Optimization of Cutinase Purification using a Hydrophobic Interaction Membrane Chromatographic Process by Response Surface Methodology

Abdul Wahab Mohammad, Suhaila Johar, Jamaliah Md Jahim, Osman Hassan

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1 Citation (Scopus)

Abstract

Several methods have been demonstrated in the recovery and purification of cutinase; however, hydrophobic interaction membrane chromatography (HIMC) has yet to be implemented for this purpose. In this study, two factors, that is, buffer pH and ammonium sulphate concentration for HIMC operation, were optimized using response surface methodology (RSM), aiming to obtain the highest cutinase purification factor and recovery possible. The experimental design used was a central composite design (CCD) comprised of 13 experimental runs. The analysis of variance (ANOVA) indicated that a quadratic model best represented the system. Based on the R2 values, it can be concluded that around 90% of the variability in the system can be explained by the fitted models. The optimized HIMC conditions were buffer pH 6.0 and 1.3 M ammonium sulphate, which was expected to provide 27-fold cutinase purification factor and 85% recovery. However, the actual values deviated from the predicted values with 13% and 18% error, respectively. The statistical analysis showed that only the ammonium sulphate concentration had a great influence on cutinase purification factor. Meanwhile, for cutinase recovery, both parameters were important in determining the optimal condition for cutinase adsorption and desorption to the chromatographic media.

Original languageEnglish
Pages (from-to)1923-1929
Number of pages7
JournalSeparation Science and Technology (Philadelphia)
Volume48
Issue number13
DOIs
Publication statusPublished - Sep 2013

Fingerprint

Purification
Chromatography
Membranes
Recovery
Ammonium Sulfate
Buffers
Analysis of variance (ANOVA)
Design of experiments
Desorption
Statistical methods
Adsorption
cutinase
Composite materials
Sulfates

Keywords

  • cutinase
  • hydrophobic interaction
  • membrane chromatography
  • purification
  • response surface methodology

ASJC Scopus subject areas

  • Chemistry(all)
  • Process Chemistry and Technology
  • Chemical Engineering(all)
  • Filtration and Separation

Cite this

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abstract = "Several methods have been demonstrated in the recovery and purification of cutinase; however, hydrophobic interaction membrane chromatography (HIMC) has yet to be implemented for this purpose. In this study, two factors, that is, buffer pH and ammonium sulphate concentration for HIMC operation, were optimized using response surface methodology (RSM), aiming to obtain the highest cutinase purification factor and recovery possible. The experimental design used was a central composite design (CCD) comprised of 13 experimental runs. The analysis of variance (ANOVA) indicated that a quadratic model best represented the system. Based on the R2 values, it can be concluded that around 90{\%} of the variability in the system can be explained by the fitted models. The optimized HIMC conditions were buffer pH 6.0 and 1.3 M ammonium sulphate, which was expected to provide 27-fold cutinase purification factor and 85{\%} recovery. However, the actual values deviated from the predicted values with 13{\%} and 18{\%} error, respectively. The statistical analysis showed that only the ammonium sulphate concentration had a great influence on cutinase purification factor. Meanwhile, for cutinase recovery, both parameters were important in determining the optimal condition for cutinase adsorption and desorption to the chromatographic media.",
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