Extraction of Erythrocyte Enzymes for the Preparation of Polyhemoglobin-catalase-superoxide Dismutase

Figures & data

Scheme 1 Relationship between xanthine oxidase (XOD), superoxide dismutase (SOD), catalase (CAT), superoxide radical (O₂⁻), and hydrogen peroxide (H₂O₂).

Table 1. Catalase and SOD activity after cross-linking with hemoglobin in PolyHb, PolySFHb and PolyHb-SOD-CAT

Sample	Hb (g): SOD (U): CAT (U)
PolyHb	1:860:240
SFHb	1:12000:33800
PolySFHb	1:10800:23000
PolyHb-SOD-CAT	1:15060:330000

Figure 1.  Peroxidase-catalyzed measurement of hydrogen peroxide 3 min after the addition of 200uM of hydrogen peroxide in polySFHb solution (Catalase: 23.0 U/mg Hb).

Figure 2.  Catalase activity retained following polymerization.

Figure 3.  Hydrogen peroxide measured, 3 min after the addition of 200uM of hydrogen peroxide in polySFHb-CAT solution (1 uM heme) with different catalase concentration.

Figure 4.  Conversion NBT to NBT-diformazan, by xanthine/xanthine oxidase–generated superoxides radical(O₂⁻), in the presence of PolyHb or PolySFHb or PolySFHb-SOD-CAT (1 µM heme).

Figure 5.  Absorbance spectra of hemoglobin (10µM) in PolyHb following H₂O₂ addition of 10, 100, 500, and 1000 µM.

Figure 6.  Absorbance spectra of hemoglobin (10µM) in PolySFHb following H₂O₂ addition of 10, 100, 500, and 1000 µM.

Figure 7.  Absorbance spectra of hemoglobin (10µM) in PolyHb-SOD-CAT following H₂O₂ addition of 10, 100, 500, and 1000 µM.

Figure 8.  Absorbance spectra of reaction mixtures containing 10 uM of hemoglobin and 2.0 mM of glucose, and 0.2 U/ml glucose oxidase for continuous generation of hydrogen peroxide. Top: PolyHb. Lower left: PolySFHb, Lower right: PolySFHb-SOD-CAT.

Figure 9.  Absorbance spectra of reaction mixtures containing 10 uM of hemoglobin and 100 uM of xanthine, and 10 mU/ml xanthine oxidase for continuous generation of superoxide. Top: PolyHb. Lower left: PolySFHb. Lower right: PolySFHb-SOD-CAT.

Figure 10.  Ferrylhemoglobin formation in the mixture containing 10 µM (heme) of PolyHb, PolySFHb, and PolySFHb-SOD-CAT in the presence of hydrogen peroxide (Left: 100 µM and right: 500 µM).

Figure 11.  Ferrylhemoglobin formation in the mixture containing 10 µM (heme) of PolyHb, PolySFHb or PolySFHb-SOD-CAT in the presence of (left) glucose (2 mM) /glucose oxidase (0.2 U) and (right) Xanthine (100 µM) /xanthine oxidase (10 mU).