The very first example of benign intercalation of met-hemoglobin (Hb) in layered double hydroxide, LDH (anionic layered material, Mg₃Al-NO₃), in two different buffers (10 mM Tris-HCl pH 7.2 and 10 mM dibasic K₂HPO4 buffer pH 7.2) is described here. The binding constant of Hb for LDH in phosphate buffer is 100 times greater than the binding constant in Tris-HCl (1.7 x 10³ /M). Circular Dichroism studies indicated significant retention of the secondary structure of bound protein (>90%). However, in contrast to the structure retention, the peroxidase-like activity of intercalated Hb is lower than in solution. The V_max for the bound protein, for example, is 10 times lower than the corresponding V_max of the free Hb (1.6 x 10^-3 mM/s in phosphate buffer). Hb/LDH complex in phosphate buffer showed greater activity as compared to the complex in Tris buffer (> 2 folds in presence of 5 mM substrate, 1 mM H₂O₂). The differential scanning calorimetric thermogram of Hb shows increased resistance to denaturation of the bound protein. The denaturation enthalpy, for example, for the bound protein (in 10 mM K₂HPO₄ pH 7.2) is approx. 3 folds greater than the corresponding enthalpy of the free Hb (152 ± 2 kcal/mol). Furthermore, the increase of enthalpy is compensated by the change in entropy. The current studies clearly indicates the formation and characterization of novel biomatrices and such approaches might form the basis for engineering novel biomaterials for applications as biosensors and biocatalysis, with improved properties.