Actas de congresos
Studies On Bromelain Precipitation By Ethanol, Poly (ethylene Glycol) And Ammonium Sulphate
Chemical Engineering Transactions. Italian Association Of Chemical Engineering - Aidic, v. 24, n. , p. 979 - 984, 2011.
Brazil produces over than 58.000 hectares of pineapple, beeing the world leader in pineapple production. Aiming the recovery of agriculture residues (stalk and leaves), and fruit processing residues (stem and bark), several researches have been carried out for the recovery of pineapple's enzymes, such as the bromelain. These enzymes performs important role in proteolytic modulation at cellular matrix, in numerous physiologic process, including tissue morphogenesis, tissue repair, angiogenesis and tissue modulation, decreasing bruises, swelling, pain and healing time. This work aims the recovery on bromelain from pineapple stem, bark and leaves. Aqueous extract of bromelain was prepared through stem, bark and leaves processing in a common blender. Precipitation studies were performed under refrigeration (4 °C). Poly (ethylene glycol) precipitations were carried out in a single step, in various concentrations. Ethanol and ammonium sulphate precipitations were carried out stepwise, where several concentrations of ethanol and ammonium sulphate were added to perform a fractional precipitation. Results showed that bromelain was not precipitated by poly(ethylene glycol) at all. However, bromelain was recovered with 30-70% ethanol fraction, in which were achieved a purification factor of 2.07 fold, and yielded over than 98% of enzymatic activity. Precipitation with ammonium sulphate showed that bromelain was recovered at 20-40 saturation fractions, yielding up to 44% of protein recovery, and a purification factor of 4.4 fold. Although the ethanol precipitation had achieved a lower purification factor, it seems to be more suitable for bromelain recovery, based on process time and local ethanol price, since Brazil is the largest ethanol producer worldwide. © 2011, AIDIC Servizi S.r.l.24979984Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding (1976) Analytical Biochemistry., 72, pp. 248-254Cesar, A.C., Silva, R., Lucarini, A.C., Recuperação das enzimas presentes na casca e no talo do abacaxi (1999) Revista de Iniciação Científica - Sao Carlos/SR, 1, pp. 47-53Devakate, R.V., Patil, V.V., Waje, S.S., Thorat, B.N., Purification and drying of bromelain (2009) Separation and Purification Technology, 64, pp. 259-264Englard, S., Seifter, S., Precipitation techniques (1990) Guide to Protein Purification, , Eds. Deutscher, M.P. Academic Press San Diego, USAFontes, L.V.Q., Campos, G.S., Beck, P.A., Brandao, C.F.L., Sardi, S.I., Precipitation of bovine rotavirus by polyethylen glycol (PEG) and its application to produce polyclonal and monoclonal antibodies (2005) Journal of Virological Methods, 123 (2), pp. 147-153. , DOI 10.1016/j.jviromet.2004.09.023, PII S0166093404002824Haq, S.K., Rasheedi, S., Sharma, P., Ahmad, B., Khan, R.H., Influence of salts and alcohols on the conformation of partially folded intermediate of stem bromelain at low pH (2005) International Journal of Biochemistry and Cell Biology, 37 (2), pp. 361-374. , DOI 10.1016/j.biocel.2004.07.005, PII S1357272504002626Harrach, T., Eckert, K., Maurer, H.R., Machleidt, I., Machleidt, W., Nuck, R., Isolation and characterization of two forms of an acidic bromelain stem proteinase (1998) Journal of Protein Chemistry, 17 (4), pp. 351-361. , DOI 10.1023/A:1022507316434Kumar, V., Sharma, V.K., Kalonia, D.S., Effect of polyols on polyethylene glycol (PEG)-induced precipitation of proteins: Impact on solubility, stability and conformation (2009) International Journal of Pharmaceutics, 366, pp. 38-43Maurer, H.R., Bromelain: Biochemistry, pharmacology and medical use (2001) Cellular and Molecular Life Sciences, 58 (9), pp. 1234-1245Narayan, A., Madhusudhan, M., Raghavarao, K., Extraction and purification of ipomoea peroxidase employing three-phase partitioning (2008) Applied Biochemistry and Biotechnology, 151, pp. 263-272Oliveira, L.A., Porto, A.L.F., Tambourgi, E.B., Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes (2006) Bioresource Technology, 97 (6), pp. 862-867. , DOI 10.1016/j.biortech.2005.04.017, PII S0960852405002233Rabelo, A.P.B., Tambourgi, E.B., Pessoa Jr., A., Bromelain partitioning in two-phase aqueous systems containing PEO-PPO-PEO block copolymers (2004) Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 807 (1), pp. 61-68. , DOI 10.1016/j.jchromb.2004.03.029, PII S1570023204002521Rowan, A.D., Buttle, D.J., Barrett, A.J., The cysteine proteinases of the pineapple plant (1990) Biochemical Journal, 266 (3), pp. 869-875Roy, I., Gupta, M.N., Current trends in affinity-based separations of proteins/enzymes (2000) Current Science., 78, pp. 587-591Silveira, E., Souza Jr., M.E., Santana, J.C.C., Chaves, A.C., Porto, A.L.F., Tambourgi, E.B., Expanded bed adsorption of bromelain (E.C. 220.127.116.11) from Ananas comosus crude extract (2009) Brazilian Journal of Chemical Engineering, 26, pp. 149-157Umesh Hebbar, H., Sumana, B., Raghavarao, K.S.M.S., Use of reverse micellar systems for the extraction and purification of bromelain from pineapple wastes (2008) Bioresource Technology, 99 (11), pp. 4896-4902. , DOI 10.1016/j.biortech.2007.09.038, PII S0960852407007845Walker, J.M., (2002) The Protein Protocols Handbook, , Humana Press, Totowa, New Jersey