Partitioning Studies of Alpha-amylase in Aqueous Two Phase System Composing of Polyethylene Glycol and Tri-Sodium Citrate

Subject Areas : Microbiology

مریم  Karimi ¹ (M. Sc. Graduated in Chemical Engineering, Department of Chemical Engineering, Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran.)
N.  Hadidi ² (Assistant Professor of the Department of Clinical Research and Electronic Microscope, Pasteur Institute of Iran, Tehran, Iran.)
Gh.  Pazuki ³ (Associate Professor of the Department of Chemical Engineering, Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran.)
Sh.  Abdolrahimi ⁴ (Ph. D. Graduated in Chemical Engineering, Department of Chemical Engineering, Amirkabir University of Technology (Tehran Polytechnic), Tehran, Iran.)

Received: 2019-10-04 Accepted : 2019-10-04 Published : 2019-09-23

Keywords: &amp, alpha, -Amylase, Aqueous Two Phase System, Poly Ethylene Glycol, Partitioning, Tri Sodium Citrate,

Abstract :

Introduction: One of the scenario for separation of biotechnology products is the application of aqueous two-phase systems (ATPS) rather than traditional extraction by using solvent. The ATPSs are applicable method for separation and purification of biomolecules mixture. The ATPS can be applied for protein purification of biomolecules from suspension broth. The goal of the present research is the study of partitioning of -amylase in polymer-salt aqueous two-phase system composed of polyethylene glycol with molecular weight of 1000 and tri-sodium citrate salt as a biocompatible salt. Materials and Methods: The aim of this research is to study partitioning of α-amylase enzyme in an aqueous two phase system. The system consists of polyethylene glycol (PEG) with molecular weight of 1000 and tri-sodium citrate that is a biocompatible salt. Results: The results for partitioning of enzyme in ATPS has been modeled and statistically analyzed; and indicated that the partition coefficient of -amylase in the system has the maximum amount of 2.681. Conclusion: In this research it has been proved that ATPS components concentration has an effect on the enzyme partition coefficient. Using experimental results, -amylase has a great tendency to be placed in the PEG rich phase.

References:

خیاطی، غ. و علیزاده، ص. (1394). استخراج آنزیم لیپاز تولیدی توسط قارچ Aspergillus niger از براث تخمیری با استفاده از سیستم دوفازی آبی. نشریه علوم مهندسی و جداسازی ، دوره 7، شماره 1، صفحات 53-45.

Cacace, D. N.  & Keating, C. D. (2013). Biocatalyzed mineralization in an aqueous two-phase system: effect of background polymers and enzyme partitioning. Journal of Materials Chemistry B, 1(13), 1767-1870.

Dehnavi, S. M., Pazuki, G. & Vossoughi, M. (2015). PEGylated silica-enzyme nanoconjugates: a new frontier in large scale separation of α-amylase. Scientific Reports, 5, 1-9.

Gomes, G. A., Azevedo, A. M., Aires-Barros, M. R. & Prazeres, D. M. F. (2009). Purification of plasmid DNA with aqueous two phase systems of PEG 600 and sodium citrate/ammonium sulfate. Separation and Purification Technology, 65 (1), 22–30.

Haghtalab, A., Mokhtarani, B. & Maurer, G. (2003). Experimental Results and Thermodynamic Modeling of the Partitioning of Lysozyme, Bovine Serum Albumin, and α-Amylase in Aqueous Two-Phase Systems of PEG and (K₂HPO₄ or Na₂SO₄). Journal of Chemical & Engineering Data, 48 (5), 1170–1177.

 Han, J., Yu, C. & Wang, Y. (2010). Liquid–liquid equilibria of ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate and sodium citrate/tartrate/acetate aqueous two-phase systems at 298.15 K: experiment and correlation. Fluid Phase Equilibria, 295 (1) , 98-103.

Kammoun, R., Naili, B. & Bejar, S. (2008). Application of a statistical design to the optimization of   parameters and culture medium for alpha-amylase production by Aspergillus oryzae CBS 819.72 grown on gruel (wheat grinding by-product). Bioresource Technology, 99 (13), 5602–5609.

Karkas, T. & Önal, S. (2012). Characteristics of invertase partitioned in poly (ethylene glycol)/magnesium sulfate aqueous two-phase system. Biochemical Engineering Journal, 60 (16), 142– 151.

Kavakçıoğlu, B.  & Tarhan, L. (2013). Initial purification of catalase from Phanerochaete chrysosporium by partitioning in poly (ethylene glycol)/salt aqueous two phase systems. Separation and Purification Technology, 105 (22), 8-14.

Khayati, G., Anvari, M. & Shahidi, N. (2015). Partitioning of b-galactosidase in aqueous two-phase systems containing polyethyleneglycol and phosphate salts,Fluid Phase Equilibria, 385 (15), 147-152.

Merchuk, J. C., Andrews B. A. & Asenjo, J. A. (1998). Aqueous two-phase systems for protein separation. Studies on phase inversion. Journal of Chromatography B, 711(1-2), 285–293.

Ozturk, B. (2001). Immobilization of Lipase from Candida rugosa on Hydrophobic and Hydrophilic Supports. Thesis of Department: Biotechnology and Bioengineering. İzmir Institute of Technology İzmir, Turkey.

Sadeghi, R.&  Jahani, F. (2012). Salting-In and Salting-Out of Water-Soluble Polymers in Aqueous Salt Solutions.Journal of Chromatography B, 116 (17), 5234-5241.

Shahriari, S., Neves, C. M., Freire G. M. & Coutinho, J. A. P. (2012). Role of the Hofmeister Series in the Formation of Ionic-Liquid-Based Aqueous Biphasic Systems. The Journal of Physical Chemistry B, 116 (24) , 7252–7258.

Shahriari, S., Vossoughi, M., Taghikhani, V., Safekordi, A. A. & Alemzadeh, I. (2010).  Experimental Study and Mathematical Modeling of Partitioning of β-Amylase and Amyloglucosidase in PEG−Salt Aqueous Two-Phase Systems. Journal of Chemical & Engineering Data, 55 (11), 4968–4975.

Shahriari, S., Vossoughi, M., Taghikhani, V., Safekordi, A. A., Alemzadeh, I. & Pazuki, G. (2012). Measurement of partition coefficients of β-amylase and amyloglucosidase enzymes in aqueous two-phase systems containing poly (ethylene glycol) and Na₂SO₄/KH₂PO₄ at different temperatures. Fluid Phase Equilibria, 292 (30), 80–86.

Subramaniyam, R. & Vimala, R. (2012). Solid state and submerged fermentation for the production of bioactive substances: a comparative study. International Journal of Natural Sciences,3 (3), 480-486.

Tianwei, T., Qing, H. & Qiang, L.  (2002) Purification of glycyrrhizin from Glycyrrhiza uralensis Fisch with ethanol/phosphate aqueous two phase system. Biotechnology Letters, 24(3), 1417–1420.

Xiea, H. G., Wang, Y. J. & Mi Suna, M. (2006). Modeling of the partitioning of membrane protein and phase equilibria for Triton X-100–salt aqueous two-phase systems using a modified generalized multicomponent osmotic virial equation.Process Biochemistry, 41 (3), 689-696.