Bienzymatic supramolecular complex of catalase modified with cyclodextrin-branched carboxymethylcellulose and superoxide dismutase: stability and anti-inflammatory properties.

A bienzymatic supramolecular assembly of CAT and SOD is reported. CAT was chemically glycosilated with CD branched CMC and then associated with SOD modified with 1-adamantane carboxylic acid. SOD was remarkably resistant to inactivation by H(2)O(2) and its anti-inflammatory activity was 4.5-fold increased after supramolecular association with the modified CAT form. [structure: see text]

Improved pharmacological properties for superoxide dismutase modified with beta-cyclodextrin-carboxymethylcellulose polymer.

Superoxide dismutase was glycosidated with cyclodextrin-branched carboxymethylcellulose. The modified enzyme contained 1.4 mol polymer per mol protein and retained 87% of the initial activity. The anti-inflammatory activity of superoxide dismutase was 2.2-times increased after conjugation and its plasma half-life time was prolonged from 4.8 min to 7.2 h.

Pharmacokinetics and stability properties of catalase modified with water-soluble polysaccharides.

Bovine liver catalase (EC 1.11.1.6) was chemically modified with mannan, carboxymethylcellulose, and carboxymethylchitin. The enzyme retained about 48-97% of the initial specific activity after glycosidation with the polysaccharides. The prepared neoglycoenzyme was 1.9-5.7 fold more stable against the thermal inactivation processes at 55 degrees C, in comparison with the native counterpart. Also, the modified enzyme was more resistant to proteolytic degradation with trypsin. Pharmacokinetics studies revealed higher plasma half-life time for all the enzyme-polymer preparations, but better results were achieved for the enzyme modified with the anionic macromolecules.

Improved pharmacological properties for superoxide dismutase modified with mannan

Mannan from Sacharomyces cerevisiae was activated by oxidation with NaIO4 (sodium m-periodate) and further linked to SOD (superoxide dismutase) via reductive alkylation with NaBH4 (sodium borohydride). The gly-cosidated enzyme contained an average of 1,2 mol of polysaccharide per mol of protein and retained 52 per cent of its initial activity. The modified enzyme was 560-fold more resistant to inactivation with H2O2 and acquired a lectin-recognition capacity in respect of concanavalin A. The anti-inflammatory activity of SOD was increased 2-fold and its plasma half-life time was prolonged from 4.8 min to 1.7 h after glycosylation with the polymer(AU)

Improved pharmacological properties for superoxide dismutase modified with mannan.

Mannan from Sacharomyces cerevisiae was activated by oxidation with NaIO(4) (sodium m-periodate) and further linked to SOD (superoxide dismutase) via reductive alkylation with NaBH(4) (sodium borohydride). The glycosidated enzyme contained an average of 1.2 mol of polysaccharide per mol of protein and retained 52% of its initial activity. The modified enzyme was 560-fold more resistant to inactivation with H(2)O(2) and acquired a lectin-recognition capacity in respect of concanavalin A. The anti-inflammatory activity of SOD was increased 2-fold and its plasma half-life time was prolonged from 4.8 min to 1.7 h after glycosylation with the polymer.

Transglutaminase-catalyzed site-specific glycosidation of catalase with aminated dextran.

An enzymatic approach, based on a transglutaminase-catalyzed coupling reaction, was investigated to modify bovine liver catalase with an end-group aminated dextran derivative. We demonstrated that catalase activity increased after enzymatic glycosidation and that the conjugate was 3.8-fold more stable to thermal inactivation at 55 degrees C and 2-fold more resistant to proteolytic degradation by trypsin. Moreover, the transglutaminase-mediated modification also improved the pharmacokinetics behavior of catalase, increasing 2.5-fold its plasma half-life time and reducing 3-fold the total clearance after its i.v. administration in rats.

Glycosidation of cu,zn-superoxide dismutase with end-group aminated dextran: pharmacological and pharmacokinetics properties.

Bovine Cu,Zn-SOD was chemically modified with an end-group aminated dextran derivative using a water-soluble carbodiimide as coupling agent. The enzyme retained 81% of the initial catalytic activity after the attachment of about 4.4 mol of polymer per protein subunit. The anti-inflammatory activity of the SOD was two times increased after conjugation with dextran. The modified enzyme was remarkably more resistant to inactivation by H(2)O(2) and its plasma half-life time was prolonged from 4 min to 3.2 h.

Increased conformational and thermal stability properties for phenylalanine dehydrogenase by chemical glycosidation with end-group activated dextran.

A mono-aminated dextran derivative was attached to Bacillus badius phenylalanine dehydrogenase via a carbodiimide-catalyzed reaction. The optimum temperature for the conjugate was 10 degrees C higher than for native enzyme, and its thermostability was improved by 8 degrees C. The activation free energy of thermal inactivation at 45 degrees C was increased by 16.8 kJ/mol. The improved conformational stability of the modified enzyme was confirmed by fluorescence spectroscopy.

Improved anti-inflammatory and pharmacokinetic properties for superoxide dismutase by chemical glycosidation with carboxymethylchitin.

O-carboxymethylchitin (molecular weight = 1.07 x 10(5), degree of carboxymethylation = 80%, degree of N-acetylation = 91%) was chemically attached to superoxide dismutase by the formation of amide linkages through a carbodiimide catalyzed reaction. The glycosidated enzyme contained about 1.8 mole of polysaccharide per mole of protein and retained 57% of the initial catalytic activity. The anti-inflammatory activity of the enzyme was 2.4 times increased after conjugation with the polysaccharide. The modified superoxide dismutase preparation was remarkably more resistant to inactivation with H(2)O(2) and its plasma half-life time was prolonged from 4.8 min to 69 h.