title: Extraction and purification of beta-amylase from stems of Abrus precatorius by three phase partitioning.
abstract: The stems of Abrus precatorius were used to extract a beta-amylase enriched fraction. A three phase partitioning method and a Doehlert design with 3 variables (ratio of crude extract/t-butanol, the ammonium sulphate saturation and pH) were used. The data was fitted in a second-order polynomial model and the parameters were optimized to enrich beta-amylase. Experimental responses for the modulation were recovery of activity and the purification factor. The optimal conditions were: a ratio of crude extract/t-butanol of 0.87 (v/v), saturation in ammonium sulphate of 49.46% (w/v) and a pH of 5.2. An activity recovery of 156.2% and a purification factor of 10.17 were found. The enriched enzyme was identified as a beta-amylase and its molecular weight was 60.1kDa. Km and Vmax values were 79.37mg/ml and 5.13U/ml, respectively and the highest activity was registered at a temperature of 70°C and a pH between 6 and 6.5. A significant stabilization of the beta-amylase was observed up to 65°C.
authors: Department of Process Engineering, National School of Agro-Industrial Sciences (ENSAI), University of Ngaoundere, P.O. Box 455, Adamaoua, Cameroon.
authors: Sorel Tchewonpi
authors: ST
authors: Sagu
authors: Department of Process Engineering, National School of Agro-Industrial Sciences (ENSAI), University of Ngaoundere, P.O. Box 455, Adamaoua, Cameroon.
authors: Emmanuel Jong
authors: EJ
authors: Nso
authors: Instrumental Analysis in Nutritional Science, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany.
authors: Thomas
authors: T
authors: Homann
authors: Department of Process Engineering, National School of Agro-Industrial Sciences (ENSAI), University of Ngaoundere, P.O. Box 455, Adamaoua, Cameroon.
authors: César
authors: C
authors: Kapseu
authors: Instrumental Analysis in Nutritional Science, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany. Electronic address: rawel@uni-potsdam.de.
authors: Harshadrai M
authors: HM
authors: Rawel
mesh_terms: D029912:Abrus / Q000737:chemistry*; D000645:Ammonium Sulfate; D005591:Chemical Fractionation / Q000379:methods*; D013696:Temperature; D001614:beta-Amylase / Q000737:chemistry*
chemical_list: D001614:beta-Amylase; D000645:Ammonium Sulfate
keywords: Abrus precatorius; Beta-amylase; Doehlert design; Purification; Three phase partitioning