Partial purification and characterization of alpha-amylases from Abrus precatorius, Burnatia enneandra and Cadaba farinosa

2014

Article

Klang M.J., Talamond Pascale, Djidimbele N., Tavea F., Ndjouenkeu R.

Journal of Enzyme Research, 2014, 5 (1), p. 66-71 [art. BIA0002353] ISSN 0976-7657

Leaves of Abrus precatorius, tubers of Burnatia enneandra and stems of Cadaba farinosa are used in savannah regions of Cameroon in traditional food processing, particularly in sweetening and liquefaction of gruels. α-amylase was extracted and partially purified from these plants using conventional methods of protein purification including ammonium sulfate fractionation and two steps of gel filtration. Purification achieved 58, 61 and 46 fold respectively for A. precatorius, B. enneandra and C. farinosa. The purified enzymes were then characterized in terms of molecular weight, optimum pH and stability, optimum temperature and stability, Km, Vmax and metals ions effects. The optimum pH of enzymes varied from 6.0 for amylases from B. enneandra and C. farinosa, to 7.0 for amylase from A. precatorius; while the optimum temperature was 60°C for amylases from A. precatorius and B. enneandra, and 65°c for amylase from C. farinosa. The three enzymes displayed, respectively for A. precatorius, B enneandra and C farinosa, a molecular weight of 60, 65 and 48.5 kDa, Km for hydrolyzing soluble starch of 3.25, 1.81 and 3.18 mg/ml, and strong individual activation by Ca2+, Co2+ and Fe3+. Li2+ appeared as a common activator for all the amylases, while Ag+, Hg2 +, Zn2+ and Cu2+ act as common inhibitors.

Sciences fondamentales / Techniques d'analyse et de recherche [020] ; Nutrition, alimentation [054] ; Biotechnologies [084]

Fonds IRD [F B010065248]

fdi:010065248