Extraction And Preliminary Characterization Of Bromelain From Curaua (ananas Erectifolius L.b.smith) Purple And White

Abstract

The characterization and determination of kinetic parameters of bromelain extracted from curaua (Ananas erectifolius) white and purple varieties were studied. Optimal pH and temperature conditions for proteolytic activity were determined by azocasein method. The enzyme has shown optimum activity at pH 6.0 for white and 6.0 and 7.0 for purple curaua, and at 40 °C and 60 °C for white curaua and at 40°C for the purple curaua. The lowest result of Km and the highest result of Vmax/Km were found for purple curaua, but the two varieties showed affinity by the substrate, with Km value of 185.18 μmolL-1min, Vmax of 158.73 μmolL-1 and activation energy of 13,903 for white curaua and Km of 192.0 μmolL-1min, Vmax of 188.68 μmolL-1 and activation energy of 14,941 cal/mol for purple curaua. This enzyme showed lower activation energy (26,00028,000 cal/mol) than those of other coexisting proteinases. Copyright © 2014,AIDIC Servizi S.r.l.37769774Akuzawa, R., Okitani, A., Purification and characterization of a cysteine proteinase with low activation energy from lactococcus lactis ssp. Lactis iam 1198 (2010) Journal of Dairy Science, 78, pp. 2609-2616Amid, A., Ismail, N., Yusof, F., Salleh, H., Expression, purification and characterization of a recombinant stem bromelain form ananas comosus (2011) Process Biochemistry, (46), pp. 2232-2239Blandio, A., Macias, M., Cantero, D., Immobilization of glucose oxidase within calcium alginate gel capsules (2001) Process Biochemistry, 36, pp. 601-606Bradford, M.M., (1976) Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding Analytical biochemistry, (72), pp. 248-254Corzo, C.A., Krzysztof, N.W., Chanes, J.W., Pineapple fruit bromelain affinity to different protein substrates (2012) Food Chemistry, 133 (3), pp. 631-635Ferreira, J.F., (2007) Characterization and purification of bromelain enzyme in aqueous two phase systems, , (in Portuguese), PhD Thesis, Faculdade de Engenharia Química, Universidade Estadual de Campinas, Campinas, BrasilFerreira, J.F., Rocha Antunes Pereira Bresolin, I., Silveira, E., Tambourgi, E.B., Purification of bromelain from ananas comosus by peg/phosphate atps (2011) Chemical Engeneering Transactions, 6 (24), pp. 931-936Fujihashi, G.A., Barbosa, W.L.R., Ananas erectifolius (curaua): Standardization of the extracts, fractions and plant material (2002) Scientific journal UFPA, 3, pp. 1-6Halpern, M.J., (1997) Biochemistry 1, pp. 233-303. , Ed. Lisbon, Ed. Lidel PortugalLeão, A.L., Rowell, R., Tavares, N., Aplications of natural fibers in automotive industry in brazil-Thermoforming process (1998) Science and Technology of Polymers and Advanced Materials-Emerging Technologies and Business Opportunities, , New YorkLeão, A.L., Caraschi, J.C., Tan, I.H., Curaua fiber-A tropical natural fibers from amazon potential and applications in composites (2000) Natural Polymers and Agrofibers Composites, , Sao Carlos, BrazilLineweaver, H., Burk, D., The detennination of enzyme dissociation constants (1934) J. Am. Chern. Soc, (56), p. 658Nishizawa, K., Shimura, K., Nyumon, K.K., (1972) An Introduction to Enzymatic Chemistry, , Konando, Tokyo, JapanOkino, N., Ikeda, R., Ito, M., Expression, purification, and characterization of a recombinant neutral ceramidase from mycobacterium tuberculosis (2010) Biosci. Biotechnol. Biochem, (4), pp. 316-321Oliveira, E.C.P., Lameira, O.A., Sousa, F.I.B., Silva, R.J.F., Leaf curaua structure at different intensities of photosynthetically active radiation (2008) Pesq. Agropec. Bras.,Brasília, 43 (2), pp. 163-169. , In PortugueseRowan, A.D., Buttle, D.J., Barrett, A.J., The cysteine proteinases of the pineapple plant (1990) Biochemical Journal, 266 (3), pp. 869-875Rowell, R.M., Han, J.S., Rowell, J.S., (2000) Natural Polymers and Agrofibers Based Composites, , IQSC/Embrapa Instrumentação Agropecuária/UNESP, São Carlo