Optimization of Borassus flabellifer Amylase Extraction Procedure Using Box-Behnken Design and Development of Simple Affinity Chromatographic Technique for Purification of Amylases.
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Date
2012-07
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Abstract
This work is aimed to study the optimal conditions of extraction and development of novel
purification process for α-Amylase from pericarp of Borassus flabellifer fruit. Optimization
was done by Response Surface Methodology (RSM) using Box-Behnken design. The
variables chosen for this purpose are pH (X1), temperature (X2, °C) and CaCl2
concentration (X3, ppm). A total of 9 runs were conducted, varying each factor while
others were kept constant. The contribution of each factor was established giving raise to
significant t-values and p-values. This revealed that the three variables are essential for
activity of the enzyme. Based on the obtained optimal conditions from RSM studies,
enzyme formation was successfully achieved with an activity of 28.8 U and concentration
of 1.4 mg/ml for the 90% ammonium sulfate precipitated dialyzed enzyme sample. A
novel affinity chromatographic process was developed for purification process and by this process the enzyme concentration was found to be 0.032mg/ml and activity was
determined as 4.76 U.
Description
Keywords
Amylase, Borassus, partial purification, RSM, Box-Behnken, affinity chromatography
Citation
Bakku Ranjith Kumar, Terli Ramana, Somalanka Subba Rao, Palukurty Mary Anupama. Optimization of Borassus flabellifer Amylase Extraction Procedure Using Box-Behnken Design and Development of Simple Affinity Chromatographic Technique for Purification of Amylases. British Biotechnology Journal. 2012 July; 2(3): 146-156.