, N. 14.80±2.0%, (i) Absorptions in the visible region of the spectrum; 470-475 nm (max.), 375-380 nm (min.), about 610 nm (shoulder); (j) Molar extinction coefficient: about 1.08×10 5 M -1 .cm -1 at 280 nm; (k) Amino acid analysis: ______________________________________ Amino acid Number of residues ______________________________________ Glycine 23.5 Leucine 28.3 ______________________________________ wherein the values denote the number of amino acid residues per molecule of the enzyme.

Claim 3

An antiinflammatory agent which contains an effective amount of superoxide dismutase having the following properties: (a) Activity: to dismutate a superoxide ion into a hydrogen peroxide molecule and an oxygen molecule; (b) Substrate specificity: to act on superoxide ion; (c) Optimum pH range: about 7 to 8; (d) Stable pH range: about 6 to 11; (e) Optimum functional temperature range: about 20° to 45° C.; (f) Thermostability: Stable at 37° C. for 60 mintues; Inactivated about 50% at 50°-60° C. in 60 minutes; Inactivated about 100% at 80° C. in 5 minutes; (g) Molecular weight: about 4.8×10 4 (by gel filtration method); (h) Elemental analysis: C. 48.90±2.0%, H. 7.05±2.0%, N. 14.8±2.0%, (i) Absorptions in the visible region of the spectrum: 470-475 nm (max.), 375-380 nm (min.), about 610 nm (shoulder); (j) Molar extinction coefficient: approx. 1.08×10 5 M -1 . cm -1 at 280 nm; (k) Amino acid analysis: ______________________________________ Amino acid Number of residues ______________________________________ Glycine 23.5 Leucine 28.3 ______________________________________ wherein the values denote the number of amino acid residues per molecule of the enzyme, as an active ingredient in association with a pharmaceutically acceptable carrier or excipient therefor.

Claim 4

An antiinflammatory agent which contains an effective amount of superoxide dismutase having the following properties: (a) Activity: to dismutate a superoxide ion into a hydrogen peroxide molecule and an oxygen molecule; (b) Substrate specificity: to act on superoxide ion; (c) Optimum pH range: about 7 to 8; (d) Stable pH range: about 6 to 11; (e) Optimum functional temperature range: about 20° to 45° C.; (f) Thermostability: Stable at 37° C. for 60 mintues; Inactivated about 50% at 50°-60° C. in 60 minutes; Inactivated about 100% at 80° C. in 5 minutes; (g) Molecular weight: about 4.8×10 4 (by gel filtration method); (h) Elemental analysis: C. 48.90±2.0%, H. 7.05±2.0%, N. 14.8±2.0%, (i) Absorptions in the visible region of the spectrum: 470-475 nm (max.), 375-380 nm (min.), about 610 nm (shoulder); (j) Molar extinction coefficient: approx. 1.08×10 5 M -1 . cm -1 at 280 nm; (k) Amino acid analysis: ______________________________________ Amino acid Number of residues ______________________________________ Glycine 23.5 Leucine 28.3 ______________________________________ wherein the values denote the number of amino acid residues per molecule of the enzyme, as an active ingredient in association with a pharmaceutically acceptable carrier or excipient therefor.

Claim 5

An immoblized superoxide dismutase which is produced by reacting superoxide dismutase having the following properties: (a) Activity: to dismutate a superoxide ion into a hydrogen perioxide molecule and an oxygen molecule; (b) Substrate specificity: to act on superioxide ion; (c) Optimum pH range: about 7 to 8; (d) Stable pH range: about 6 to 11; (e) Optimum functional temperature range: about 20° to 45° C.;

Claim 6

An immoblized superoxide dismutase which is produced by reacting superoxide dismutase having the following properties: (a) Activity: to dismutate a superoxide ion into a hydrogen perioxide molecule and an oxygen molecule; (b) Substrate specificity: to act on superioxide ion; (c) Optimum pH range: about 7 to 8; (d) Stable pH range: about 6 to 11; (e) Optimum functional temperature range: about 20° to 45° C.;

Claim 7

(f) Thermostability: stable at 37° C. for 60 minutes; Inactivated about 50% at 50°-60° C. in 60 minutes; Inactivated about 100° at 80° C. in 5 minutes; (g) Molecular weight: about 4.8×10 4 (by gel filtration method); (h) Elemental analysis: C. 48.90±2.08%, H. 7.05±2.08%, N. 14.80±2.08%, (i) Absorptions in the visible region of the spectrum: 470-475 nm (max.), 375-390 nm (min.), about 610 nm (shoulder); (j) Molar extinction coefficient: about 1.08×10 5 M -1 . cm -1 at 280 nm; (k) Amino acid analysis: ______________________________________ Amino acid Number of residues ______________________________________ Glycine 23.5 Leucine 28.3 ______________________________________ the values denote the number of amino acid residues per molecule of the enzyme, with a water-soluble carrier.

Claim 8

(f) Thermostability: stable at 37° C. for 60 minutes; Inactivated about 50% at 50°-60° C. in 60 minutes; Inactivated about 100° at 80° C. in 5 minutes; (g) Molecular weight: about 4.8×10 4 (by gel filtration method); (h) Elemental analysis: C. 48.90±2.08%, H. 7.05±2.08%, N. 14.80±2.08%, (i) Absorptions in the visible region of the spectrum: 470-475 nm (max.), 375-390 nm (min.), about 610 nm (shoulder); (j) Molar extinction coefficient: about 1.08×10 5 M -1 . cm -1 at 280 nm; (k) Amino acid analysis: ______________________________________ Amino acid Number of residues ______________________________________ Glycine 23.5 Leucine 28.3 ______________________________________ the values denote the number of amino acid residues per molecule of the enzyme, with a water-soluble carrier.

Claim 9

An immobilized superoxide dismutase according to claim 3, wherein the water-soluble carrier is water-soluble polysaccharide.

Claim 10

An immobilized superoxide dismutase according to claim 4, wherein the water-soluble carrier is dextran or Ficoll.

Claim 11

An antiinflammatory agent which contains an effective amount of an immbolized superoxide dismutase which is produced by reacting superoxide dismutase having the following properties: (a) Activity: to dismutate a superoxide ion into a hydrogen peroxide molecule and an oxygen molecule; (b) Substrate specificity: to act on superoxide ion; (c) Optimum pH range: about 7 to 8; (d) Stable pH range: about 6 to 11; (e) Optimum functional temperature range: about 20° to 45° C.; (f) Thermostability: Stable at 37° C. for 60 minutes; Inactivated about 50% at 50°-60° C. in 60 minutes; Inactivated about 100% at 80° C. in 5 minutes; (g) Molecular weight about 4.8×10 4 (by gel filtration method); (h) Elemental analysis: C. 48.90±2.0%, H. 7.05±2.0%, N. 14.80±2.0%, (i) Absorptions in the visible region of the spectrum: 470-475 nm (max.), 375-380 nm (min.), about 610 nm (shoulder); (j) Molar extinction coefficient: approx. 1.08×10 5 M -1 . cm -1 at 280 nm; (k) Amino acid analysis: ______________________________________ Amino acid Number of residues ______________________________________ Glycine 23.5 Leucine 28.3 ______________________________________ wherein the values denote the number of amino acid residues per molecule of the enzyme, with a water-soluble carrier, as an active ingredient in association with a pharmaceutically acceptable carrier or excipient therefor.

Claim 12

An antiinflammatory agent which contains an effective amount of an immbolized superoxide dismutase which is produced by reacting superoxide dismutase having the following properties: (a) Activity: to dismutate a superoxide ion into a hydrogen peroxide molecule and an oxygen molecule; (b) Substrate specificity: to act on superoxide ion; (c) Optimum pH range: about 7 to 8; (d) Stable pH range: about 6 to 11; (e) Optimum functional temperature range: about 20° to 45° C.; (f) Thermostability: Stable at 37° C. for 60 minutes; Inactivated about 50% at 50°-60° C. in 60 minutes; Inactivated about 100% at 80° C. in 5 minutes; (g) Molecular weight about 4.8×10 4 (by gel filtration method); (h) Elemental analysis: C. 48.90±2.0%, H. 7.05±2.0%, N. 14.80±2.0%, (i) Absorptions in the visible region of the spectrum: 470-475 nm (max.), 375-380 nm (min.), about 610 nm (shoulder); (j) Molar extinction coefficient: approx. 1.08×10 5 M -1 . cm -1 at 280 nm; (k) Amino acid analysis: ______________________________________ Amino acid Number of residues ______________________________________ Glycine 23.5 Leucine 28.3 ______________________________________ wherein the values denote the number of amino acid residues per molecule of the enzyme, with a water-soluble carrier, as an active ingredient in association with a pharmaceutically acceptable carrier or excipient therefor.

Claim 13

An antiinflammatory agent according to claim 6, wherein the water-soluble carrier is a water-soluble polysaccharide.

Claim 14

An antiinflammatory agent according to claim 7, wherein the water-soluble carrier is dextran or Ficoll.