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Research Article Open access CC BY 3.0

Optimization of Borassus flabellifer Amylase Extraction Procedure Using Box-Behnken Design and Development of Simple Affinity Chromatographic Technique for Purification of Amylases

Ranjith Kumar Bakku, Ramana Terli, Subba Rao Somalanka, Mary Anupama Palukurty

Biotechnology Journal International · pp. 146–156 · Published 13 Sep 2012

10.9734/BBJ/2012/1200

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.

Amylase Borassus partial purification RSM Box-Behnken affinity chromatography

References (17)

  1. 1 PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT [DOI]
  2. 2 Some New Three Level Designs for the Study of Quantitative Variables [DOI]
  3. 3 Response Surfaces: Designs and Analyses [DOI]
  4. 4 Chemometrics: Experimental design [DOI]
  5. 5 Chemometrics for experimental design [DOI]
  6. 6 Amylases and their applications [DOI]
  7. 7 Use of Box-Behnken design of experiments for the adsorption of verofix red using biopolymer [DOI]
  8. 8 Isolation and characterization of a novel thermostable α-amylase from Korean pine seeds [DOI]
  9. 9 Purification of a bifunctional amylase/protease inhibitor from ragi (Eleusine coracana) by chromatography and its use as an affinity ligand [DOI]
  10. 10 Response surface methodology to evaluation the recovery of amylases by hollow fiber membrane [DOI]
  11. 11 Optimization of medium constituents for Cephalosporin C production using response surface methodology and artificial neural networks
  12. 12 Partial Characterization of α-Amylase from Germinating Little Millets (Panicum sumatrense)
  13. 13 Purification and properties of a starch granule-degrading α-amylase from potato tubers [DOI]
  14. 14 Studies on the extraction and characterization of thermostable α-amylase from pericarp of Borassus indica
  15. 15 Affinity chromatography of mustard β-amylase on starch columns [DOI]
  16. 16 Affinity Binding of Higher Plant β‐Amylases to Starch. A Rapid Purification Method [DOI]
  17. 17 Optimization of malting conditions of Pearl millet (Pennisetum glaucum (L.)R.Br.) using Box-Behnken design

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