Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera

Authors

  • Zetty Amirah Zulkifli Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor-Bahru, Malaysia
  • Zaidah Rahmat 1) Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor-Bahru, Malaysia; 2) Chemical Management Unit, University Laboratory Management Centre (PPMU),Universiti Teknologi Malaysia, 81310 Johor-Bahru, Malaysia; 3) Institute of Bioproduct Development, Universiti Teknologi Malaysia, 81310 Johor-Bahru, Malaysia

DOI:

https://doi.org/10.14719/pst.2020.7.2.730

Keywords:

protein profiling, 1D SDS-PAGE

Abstract

Moringa oleifera is widely known as multipurpose tree since all of its parts confer multiple functions. The leaf is highly favourable among consumers while the petiole is mostly wasted. There are numerous studies on the flavonoid and antioxidant property of the stem and twig. However, study on the petiole has never been done. There-upon, this study was conducted to develop protein profiling of the petiole. In this study, 6 different protein extraction methods were tested on the fresh petiole before its protein quantity and quality were checked via Bradford assay and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) respectively. The in-solution digestion was then done prior to LC-MS/MS analysis. The protein electrophoretic pattern from the SDS-PAGE proves that method 6 using Tris HCl buffer with incorporation of dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF) confers the best quality of protein. It produced the highest number of visible individual bands compared to other methods. Meanwhile, 93 proteins were successfully identified via LCMS analysis where the protein, signal response and carbohydrate metabolism categories confer the highest percentage. High quality and content of the protein extracted from the petiole including the antioxidant, anticancer and antidiabetic protein identified suggested that consuming this part of the plant could enhance nutrients of human body.

Downloads

Download data is not yet available.

Author Biography

Zaidah Rahmat, 1) Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor-Bahru, Malaysia; 2) Chemical Management Unit, University Laboratory Management Centre (PPMU),Universiti Teknologi Malaysia, 81310 Johor-Bahru, Malaysia; 3) Institute of Bioproduct Development, Universiti Teknologi Malaysia, 81310 Johor-Bahru, Malaysia

Education:

  • PhD (Plant Molecular Biology & Proteomics), Imperial College London, UK (2012)
  • MSc (Plant Biotechnology), Universiti Putra Malaysia, Malaysia (2001)
  • BSc (Hons Biotechnology), Universiti Putra Malaysia, Malaysia (1999)

Google scholar ID: http://scholar.google.com/citations?user=Xz7Ki5YAAAAJ&hl=en

References

1. Anwar F, Bhanger M. Analytical characterization of Moringa oleifera seed oil grown in temperate regions of Pakistan. J Agric Food Chem 2003;51(22):6558-63. https://doi.org/10.1021/jf0209894

2. Amaglo NK, Bennett RN, Curto RBL, Rosa EA, Turco VL, Giuffrida A. et al. Profiling selected phytochemicals and nutrients in different tissues of the multipurpose tree Moringa oleifera L., grown in Ghana. Food Chem 2010;122(4):1047-54. https://doi.org/10.1016/j.foodchem.2010.03.073

3. Dolly J, Rai PK, Amit K, Shikha M, Geeta W. Effect of Moringa oleifera Lam. leaves aqueous extract therapy on hyperglycemic rats. J Ethnopharmacol 2009;123(3):392-96.https://doi.org/10.1016/j.jep.2009.03.036

4. Kamath N, Swaminathan R, Desai N. Antibacterial activity of Indian medicinal plant- Moringa oleifera against MRSA and Klebsiella Spp. (ESBL) which are commonly isolated bacteria in hospital environments. Int J Adv Res. 2016;2(8):515-17

5. Torres C, Sanchez J, Osorio H, Sinagawa G, Aguirre A, Gutiérrez D. Moringa oleifera: Phytochemical detection, antioxidants, enzymes and antifugal properties. Int J Exp Bot. 2013;82:193-202

6. Ilyas M, Arshad M, Saeed F, Iqbal M. Antioxidant potential and nutritional comparison of moringa leaf and seed powders and their tea infusions. J Anim Plant Sci. 2015;25:226-33

7. Kumbhare MR, Guleha V, Sivakumar T. Estimation of total phenolic content, cytotoxicity and in–vitro antioxidant activity of stem bark of Moringa oleifera. Asian Pac J Trop Dis. 2012; 2(2):144-50. https://doi.org/10.1016/S2222-1808(12)60033-4

8. Atawodi SE, Atawodi JC, Idakwo GA, Pfundstein B, Haubner R, Wurtele G, Owen RW. Evaluation of the polyphenol content and antioxidant properties of methanol extracts of the leaves, stem and root barks of Moringa oleifera Lam. J Med Food. 2010;13(3):710-16. https://doi.org/10.1089/jmf.2009.0057

9. Sun LW, Ma PT, Li DN, Lei XJ, Ma R, Qi C. Protein extraction from the stem of Panax ginseng CA Meyer: a tissue of lower protein extraction efficiency for proteomic analysis. Afr J Biotechnol. 2011;10(21):4328-33.

10. Azri W, Ennajah A, Jing M. Comparative study of six methods of protein extraction for two-dimensional gel electrophoresis of proteomic profiling in poplar stems. Can J Plant Sci. 2013; 93:895-901. https://doi.org/10.4141/cjps2013-113

11. Ni M, Dehesh K, Tepperman JM, Quail PH. GT-2: in vivo transcriptional activation activity and definition of novel twin DNA binding domains with reciprocal target sequence selectivity. Plant Cell. 1996;8(6):1041-59. https://doi.org/10.1105/tpc.8.6.1041

12. Westermeier R, Naven T, Höpker HR. Proteomics in Practice: A Guide to Succesful Experimental Design. John Wiley & Sons 2008;2:67. https://doi.org/10.1002/9783527622290

13. Maranho RC, Benez MM, Maranho GB, de Azevedo Fernandes VN, Gonela A, Mangolin CA. da Silva Machado MFP. Extraction of total protein from axillary buds of sugarcane (Saccharum spp.) for proteomic Analysis. Sugar Tech. 2018;20(1):95-99. https://doi.org/10.1007/s12355-017-0520-z

14. Carpentier SC, Witters E, Laukens K, Deckers P, Swennen R, Panis B. Preparation of protein extracts from recalcitrant plant tissues: An evaluation of different methods for two?dimensional gel electrophoresis analysis. Proteomics. 2005;5: 2497-07. https://doi.org/10.1002/pmic.200401222

15. Horecker BL. The pentose phosphate pathway. J Biol Chem. 2002;277(50):47965-71. https://doi.org/10.1074/jbc.X200007200

16. Caverzan A, Passaia G, Rosa SB, Ribeiro CW. Lazzarotto F, Margis-Pinheiro M. Plant responses to stresses: role of ascorbate peroxidase in the antioxidant protection. Genet Mol Biol. 2012;35(4):1011-19. https://doi.org/10.1590/S1415-47572012000600016

17. Battin EE, Brumaghim JL. Antioxidant activity of sulfur and selenium: a review of reactive oxygen species scavenging, glutathione peroxidase and metal-binding antioxidant mechanisms. Cell Biochem Biophys. 2009;55(1):1-23. https://doi.org/10.1007/s12013-009-9054-7

Published

19-04-2020

How to Cite

1.
Zulkifli ZA, Rahmat Z. Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera. Plant Sci. Today [Internet]. 2020 Apr. 19 [cited 2024 Nov. 4];7(2):214-8. Available from: https://horizonepublishing.com/journals/index.php/PST/article/view/730

Issue

Section

Research Articles