OPTIMUM CONDITIONS FOR PHENYLALANINE AMMONIA LYASE EXTRACTION AND PURIFICATION FROM LOCALLY CULTIVATED GRAPE SEEDS
DOI:
https://doi.org/10.36103/ijas.v54i2.1728Keywords:
Phenylalanine ammonia lyase, grape seeds, optimization.Abstract
In the present study, Phenylalanine ammonia lyase (PAL) activity was detected in different parts of grape plant (leaves, pulp, whole seeds and defatted seeds) extracts. The results revealed that the whole seed possess highest PAL activity (69.44 U), followed by defatted seed (44.22 U), whereas extracts of leaves and pulp gave the lowest activity values (22.77 and 5.11 U, respectively). The optimum conditions for crude PAL extraction achieved using 50mM potassium phosphate buffer (pH 7) at mixing ratios of (1:7) for 1 h. extraction time. PAL was shown to precipitate out at the saturation range between 20-90%. For further purification, gel filtration chromatography using Sephadex G-200 column (1.5×70) and ion exchange chromatography using DEAE- Cellulose (DE 52) column (3.2× 10) were applied. The specific activity, number of folds and yield of purified PAL was 1157 U/mg, 19.94 and 15.32 % respectively.
References
Abood, S. C., and I. M. Hakeem, 2016. Purification and characterization of salivary amylase inhibitor extracted from barley. Iraqi Journal of Agricultural Science, 47(4): 1039-1048. https://doi.org/10.36103/ijas.v47i4.536
Adams, A. K., E. O. Wermuth and P. E. Mcbride. 1999. Antioxidant vitamins and the prevention of coronary heart disease. American family physician, 60, 895
Al-Atrushy, Sh. M. 2019. Effect of foliar application of micronutrients and canopy management on yield and quality of grapevine (Vitis vinfera L.) cv. Mirane. Iraqi Journal of Agricultural Sciences –1029:50(2):626-637. https://doi.org/10.36103/ijas.v2i50.662
Arafa, A., A. Abdel-Ghani, S. El-dahmy, S. Abdelaziz, Y. El-ayouty and A. El-sayed. 2020. Purification and Characterization of Anabaena flos-aquae Phenylalanine Ammonia-Lyase as a Novel Approach for Myristicin Biotransformation
Babaoglu Aydas, S., S. Ozturk and B. Aslim. 2013. Phenylalanine ammonia lyase (PAL) enzyme activity and antioxidant properties of some cyanobacteria isolates. Food Chem, 136, 164-9
Campos, R., H. Nonogaki, T. Suslow and M. E. Saltveit. 2004. Isolation and characterization of a wound inducible phenylalanine ammonia-lyase gene (LsPAL1) from Romaine lettuce leaves. Physiologia Plantarum, 121, 429-438
Carbajal, D., A. Casaco, L. Arruzazabala, R. Gonzalez and V. Fuentes. 1991. Pharmacological screening of plant decoctions commonly used in Cuban folk medicine. Journal of Ethnopharmacology, 33, 21-24
Cheng, G. W. and P. J. Breen. 1991. Activity of phenylalanine ammonia-lyase (PAL) and concentrations of anthocyanins and phenolics in developing strawberry fruit. Journal of the American Society for Horticultural Science, 116, 865-869
Cui, J. D., J. Q. Qiu, X. W. Fan, S. R. Jia and Z. L. Tan. 2014. Biotechnological production and applications of microbial phenylalanine ammonia lyase: a recent review. Critical reviews in biotechnology, 34, 258-268
Derradji-benmeziane, F., R. Djamai and Y. Cadot. 2014. Antioxidant capacity, total phenolic, carotenoid, and vitamin C contents of five table grape varieties from Algeria and their correlations. OENO One, 48, 153-162
Eissa, H. A. and W. A. Ibrahim. 2018. Kinetics of phenylalanine and tyrosine ammonia-lyase enzymes activity of banana fruit (Musa cavendishii L., cv. Enana). Middle East Journal of Applied Sciences, 8, 680-689
Goldson, A., M. Lam, C. H. Scaman, S. Clemens, and A. Kermode. 2008. Screening of phenylalanine ammonia lyase in plant tissues, and retention of activity during dehydration. Journal of the Science of Food and Agriculture, 88, 619-625
Goldson Barnaby, A., R. Reid and D. Warren. 2017. Phenylalanine ammonia lyase activity, antioxidant properties, fatty acid profile, mineral content and physiochemical analyses of Cissus sicyoides berries. Journal of Berry Research, 7, 117-127
Gonzalez-Mendoza, D., R. Troncoso-Rojas, T. Gonzalez-Soto, O. Grimaldo-Juarez, C. Cecena-Duran, D. Duran-Hernandez and F. Gutierrez-Miceli. 2018. Changes in the phenylalanine ammonia lyase activity, total phenolic compounds, and flavonoids in Prosopis glandulosa treated with cadmium and copper. An Acad Bras Cienc, 90, 1465-1472
Hajhashemi, V., G. Vaseghi, M. Pourfarzam and A. Abdollahi. 2010. Are antioxidants helpful for disease prevention? Research in pharmaceutical sciences, 5, 1
Hashim, S. T., and M. M. Nema, 2018. production and purification of chitinase from Aeromonas spp. and study its effect on some cancer cell lines in vitro. Iraqi Journal of Agricultural Science, 49(3): 438-444. https://doi.org/10.36103/ijas.v49i3.115
Hemmati, S. 2015. Phenylalanine ammonia-lyase through evolution: A bioinformatic approach. Trends in Pharmaceutical Sciences, 1, 10-14
Jungbauer, A. and R. Hahn. 2009. Ion-exchange chromatography. Methods in enzymology, 463, 349-371
Kim, D. S.and B. K. Hwang. 2014. An important role of the pepper phenylalanine ammonia-lyase gene (PAL1) in salicylic acid-dependent signalling of the defence response to microbial pathogens. J Exp Bot, 65, 2295-306
Kosyk, O. I., I. M. Khomenko, L. M. Batsmanova and N. Y. Taran. 2017. Phenylalanine ammonia-lyase activity and anthocyanin content in different varieties of lettuce under the cadmium influence. The Ukrainian Biochemical Journal, 89, 85-91
Koukol, J. and E. E. Conn, 1961. The metabolism of aromatic compounds in higher plants: IV. Purification and properties of the phenylalanine deaminase of Hordeum vulgare. Journal of Biological Chemistry, 236, 2692-2698
Kyriakidis, N. B. and T. Katsiloulis. 2000. Calculation of iodine value from measurements of fatty acid methyl esters of some oils: comparison with the relevant American oil chemist’s society method. Journal of the American Oil Chemists' Society, 77, 1235-1238
Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. nature, 227, 680-685
Levy, H. L., C. N. Sarkissian and C. R. Scriver. 2018. Phenylalanine ammonia lyase (PAL): From discovery to enzyme substitution therapy for phenylketonuria. Mol Genet Metab, 124, 223-229
Lim, H.-W., S.-S. Park and C.-J. Lim. 1997. Purification and properties of phenylalanine ammonia-lyase from leaf mustard. Molecules & Cells (Springer Science & Business Media BV), 7
Lister, C. E., J. E. Lancaster and J. R. Walker. 1996. Phenylalanine ammonia-lyase (PAL) activity and its relationship to anthocyanin and flavonoid levels in New Zealand-grown apple cultivars. Journal of the American Societr Horticultural Science, 121, 281-285
Machado, N. F. and R. Dominguez-perles. 2017. Addressing facts and gaps in the phenolics chemistry of winery by-products. Molecules, 22, 286
Mulero, J., G. Martinez, J. Oliva, S. Cermeno, J. Cayuela, P. Zafrilla, A. Martinez-Cacha and A. Barba. 2015. Phenolic compounds and antioxidant activity of red wine made from grapes treated with different fungicides. Food chemistry, 180, 25-31
Ofagain, C., P. M. Cummins and B. F. Oconnor, 2011. Gel-filtration chromatography. Protein Chromatography, 25-33
OIV, F. 2016. FAO-OIV Focus Table and Dried Grapes. Vol. I7042 FAO, 64, 440
Peterson, E. A. and H. A. Sober. 1962. Column chromatography of proteins: substituted celluloses. Methods in enzymology. Elsevier vol. 5, pp. 3-27
Rao, P. S., K. Moore and G. Towers. 1967. Degradation of aromatic amino acids by fungi: ii. purification and properties of phenylalanine ammonia-lyase from ustilago hordei. Canadian journal of biochemistry, 45, 1863-1872
Roca, P., L. Castro-Lopez, G. Castillo-Sanchez, L. Diaz-Rubio and I. Cordova-Guerrero. 2019. Total content of phenols and antioxidant activity of grape skins and seeds cabernet sauvignon cultivated in Valle de Guadalupe. Baja California, Mexico. BIO Web of Conferences 15
Shahidi, F. and Y. Zhong. 2010. Novel antioxidants in food quality preservation and health promotion. European Journal of Lipid Science and Technology, 112, 930-940
Shi, J., J. Yu, J. E. Pohorly and Y. Kakuda. 2003. Polyphenolics in grape seeds—biochemistry and functionality. Journal of Medicinal Food, 6, 291-299
Solekha, R., F. A. Susanto, , T. Joko, , T. R. Nuringtyas and Y. A. Purwestri. 2019. Phenylalanine ammonia lyase (PAL) contributes to the resistance of black rice against Xanthomonas oryzae pv. oryzae. Journal of Plant Pathology, 102, 359-365
Tang, G.-Y., C.-N. Zhao, Q. Liu, X.-L. Feng, X.-Y. XU, S.-Y. Cao, X. Meng, S. LI, R.-Y. Gan and H.-B. Li. 2018. Potential of grape wastes as a natural source of bioactive compounds. Molecules, 23, 2598
Unusan, N. 2020. Proanthocyanidins in grape seeds: An updated review of their health benefits and potential uses in the food industry. Journal of Functional Foods, 67
Whetten, R. W. and R. R. Sederoff. 1992. Phenylalanine ammonia-lyase from loblolly pine: purification of the enzyme and isolation of complementary DNA clones. Plant physiology, 98, 380-386.
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
2.jpg)
2.jpg)
