CHARACTERIZATION OF PHENYLALANINE AMMONIA LYASE PURIFIED FROM LOCALLY CULTIVATED GRAPE SEEDS (Vitis vinifera L.)
DOI:
https://doi.org/10.36103/ijas.v54i2.1727Keywords:
PAL, optimal condition, enzyme kinetics.Abstract
This study was conducted to investigate certain characteristics of the purified Phenylalanine ammonia lyase (PAL). The results revealed that the molecular weight was 180 kDa by gel filtration and Native-PAGE. The optimal pH and temperature for enzyme activity were 7 and 40 °C respectively. The enzyme activity was stable against pH values range of 6-8. The enzyme retained 79% of its total activity after incubation for one hour at 50 °C and retained 7% (lost 93%) of its activity at 70 °C. The entire activity was completely lost when the enzyme was incubated for 1 hour at temperatures over 70 °C. Carbohydrate content of the purified enzyme was estimated to be 24.61%. Km, Kcat and Vmax were estimated with phenylalanine as substrate, the values were 0.021 mM, 20.67 sec-1 and 4.13 mM/min respectively.
References
Al-Atrushy, S. 2021. Effect of foliar application of zinc and salicylic acid on vegetative growth and yield characteristics of halawani grape cultivar (Vitis vinifera L.). Iraqi Journal of Agricultural Sciences, 52(4): 989-998. https://doi.org/10.36103/ijas.v52i4.1410
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
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
Calabrese, J. C., D. B. Jordan, A. Boodhoo, S. Sariaslani and T. Vannelli. 2004. Crystal structure of phenylalanine ammonia lyase: multiple helix dipoles implicated in catalysis. Biochemistry, 43:11403-11416.
Castro, A. H. F., , M. C. Coimbra, S. T. D. Da Fonseca and A. A. De Melo souza. 2020. Phenylalanine ammonia-lyase in higher plants: Akey enzyme for plant development. Advances In Chemistry Research, 77: 62587- 62603.
Cochrane, F.C., L.B. Davin, N.G. Lewis. 2004. The Arabidopsis phenylalanine ammonia lyase gene family: kinetic characterization of the four PAL isoforms. Phytochemistry 65: 1557–1564.
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.
Dubois, M., Hamilton, J. and Smith, F. 1956. Colorimetric method for determination of sugars and related substances. Anal. Biochem., 28: 350-356
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
Emes, A. and L. Vining. 1970. Partial purification and properties of L-phenylalanine ammonia-lyase from Streptomyces verticillatus. Canadian Journal of biochemistry, 48, 613-622.
Fritz, R. R., D. Hodgins and C. Abell. 1976. Phenylalanine ammonia-lyase. Induction and purification from yeast and clearance in mammals. Journal of Biological Chemistry, 251, 4646-4650
Given, N., M. Venis and D. Grierson. 1988. Purification and properties of phenylalanine ammonia-lyase from strawberry fruit and its synthesis during ripening. Journal of Plant Physiology, 133: 31-37.
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. and C. H. Scaman. 2013. Purification and characterization of phenylalanine ammonia lyase from Trichosporon cutaneum. Enzyme research, 6: 1-7
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
Hadeel Y. and A. Khalida. Optimum condition for phenylalanine ammonia lyase extraction and purification of from locally cultivated grape seeds. Iraqi Journal Of Agricultural Sciences, (under publication(
Hao, Z., D. J. Charles, L. Yu and J. E. Simon. 1996. Purification and characterization of a phenylalanine ammonia-lyase from Ocimum basilicum. Phytochemistry, 43: 735-739.
Hyun, M. W., Y. H. Yun, J. Y. Kim and S. H. Kim. 2011. Fungal and plant phenylalanine ammonia-lyase. Mycobiology, 39: 257-265.
Hyun, M. W., Y. H. Yun, J. Y. Kim and S. H. Kim. 2011. Fungal and plant phenylalanine ammonia-lyase. Mycobiology, 39: 257-265.
Jorrin, J., R. Lopez-valbuena and M. Tena.1988. Purification and properties of phenylalanine ammonia-lyase from sunflower (Helianthus annuus L.) hypocotyls. Biochimica et Biophysica Acta (BBA)-General Subjects, 964: 73-82.
Kalghatgi, K. and P. Subba rao. 1975. Microbial L-phenylalanine ammonia-lyase. Purification, subunit structure and kinetic properties of the enzyme from Rhizoctonia solani. Biochemical Journal, 149: 65-75.
Kim, S. H., J. W . Kronstad and B. E. Ellis. 1996. Purification and characterization of phenylalanine ammonia-lyase from Ustilago maydis. Phytochemistry, 43: 351-357.
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.
Kovacik, J. and B. Klejdus. 2012. Tissue and method specificities of phenylalanine ammonia-lyase assay. Journal of Plant Physiology, 169: 1317-1320.
Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685
Lim, H.-W., S.-S. Park and C.-J. Lim. 1997. Purification and properties of phenylalanine ammonia-lyase from leaf mustard. Molecules and Cells (Springer Science & Business Media BV), 7: 158-178
Martin, M. E., E. Grao-cruces, M. C. Millan-linares and S. Montserrat-de la paz. 2020. Grape (Vitis vinifera L.) seed oil: a functional food from the winemaking industry. Foods, 9: 1360
Moffitt, M. C., G. V. Louie, M. E. Bowman, J. Pence, J. P. Noel and B. S. Moore. 2007. Discovery of two cyanobacterial phenylalanine ammonia lyases: kinetic and structural characterization. Biochemistry, 46: 1004-1012.
Nagai, N., Y. Kojima, M. Shimosaka and M. Okazaki. 1988. Effects of kinetin on L-phenylalanine ammonia-lyase activity in tobacco cell culture. Agricultural and biological chemistry, 52: 2617-2619.
Pridham, J. and S. Woodhead. 1974. Multimolecular forms of phenylalanine-ammonia lyase in Alternaria. Portland Press Ltd, 1974: 1070-1072
Reichert, A.I., X.Z. He, R.A. Dixon. 2009. Phenylalanine ammonia-lyase (PAL) from tobacco (Nicotiana tabacum): characterization of the four tobacco PAL genes and active heterotetrameric enzymes1. Biochem. J. 424: 233–242
Rahmatabadi, S. S., I. Sadeghian, , Y. Ghasemi, A. Sakhteman and S. Hemmati. 2019. Identification and characterization of a sterically robust phenylalanine ammonia-lyase among 481 natural isoforms through association of in silico and in vitro studies. Enzyme Microb Technol, 122: 36-54
Segel, I.H. 1976. Biochemical Calculations. John Wiely and Sons, New York, London, 280: 232-235
Shi, R., C.M. Shuford, J.P. Wang, Y.H. Sun, Z. Yang, H.C. Chen, S. Tunlaya-Anukit, Q. Li, J. Liu, D.C. Muddiman, R.R. Sederoff, V.L. Chiang. 2013. Regulation of phenylalanine ammonia-lyase (PAL) gene family in wood forming tissue of Populus Trichocarpa. Planta 238: 487–497.
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: 103861
Varga, A., P. Csuka, O. Sonesouphap, G. Banoczi, M. I. Tosa, G. Katona, Z. Molnar, L. C. Bencze, L. Poppe and C. Paizs. 2021. A novel phenylalanine ammonia-lyase from Pseudozyma antarctica for stereoselective biotransformations of unnatural amino acids. Catalysis Today, 366: 185-194
Zhu, L., W. Cui, Y. Fang, Y. Liu, X. Gao and Z. Zhou. 2013. Cloning, expression and characterization of phenylalanine ammonia-lyase from Rhodotorula glutinis. Biotechnology Letters, 35: 751-756.
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