1.4.3.3: D-amino-acid oxidase

391839

carrier-free enzyme is entrapped in semipermeable microcapsules produced from the polycation poly(methylene-co-guanidine) in combination with CaCl2 and the polyanions alginate and cellulose sulfate, the effectiveness of the entrapped oxidase for O2-dependent conversion of D-methionine at 25°C is 75-95% of the free enzyme preparation

685713

cetylpyridinium bromide stabilizes

391852

changes in ionic strength, I: 16-82 mM, have only moderate effect on enzyme stability, stability maximum is 50 mM

391833

chymotrypsin, 10% w/w, no effect

Sus scrofa

391836

covalent modification of DAO using maleimide-activated PEG5000 yields a stable bionconjugate in which three exposed cysteine side chains are derivatized, the PEGylated enzyme shows approximately 3.3fold slowed dissociation of the FAD cofactor at 50°C, and this causes a 2fold thermostabilization of the enzyme activity

685045

D-amino acid oxidase fused to an elastin-like polypeptide exhibits enzymatic activity that is about 1.6times that of the free enzyme and shows higher stability

742950

DAAO immobilized on a macro-porous carrier is higher in water-insoluble organic solvents than in water-soluble ones

674970

enzyme immobilized on Amberzyme oxirane resin can be recycled more than 14 times without significant loss of activity

671518

glycerol, 10%, stabilizes

391833, 391852

glycerol, 50% stabilizes

Candida tropicalis

391834

good stability to urea denaturation

673646

His-tagged DAAO loses its flavin cofactor upon dilution or prolonged dialysis against FAD-free solvents leading to the catalytic inactivation

Q99042

685355

homodimer is stable even in the apoprotein form

677016

immobilisation on PEI 25 kDa Sepabeads and treatment with 0.5% glutaraldehyde solution at pH 7-8 and 25°C for 16 h leads to 160fold increased enzyme stability

Q99042

686277

incubation of the mutant enzyme DELTASer308-Lys321 with 10% trypsin in presence of exogenous FAD, 0.2 mM, shows that the rate of activity loss is lower than that determined in absence of exogenous FAD

P80324

655558

m-toluic acid stabilizes

391839

multisubunit immobilization on highly activated glyoxyl agarose improves stability of the enzyme 1500fold, at a protein concentration of 0.0067 mg/ml.

655115

multisubunit immobilization on highly activated glyoxyl agarose marginally improves stability of the enzyme, 15-20fold, at a protein concentration of 0.0067 mg/ml. Dissociation of FAD is not prevented by immobilization

655115

oriented immobilization of a chimeric oxidase maintains 80% of the original activity in microparticle-bound enzymes. Mildly permeabilized cells in which N-terminally tagged TvDAO in which a 12-amino-acid peptide is fused in frame to the N-terminus (TvDAOstrepN) is physically entrapped are employed for the batchwise conversion of D-Ala in an aerated enzyme reactor. The cells could be reused for at least three rounds of reaction with only small losses of activity. The free enzyme, however, is rapidly inactivated under the same reaction conditions (half-life: 1.2 h)

671491

stabilization of the enzyme by immobilization onto Fe3O4 magnetic nanoparticles, after immobilization the Tm increases from 45°C of the free form to 55°C. In the presence of 20 mM H2O2, the immobilized form retains 93% of its activity after 5 h while the free form is completely inactivated after 3.5 h

P80324

696910

stable homodimer even in the apoprotein form

673646

stable to guanidine

391866

the catalytic efficiency of immobilized DAO (onto streptavidin-coated magnetic beads through the interaction between biotin and streptavidin) toward D-alanine is decreased by 56%

2 entries

the half-life of the enzyme can be up to 15.5 days by treating the cetyltrimethylammonium bromide-permeabilized cells with 1% (w/v) glutaraldehyde

Komagataella pastoris

723900

the midpoint contration of urea required for unfolding is 1.4-1.8 M for wild-type enzyme and 0.8-1.1 M for mutant enzyme W243Y

P80324

673567

the surfactant Pluronic F-68 stabilizes immobilized DAO by protecting the enzyme from the deleterious effect of gas-liquid interfaces