The Identification of B-Ocimene Biosynthetic Pathway Through Mevalonate Acid (MVA) and 1-Deoxy-D-Xylulose 5-Phosphate (DXP) Pathways Using Crude Enzyme Extracts in Indonesian Bay Leaf/Salam Leaf (Syzygium polyanthum)
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Abstract
Salam leaf has a ?-ocimene as a key volatile compound that gives a fresh aroma to the food when the salam leaves are involved in the cooking process. As a secondary metabolic product, enzymatic biosynthesis as the early stage of ?-ocimene is a factor that needs to be known. Thus, this study was done to identify the mechanism of the two well-known terpenoid biosynthetic pathways, namely MVA and DXP pathways, in the biosynthesis of ?-ocimene in salam leaves. The activity of the 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR)-MVA pathway-determining enzyme and 1-deoxy-Dxylulose- 5-phosphate synthase (DXS)-DXP pathway-determining enzyme in the crude enzyme and their derivative products of salam leaves were analysed for their changes by differences of substrate ratios and enzyme inhibitors. The results showed that the activity of the HMGR enzyme was lower significantly than the DXS enzyme based on the addition of variations to the substrate ratio. These results were also supported by the enzyme and substrate reaction products, MVA and IPP intermediates from the MVA pathway, which were significantly lower when compared to DXP and IPP intermediates from the DXP pathway. As the end product of the reaction, ?-ocimene gave a significantly higher value of the DXP pathway than the MVA pathway. Therefore, it can conclude that the mechanism of the biosynthetic pathway of ?-ocimene in salam leaves was synthesised via the DXP pathway. The production of ?-ocimene could have crosstalk-pathway through the MVA pathway, especially when the DXP pathway was blocked.
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References
Baidoo E E K, Xiao Y, Dehesh K and Keasling J D. (2014). Metabolite profiling of plastidial deoxyxylulose-5-phosphate pathway intermediates by liquid chromatography and mass spectrometry. In Methods in molecular biology, volume 1153. Springer, 57–76. https://doi.org/10.1007/978-1-4939-0606-2_5
Brammer L A, Smith J M, Wades H and Meyers C F. (2011). 1-Deoxy-D-xylulose 5-phosphate synthase catalyzes a novel random sequential mechanism. Journal of Biological Chemistry 286(42): 36522–36231. https://doi.org/10.1074/jbc.M111.259747
Carretero-Paulet L, Cairó A, Botella-Pavía P, Besumbes O, Campos N, Boronat A and Rodríguez-Concepción M. (2006). Enhanced flux through the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing deoxyxylulose 5-phosphate reductoisomerase. Plant Molecular Biology 62: 683–695. https://doi.org/10.1007/s11103-006-9051-9/
Cock I E and Cheesman M. (2018). Plants of the genus Syzygium (Myrtaceae): A review on ethnobotany, medicinal properties and phytochemistry. In Goyal M R and Ayeleso A O, eds. Bioactive Compounds of Medicinal Plants: Properties and Potential for Human Health. New York: Apple Academic Press, 35–84.
Eganathan P, Saranya J, Sujanapal P and Parida A. (2012). Essential oil composition of leaves of Syzygium makul Gaertn. Journal of Essential Oil Bearing Plants 15(5): 827–831. https://doi.org/10.1080/0972060X.2012.10644127
Hampel D, Mosandl A and Wüst M. (2005). Biosynthesis of mono- and sesquiterpenes in carrot roots and leaves (Daucus carota L.): Metabolic cross talk of cytosolic mevalonate and plastidial methylerythritol phosphate pathways. Phytochemistry 66(3): 305–311. https://doi.org/10.1016/j.phytochem.2004.12.010
Hartanti L, Yonas S M K, Mustamu J J, Wijaya S, Setiawan H K and Soegianto L. (2019). Influence of extraction methods of bay leaves (Syzygium polyanthum) on antioxidant and HMG-CoA Reductase inhibitory activity. Heliyon 5(4): E01485. https://doi.org/10.1016/j.heliyon.2019.e01485
Hidayat Y, Hermawati E, Setiasih S, Hudiyono S and Saepudin E. (2018). Antibacterial activity test of the partially purified bromelain from pineapple core extract (Ananas comosus [L.] Merr) by fractionation using ammonium sulfate acetone. AIP Conference Proceedings, 2023 (1): 020067. https://doi.org/10.1063/1.5064064
Hirata H, Ohnishi T and Watanabe N. (2016). Biosynthesis of floral scent 2-phenylethanol in rose flowers. Bioscience, Biotechnology and Biochemistry 80(10): 1865–1873. https://doi.org/10.1080/09168451.2016.1191333
Kang Z W, Liu F H, Zhang Z F, Tian H G and Liu T X. (2018). Volatile ?-ocimene can regulate developmental performance of peach aphid myzus persicae through activation of defense responses in chinese cabbage brassica pekinensis. Frontiers in Plant Science 9: 708. https://doi.org/10.3389/fpls.2018.00708
Keilwagen J, Lehnert H, Berner T, Budahn H, Nothnagel T, Ulrich D and Dunemann F. (2017). The terpene synthase gene family of carrot (Daucus carota L.): Identification of QTLs and candidate genes associated with terpenoid volatile compounds. Frontiers in Plant Science 8: 1930. https://doi.org/10.3389/fpls.2017.01930
Kirby J, Nishimoto M, Chow R W N, Baidoo E E K, Wang G, Martin J, Schackwitz W, Chan R, Fortman J L and Keasling J D. (2015). Enhancing Terpene yield from sugars via novel routes to 1-deoxy-D-xylulose 5-phosphate. Applied and Environmental Microbiology 81(1): 130–138. https://doi.org/10.1128/AEM.02920-14
Kuzuyama T and Seto H. (2012). Two distinct pathways for essential metabolic precursors for isoprenoid biosynthesis. Proceedings of the Japan Academy, Series B 88(3): 41–52. https://doi.org/10.2183/pjab.88.41
Miziorko H M. (2011). Enzymes of the mevalonate pathway of isoprenoid biosynthesis. Archives of Biochemistry and Biophysics 505(2): 131–143. https://doi.org/10.1016/j.abb.2010.09.028
Öztekin S and Martinov M. (2013). Medicinal and aromatic crop drying. Stewart Postharvest Review 9(2): 1–5. https://doi.org/10.2212/spr.2013.2.3
Rehman R, Hanif M A, Mushtaq Z and Al-Sadi A M. (2016). Biosynthesis of essential oils in aromatic plants: A review. Food Reviews International 32(2): 117–160. https://doi.org/10.1080/87559129.2015.1057841
Rothman S C, Helm T R and Poulter C D. (2007). Kinetic and spectroscopic characterization of type ii isopentenyl diphosphate isomerase from Thermus thermophilus: Evidence for formation of substrate-induced flavin species. Biochemistry 46(18): 5437–5445. https://doi.org/10.1021/bi0616347
Sharma S N, Jha Z, Sinha R K and Geda A K. (2015). Jasmonate-induced biosynthesis of andrographolide in Andrographis paniculata. Physiologica Plantarum 153(2): 221–229. https://doi.org/10.1111/ppl.12252
Skorupinska-Tudek K, Wojcik J and Swiezewska E. (2008). Polyisoprenoid alcohols: Recent results of structural studies. The Chemical Record 8(1): 33-45. https://doi.org/10.1002/tcr.20137
Suvarchala Reddy N V L, Aveti S, Anjum M and Ganga Raju M. (2015). Anti-hyperlipidemic activity of methanolic extract of syzygium alternifolium bark against high-fat diet and dexamethasone-induced hyperlipidemia in rats. Asian Journal of Pharmaceutical and Clinical Research 8(6): 165–168.
Tritsch D, Hemmerlin A, Bach T J and Rohmer M. (2010). Plant isoprenoid biosynthesis via the MEP pathway: In vivo IPP/DMAPP ratio produced by (E)-4-hydroxy-3- methylbut-2-enyl diphosphate reductase in tobacco BY-2 cell cultures. FEBS Letters 584(1): 129–134. https://doi.org/10.1016/j.febslet.2009.11.010
Vrhovsek U, Lotti C, Masuero D, Carlin S, Weingart G and Mattivi F. (2014). Quantitative metabolic profiling of grape, apple and raspberry volatile compounds (VOCs) using a GC/MS/MS method. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences 966: 132–134. https://doi.org/10.1016/j.jchromb.2014.01.009
Wei H, Xu C, Movahedi A, Sun W, Li D and Zhuge Q. (2019). Characterization and function of 3-Hydroxy-3-Methylglutaryl-CoA reductase in Populus trichocarpa: Overexpression of PtHMGR enhances terpenoids in transgenic poplar. Frontiers of Plant Science 10: 1476. https://doi.org/10.3389/fpls.2019.01476
Wright L P and Phillips M A. (2014). Measuring the activity of 1-deoxy-D-xylulose 5-phosphate synthase, the first enzyme in the MEP pathway, in plant extracts. In Rodríguez-Concepción M. (Ed.), Plant Isoprenoids. Methods in Molecular Biology (Methods and Protocols), vol. 1153. New York, NY: Humana Press, 9–20. https://doi.org/10.1007/978-1-4939-0606-2_2
Yuwono S S and Faustina D R. (2019). Effect of withering time and chopping size on properties of pucuk merah (Syzygium oleana) herbal tea. IOP Conference Series: Earth and Environmental Science 230: 012047. https://doi.org/10.1088/1755-1315/230/1/012047
Zhou Y J, Gao W, Rong Q, Jin G, Chu H, Liu W, Yang W, Zhu Z, Li G, Zhu G, Huang L and Zhao Z K. (2012). Modular pathway engineering of diterpenoid synthases and the mevalonic acid pathway for miltiradiene production. Journal of Americal Chemical Society 134(6): 3234–3241. https://doi.org/10.1021/ja2114486