Single Step Purification of Bromelain from Ananas Comosus Residues by PEG/Ammonium Sulphate Integrated Aqueous Two-Phase Systems


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Abstract


This work evaluates a single step purification of bromelain using a PEG/Ammonium Sulphate integrated aqueous two-phase systems (ATPS). This new process integrates the ammonium sulphate precipitation of unwanted proteins along with purification of target molecule by ATPS. The results, analysed by response surface methodology, showed that higher values of protein yield and enzyme activity recovery could be achieved by desirability optimization. Thus, it was possible to obtain a biological activity recovery over than 87.3% with a purification factor of 11.8 fold, using a system composed by 10.86% PEG 4,000 and 36.21% ammonium sulphate saturation. The purified bromelain showed an increased stability when supplemented with PEG, without any loss of activity after 7 hours of incubation
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Keywords


Bromelain; Ananas Comosus; Aqueous Two-Phase Systems; Purification; Stability

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References


R. M. Heinicke, Complementary Enzyme Actions in the Clotting of Milk, Science 118 (1953) 753-754.

H. R. Maurer, Bromelain: biochemistry, pharmacology and medical use, Cellular and Molecular Life Sciences (CMLS) 58 (2001) 1234-1245.

E. R. Secor Jr, W. F. Carson Iv, M. M. Cloutier, L. A. Guernsey, C. M. Schramm, C. A. Wu, R. S. Thrall, Bromelain exerts anti-inflammatory effects in an ovalbumin-induced murine model of allergic airway disease, Cellular Immunology 237 (2005) 68-75.

T. Harrach, K. Eckert, H. R. Maurer, I. Machleidt, W. Machleidt, R. Nuck, Isolation and characterization of two forms of an acidic bromelain stem proteinase, Journal of Protein Chemistry 17 (1998) 351-361.

O. S. Chatsworth, Cosmetic composition containing alpha hydroxyacids, salicyclic acid, and enzyme mixture of bromelain and papain, Biotechnology Advances 14 (1996) 562.

E. Silveira, M. E. Souza-Jr, J. C. C. Santana, A. C. Chaves, A. L. F. Porto, E. B. Tambourgi, Expanded bed adsorption of bromelain (E.C. 3.4.22.33) from Ananas comosus crude extract, Brazilian Journal of Chemical Engineering 26 (2009) 149-157.

J. F. Ferreira, J. C. C. Santana, E. Tambourgi, The effect of pH on bromelain partition from Ananas comosus by PEG4000/phosphate ATPS, Brazilian Archives of Biology and Technology 54 (2011) 125-132.

P. Cutler, Protein purification protocols. Vol 244. (2nd ed. Totowa, New Jersey: Humana Press; 2004, pp. 91-99).

A. Narayan, M. Madhusudhan, K. Raghavarao, Extraction and Purification of Ipomoea Peroxidase Employing Three-phase Partitioning, Applied Biochemistry and Biotechnology 151 (2008) 263-272.

Sigma-Chemical-Company, Catalog of Biochemical and chemical reagents (2006).

M. M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry 72 (1976) 248-254.

L. A. Oliveira, A. L. F. Porto, E. B. Tambourgi, Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes, Bioresource Technology 97 (2006) 862-867.

M. P. Deutscher, ed. Guide to protein purification. Vol 182. (San Diego, USA: Academic Press; 1990, pp. 285-300).

P. Soares, D. Coelho, P. Mazzola, E. Silveira, M. D. G. Carneiro-da-Cunha, A. Pessoa, E. Tambourgi, Studies on bromelain precipitation by Ethanol, poly (Ethylene Glycol) and Ammonium Sulphate, Chemical Engineering Transactions 24 (2011) 979-984.

R. V. Devakate, V. V. Patil, S. S. Waje, B. N. Thorat, Purification and drying of bromelain, Separation and Purification Technology 64 (2009) 259-264.

H. Umesh Hebbar, B. Sumana, K. S. M. S. Raghavarao, Use of reverse micellar systems for the extraction and purification of bromelain from pineapple wastes, Bioresource Technology 99 (2008) 4896-4902.

K. Ratanapongleka, Recovery of Biological Products in Aqueous Two Phase Systems, International Journal of Chemical Engineering and Applications 1 (2010) 191-198.

D. P. Harris, A. T. Andrews, G. Wright, D. L. Pyle, J. A. Asenjo, The application of aqueous two-phase systems to the purification of pharmaceutical proteins from transgenic sheep milk, Bioseparation 7 (1998) 31-37.

S. Fatima, R. H. Khan, Effect of Polyethylene Glycols on the Function and Structure of Thiol Proteases, Journal of Biochemistry 142 (2007) 65-72.

A. M. Azevedo, J. M. S. Cabral, D. M. F. Prazeres, T. D. Gibson, L. P. Fonseca, Thermal and operational stabilities of Hansenula polymorpha alcohol oxidase, Journal of Molecular Catalysis B: Enzymatic 27 (2004) 37-45.


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