Application of Antioxidant Enzyme Activity and Biochemical Characterization in Static and Suspension Cultures of Withania somnifera L. towards Food Technology
食品生物技术的应用,
卷 2 编号 2 (2015),
20 三月 2015
,
第 15-22 页
https://doi.org/10.22037/afb.v2i2.7483
摘要
Withania somnifera (L.) Dunal is an erect evergreen shrub commonly known as Ashwagandha. It is widely used in Ayurvedic, and in the traditional pharmacopeia system of India. It is one of the major ingredients in many formulations prescribed for a variety of musculo-skeletal conditions including arthritis and rheumatism. In the present study, variations in the quality and quantity of proteins and antioxidant enzymes were evaluated biochemically and enzymatically from the static and suspension cultures of Withania somnifera L. The nodal segments provided the maximum callusing of 90.25±0.06% with (1mg/l) BAP and (2mg/l) Kn of 2, 4-D. The static and suspension cultures were taken for the analysis of total soluble protein, and screened for antioxidant enzyme activity [catalase (CAT), superoxide dismutase (SOD) and guaiacol peroxidase (GPX)]. The protein content (1.2016 μg/μl) was found to be higher in static culture samples (0.870 μg/μl) than the protein obtained from the suspension culture. The antioxidant enzyme activity (CAT, SOD and GPX) was higher in the static culture samples (301.01± 0.42, 198.92 ± 0.29, 103.75 ± 0.11 nkat/mg of protein) than in the suspension culture. Specific activity staining of isoenzyme pattern exhibited three isoforms (CAT 1, CAT 2 and CAT 3) in the static culture samples but CAT 1 was absent in the samples extracted from the suspension cultures. In case of SOD, four bands (SOD 1, SOD 2, SOD 3 and SOD 4) were found in both samples whereas the intensity of GPX activity was found to be more in the static culture; however, both samples exhibited three isoforms (GPX 1, GPX 2 and GPX 3). Supplementation of the required nutrients along with phytohormones under in vitro conditions might be an enhancing factor to yield antioxidant enzymes in the static culture samples.- Antioxidant
- Catalase
- Isoforms
- Protein Withania somnifera
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参考
- Fazly BS, Khajeh-karamadin M, Shokoohei-zadeh HR. In vitro antibacterial activity of Rheum ribes extract obtained from various plant parts against clinical isolates of Gram-negative pathogens. Iranian J Pharm Res. 2005; 2: 87-91.
- Kanungo S, Pradhan C, Sahoo SL, Sahu RK. Role of auxin in the in vitro rooting and micropropagation of Holarrhena antidysenterica Wall, a woody aromatic medicinal plant, through nodal explants from mature trees. J Med Plants Res. 2012; 6(31): 4660-4666.
- Mirjalili MH, Moyano E, Bonfill M, Cusido RM, Palazon J. Steroidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules.2009; 14 (7):2373–2393.
- Allan E: Plant cell culture. In: Stafford, A., Warren, G. (eds), Plant Cell and Tissue Culture. Chichester: John Wiley and Sons, 1996: 1-23.
- Phillips GC, Hubstenberger JF, Hansen EE. Plant regeneration by organogenesis from callus and cell suspension cultures. In: Gamborg, O.L., Phillips, G.C. (eds), Plant Cell, Tissue and Organ Culture. Heidelberg: Springer and Verlag, 1995: 67-78.
- Kanungo S, Sahoo SL. Direct organogenesis of Withania somnifera L. from apical bud. Int Res J Biotechnol. 2011; 2(3): 58-61.
- Valko M, Leibfritz D, Moncol J, Cronin M, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol. 2007; 39: 44–84.
- Sies, Helmut. Oxidative stress: Oxidants and antioxidants. Exp physiol.1997;82:291–295.
- Christen Y. Oxidative stress and Alzheimer disease. Am J ClinNutr.2000; 71:621-629.
- Nunomura A, Castellani R, Zhu X, Moreira P, Perry G, Smith M. Involvement of oxidative stress in Alzheimer’s disease. J NeuropatholExp Neurol. 2006; 65:631–641.
- Wood-Kaczmar A, Gandhi S, Wood N. Understanding the molecular causes of Parkinson's disease. Trends Mol Med.2006; 12:521–528.
- Hitchon C, El-Gabalawy H. Oxidation in rheumatoid arthritis. Arthritis Res Therapy.2004; 6: 265–278.
- Cookson M, Shaw P. Oxidative stress and motor neuron disease. Brain Pathol.1999; 9:165– 186.
- Nakabeppu Y, Sakumi K, Sakamoto K, Tsuchimoto D, Tsuzuki T, Nakatsu Y. Mutagenesis and carcinogenesis caused by the oxidation of nucleic acids. Biol Chem. 2006; 387: 373–379.
- Stadtman E. Protein oxidation and aging.Science.1992; 257:1220–1224.
- Maiani G, Periago CMJ, Catasta G, Carotenoids: Actual knowledge on food sources, intakes, stability and bioavailability and their protective role in humans. MolNutr Food Res. 2008; 53:194–218.
- Boskou D, Blekas G, Tsimidou M. Phenolic compounds in olive and olives. Curr Top Nutraceuti Res. 2005; 3:125–136.
- Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures.Physiol Plant. 1962; 15: 473-497.
- Moscatiello R, Baldan B, Navazio L. Plant Cell Suspension Cultures. In: Frans, J.M.M. (eds). Plant Mineral Nutrients: Methods and Protocols, Methods in Molecular Biology. Springer Science and Business Media, 2013: 84.
- Moon A, Moon N. Investigation of the antibacterial activity of Psoralea corylifolia leaf and callus extracts against multidrug resistant periodontal disease causing microorganisms. Int J Life Sci Pharma Res. 2012; 2(2):122-127.
- Misra BB, Dey S. Phytochemical analyses and evaluation of antioxidant efficacy on in vitro callus extract of east India Sandalwood tree (Santalum album L.). J Pharma Phytochem. 2012; 1(3):7-16.
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin-Phenol reagents.J Biol Chem. 1951; 193:265-275.
- Aebi H: Catalase, In: Packer L (eds.). Methods in enzymology. Academic press, Orlando, 1984: 121-126.
- Woodbury W, Spencer AK, Stahmann MA. An improved procedure using ferricyanide for detecting catalseisozymes. Anal Biochem.1971; 44: 301-305.
- Das K, Samanta L, Chainy GBN.A modified spectrophotometric assay of superoxide dismutase using nitrite formation by superoxide radicals. Ind J Biochem Biophys.2000; 37: 201-204.
- Beauchamp C, Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem.1971; 44:276–287.
- Bergmeyer HU: Methods of enzymatic analysis, Second Ed. Academic press, New York, 1974: 495.
- Hamill DE, Brewbaker JL. Isoenzyme polymorphism in flowering plants. IV. The peroxidiseisoenzymes of maize (Zea mays L.). Physiol Plant.1969; 22:945-958.
- Raha S, Roy SC. Efficient Plant Regeneration in Holarrhena antidysenterica Wall. from Shoot Segment-Derived Callus. In Vitro Cell Dev Biol Plant. 2003; 39(2): 151-155.
- Bhattacharya R, Bhattacharya S. High frequency in vitro propagation of Phyllanthus amarus Schum& Thom by shoot tip culture. Indian J Exp Biol. 2001; 39:1184-1187.
- Marks TR, Simpson SE. Factors affecting shoot development in apically dominant Acer cultivars in vitro. J Hort Sci. 1994; 60:543-551.
- Preece JC, Sutter EG. Acclimatization of micropropagation plants to the greenhouse and the fields. In: Debergh, P.C., Zimmerman, R.H. (ed.): Micropropagation. Kluwer Academic Publishers, Dordrecht, 1991: 71-93.
- Asaka ILI, Hirotani M, Asada Y, Furuya T. Production of ginsenosidesaponins by culturing ginseng (Panax ginseng) embryonic tissue in bioreactors. Biotechnol Lett.1993; 15: 1259-1264.
- Junaid A, Mujid A, Fatima S, Sharma MP. Cultural conditions affect somatic embryogenesis in Catharanthus roseusL. (G.) Don. Plant Biotech Rep. 2008; 8: 60-69.
- Lukas AM, Christoper DG, Rebecca AS, Virgnia W. AN9- a petunia gluthine s-transferase required for anthocyanin sequestration, is a flavonoid binding protein. Plant Physiol. 2000; 123:1561-1570.
- Ogita S. Callus and suspension culture of bamboo plant, Phyllostachys niger. Plant Biotech. 2005; 22(2): 119-125.
- Fujimura T, Komamine A. Synchronization of Somatic Embryogenesis in a Carrot Cell Suspension Culture. Plant Physiol. 1979; 64:162-164.
- Hussein EA, Taj-Eldeen AM, Al-Zubairi AS, Elhakimi AS, Al-Dubaie AR. Phytochemical screening, total phenolics and antioxidant and antibacterial activities of callus from Brassica nigra L. Hypocotyl explants. Int J Pharm. 2010; 6(4):464-471.
- Nathiya S, Santhi S, Senthil K. A comparative study on ontogenic expression of antioxidants and secondary metabolites in Withania somnifera. Int. Res. J. pharm.2012; 3(1):210-216.
- Kanungo S, Rout JR, Sahoo SL. Evaluation of antioxidant enzyme activities in Withania somnifera L. in vitro and in vivo grown explants. Iran J Biotechnol. 2013; 11 (4): 260-264.
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