Extraction and purification of papain obtained from three native species of the genus Vasconcellea
Published 2023-04-30
Keywords
- Vasconcellea,
- papaína,
- liofilización,
- purificación,
- actividad enzimática
How to Cite
Abstract
In this research, the papain enzyme was obtained from the latex of three species of the Vasconcellea genus, V. chachapoyensis, V. heilbornii and V. pubescens, species native to the Amazon region in Peru. The extracted latex was conditioned with sodium acetate buffer, EDTA and sodium bisulfite solution. The purification was carried out adapting the method of Andrade (2011), ammonium sulfate was used for precipitation; 96° ethanol in two stages for dilution; the pH was regulated at 6.0 and 8.0 with NaOH 0.1 N, it was centrifuged in three stages; frozen at -65 °C for 12 hours; and lyophilized from 24 to 36 hours with a vacuum of 0.1 to 0.6 mbar and a collector temperature of -84 to -85 ° C, finally it was packed in dark bottles. Using the milk coagulation method (Balls and Hoover) the enzymatic activity (AE) was determined, the three species obtained higher values of AE at pH 6.0 than at pH 8.0; V. chachapoyensis presented a higher significant average of EA (240.97 Upe), followed by V. pubescens (195.80 Upe) and V. heilbornii (112.45 Upe), it was also significantly higher than commercial papain (208.14 Upe). V. pubescens had higher proteolytic activity than the other species.
References
- Acosta, R. (2011). Estudio del variación de la actividad enzimática proteolítica del látex del Babaco (Vasconcellea heilbornii cv babaco) en función de la edad del fruto. Escuela Politécnica Nacional.
- Andrade, M., Morales, O., & Martínez, H. (2011). Estudo do processo de extração de papaína a partir do látex do fruto de mamão (Carica papaya L.) cv. Maradol. Acta Agronómica, 60(3 SE-), 218–224. https://revistas.unal.edu.co/index.php/acta_agronomica/article/view/28818
- Arana, P., & Quijano, M. (2012). Extracción, caracterización y comparación de latéx obtenido en secado por aspersión, de variedades de papaya (Carica papaya L.). In Escuela Superior Politécnica del Litoral. Escuela Superior Politécnica del Litoral.
- Arellano, J. V. (2019). Extracción de la enzima papaína presente en el chilacuan (Vasconcellea pubescens) como alternativa de cuajo vegetal [Universidad Politécnica Estatal del Carchi]. http://repositorio.upec.edu.ec/handle/123456789/877
- Gutiérrez, A., Nolasco, O., & Santa, C. (2017). Purification and preliminary characterization of latex proteases of Vasconcellea candicans (A. Gray) A. DC (Mito). Scientia Agropecuaria, 8(1), 7–17. https://doi.org/10.17268/sci.agropecu.2017.01.01
- Herrera, E., & Ruiz, Lord. (2014). Efecto del tiempo de congelación y temperatura de liofilizado del látex del fruto papayita de monte (Carica Pubescens) sobre su actividad proteolítica en leche de vaca [Universidad Nacional Toribio Rodriguez de Mendoza de Amazonas]. http://repositorio.untrm.edu.pe/handle/UNTRM/553
- Kyndt, T., Van Damme, E. J. M., Van Beeumen, J., & Gheysen, G. (2007). Purification and characterization of the cysteine proteinases in the latex of Vasconcellea spp. FEBS Journal, 274(2), 451–462. https://doi.org/10.1111/j.1742-4658.2006.05592.x
- Lin, N. M., Linn, K. S., Than, Y. M., & Thu, M. K. (2020). Enzymatic Studies on Crude Papain from Papaya Peels. Maubin University Research Journal, 11, 173–180.
- Muriel, O., Gutiérrez, P., Quequezana, M., & Arenas, C. (2005). Extracción de la papaína de Carica var. Arequipensis. Véritas, 9(1), 128–135.
- Ocampo, J. (2021). Papaya de Olor (Vasconcellea pubescens). Research Gate, March.
- Puig, A. M. (2008). Desarrollo de un proceso para la extracción de papaína en Colombia. In Universidad de los Andes. Universidad de los Andes.
- Ramirez, F. (2019). La papaína y su potencial como producto de valor agregado en Costa Rica. Research Gate, April. https://doi.org/10.13140/RG.2.2.32076.46729
- Schedelman, X., Kyndt, T., Ming, R., & Droogenbroeck, V. (2011). Wild Crop Relatives: Genomic and Breeding Resources. In Wild Crop Relatives: Genomic and Breeding Resources (Issue June). https://doi.org/10.1007/978-3-642-20447-0
- Sinche, M. (2009). Aislamiento, purificación parcial y caracterización cinética de las proteasas presentes en el látex de los frutos de una planta del género Vasconcella [Escuela Politecnica Nacional]. http://bibdigital.epn.edu.ec/handle/15000/1661
- Tacias-Pascacio, V. G., Morellon-Sterling, R., Castañeda-Valbuena, D., Berenguer-Murcia, Á., Kamli, M. R., Tavano, O., & Fernandez-Lafuente, R. (2021). Immobilization of papain: A review. International Journal of Biological Macromolecules, 188(October), 94–113. https://doi.org/10.1016/j.ijbiomac.2021.08.016
- Tineo, D., Bustamante, D. E., Calderon, M. S., Mendoza, J. E., Huaman, E., & Oliva, M. (2020). An integrative approach reveals five new species of highland papayas (Caricaceae, Vasconcellea) from northern Peru. PLoS ONE, 15(12 December). https://doi.org/10.1371/journal.pone.0242469
- Torres, M. J., Trejo, S. A., Martin, M. I., Natalucci, C. L., AviléS, F. X., & López, L. M. I. (2010). Purification and characterization of a cysteine endopeptidase from vasconcellea quercifolia A. St.-Hil. latex displaying high substrate specificity. Journal of Agricultural and Food Chemistry, 58(20), 11027–11035. https://doi.org/10.1021/jf904295x
- Tovar-Colmenárez, Y., Ávila de Hernández, R., Pire-Sierra, M. C., & Gonzáles-Ortíz, M. T. (2018). Purificación de la papaína del látex de la lechosa y cuantificación de la actividad enzimática. Revista Científica A.S.A, August, 44–59.