Purification and characterization of chondroitinase ABC from Acinetobacter sp. C26

- Purification and chondroitinase ABC from Acinetobacter sp. C26 was carried out.
- Identification of purified chondroitinase ABC was determined.
- Characterisation of the enzyme was carried out with respect to pH, temperature and presence of metal ions and chemicals.
- The kinetic parameters of the enzyme were studied.

An extracellular chondroitinase ABC (ChSase ABC, EC 4.2.2.4) produced by cultivating Acinetobacter sp. C26, was purified to homogeneity from the supernatant by ammonium sulfate fractionation, Q-Sepharose Fast Flow and Sephadex G-100 chromatography. The 76 kDa enzyme was purified 48.09-fold to homogeneity with specific activity of 348.64 U/mg, Using the chondroitin sulfate A (CS-A) as substrate, the maximal reaction rate (Vmax) and Michaelis-Menten constant (Km) of ChSase ABC were found to be 10.471 μmol/min/ml and 0.105 mg/ml, respectively. The enzyme showed the highest activity at the optimal conditions of pH 6.0 and 42 ∘C, respectively. This enzyme was stable at pH 5-10, 5-9 and 5-7 at 4 °C, 37 °C and 42 °C, respectively. Investigation about thermal stability of ChSase ABC displayed that it was stable at 37 °C. ChSase ABC activity was increased in presence of Na+, K+, Mn2+, 1,10-phenanthrolin and strongly inhibited by Cu2+, Hg2+, Al3+and SDS. These properties suggested that ChSase ABC from Acinetobacter sp. C26 bring promising prospects in medical and industry applications.