Identification, Cloning and Structural Analysis of Major Genes from Portulaca oleracea L. Hairy Roots that Involved in the Biosynthesis of Dopamine

Authors

1 Department of Biotechnology, College of Agriculture, Bu-Ali Sina University, Hamadan, Iran

2 Department of Medicinal Plant Production, Nahavand University, Nahavand, Iran

3 Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

Abstract

Dopamine is one of the important medications of Portulaca oleracea L. To optimize the production of dopamine, one of the methods is the identification and engineering of metabolite pathways. To investigate the tyrosine decarboxylase (TDC) and tyrosinase, which seem to be the most important genes in dopamine synthesis pathway, hairy roots were produced from Portulaca oleracea using Agrobacterium rhizogenes and total RNA was extracted from hairy roots. A cDNA library was synthesized using RT-PCR. Then, the twogenes were amplified, isolated and cloned in a pTG 19-T vector. Bioinformatics' databases were used to predict the details of the structural, functional and biological characteristic of these genes. Nucleotide sequence analysis revealed that the cloned cDNAs expressed TDC and tyrosinase, and contained a single open reading frame of 1800 bp and 1750 bp, respectively. TDC has the most similarity with TDC of Arabidopsis thaliana (L.) Heynh.,but tyrosinase has 98% similarity withtyrosinase of Agaricus bisporus. Because of More negatively charged amino acids the TDC has hydrophobic properties, therefore affinity and hydrophilic chromatography can be used for purification of TDC. But tyrosinase has hydrophilic properties and hydrophobicity chromatography can be used for its purification. There were two peroxisomal signal peptide (KLAKEFEQL) and (KIEGRPLHL) in the TDC and tyrosinase, respectively. Therefore, they are biologically active in the peroxisomes, and included in biosynthesis dopamine through the transformation of L-lysine to L-dopa and finally to the dopamine. In conclusion, increasing the expression of TDC and tyrosinase through the genetic engineering can increase dopamine production in the Portolaca.

Keywords

References

1. Li JWH, Vederas JC. Drug discovery and natural products: end of an era or an endless frontier? Sci. 2009; 325:161–5.
2. Harvey AL. Natural products in drug discovery. Drug Discov Today. 2008;13:894–901.
3. Bensaddek L, Villarreal ML and Fliniaux MA. Induction and Growth of Hairy Roots for the Production of Medicinal Compounds. Electronic J. Integrative Bio Sci.2008;1:2-9.
4. Julsing KM, Quax WJ and Kayser O. The Engineering of Medicinal Plants: Prospects and Limitations of Medicinal Plant Biotechnology. Medicinal Plant Biotechnology.Wiley-VCH Verlag GmbH. 2008;1-8.
5. Cragg GM and Newman DJ. Plants as Source of Anticancer Agents. J Ethnophar., 2005; 100: 72-79.
6. Oliveira I, Valentão P, Lopes R, Andrade PB, Bento A and Pereira JA.“Phytochemical characterization and radical scavenging activity of Portulaca oleraceae L. leaves and stems,” Microchem J, 2009; 92:129–134.
7. Dweck AC. Purslane (Portulaca oleracea)-the global panacea".Personal Care Magazine. 2001;2:7-15.
8. Zhang C, Yan Q, Cheuk WK and Wu J. Enhancement of tanshinone production in Salvia miltiorrhiza hairy root culture by Ag+ elicitation and nutrient feeding. Planta Medica. 2004;70:147-151.
9. Akhonzadeh S. Encyclopedia of Iranian Medicinal Plants. Arjomand Press: Tehran, Iran. (in Persian). 2000.
10. El-Sayed MIK. Effects of Portulaca oleracea L. seeds in treatment of type-2 diabetes mellitus patients as adjunctive and alternative therapy. J Ethnopharmacol. 2011;137:643-651.
11. Moghadam YA, Piri KH, Bahramnejad B and Habibi P. Methyl Jasmonate and Salicylic acid effects on the dopamine production in hairy cultures of Portulaca oleracea (purslan). Bull. Env. Pharmacol. Life Sci.  2013;2: 89-94
12. Chen J, Shi YP and Liu JY. Determination of noradrenaline and dopamine in Chinese herbal extracts from Portulaca oleracea L. by high-performance liquid chromatography". J Chromatogr A. 2003;1003:127-132.
13. Daviesa KM and Espley RV. Opportunities and challenges for metabolic engineering of secondary metabolite pathways for improved human health characters in fruit and vegetable crops. New Zealand J Crop Hortic Sci. 2013;41:154-177.
14. Bhayat SI, Gowda HM, Eisenhut M. Should dopamine be the first line inotrope in the treatment of neonatal hypotension? Review of the evidence. World J Clinical Pediatrics. 2016;5:212–22.
15. Zhang K and Ni Y. Tyrosine decarboxylase from Lactobacillus brevis: soluble expression and characterization. Protein Expres. Purif. 2014;94:33–39.
16. Murashig T and Skoog F. a revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiology plantarum. 1962; 15.
17. Saify Nabiabad H, Yaghoobi MM, Jalali M and Hosseinkhani S. Expression Analysis and purification of human recombinant tissue type plasminogen activator (rt-PA) from transgenic tobacco plants. Prep Biochem Biotechnol. 2011;41:2:175-186
18. Babashpour S, Aminzadeh S, Farrokhi N, Karkhane A, Haghbeen K. Characterization of a chitinase (Chit62) from Serratia marcescens B4A and its efficacy as a bioshield against plant fungal pathogens. Biochemical Genetics. 2012; 50:722-735
19. Stephen Stoker H. Organic and Biological Chemistry. Cengage Learning. 2015; p. 371. ISBN 978-1-305-68645-
20. Haynie DT, Xue B. Superdomain in the protein structure hierarchy: the case of PTP-C2. Protein Sci. 2015;24: 874-82.
21. Allison R, Robert C, Christodoulou C, Vendruscolo M and Dobson M. A relationship between the transient structure in the monomeric state and the aggregation propensities of α-synuclein and β-synuclein. Biochem. 2014;53:7170–7183.
22. Shirazi Z, Piri Kh., Mirzaie Asl A. and Hasanloo T. Glycyrrhizin and Isoliquiritigenin Production by Hairy Root Culture of Glycyrriz glabra. J Med Plants Res. 2012; 31: 4640-4646.
23. De Backer D, Aldecoa C, Njimi H, Vincent JL.Dopamine versus norepinephrine in the treatment of septic shock: a meta-analysis. Critical Care Medicine. 2012; 40:725–30.
24. Ahmadi Moghadam Y, Piri Kh, Bahramnejad B, Ghiasvand T. Dopamine Production in Hairy Root Cultures of Portulaca oleracea (Purslane) Using Agrobacterium rhizogenes. J Agr Sci Tech. 2014;16: 409-420.
Journal of Medicinal plants and By-product
Volume 7, Issue 2
September 2018
Pages 153-162

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