Gallic acid is a phenolic compound ubiquitous in every part of the plant. It is quite high in Melissa officinalis plants and also known as lemon balm that belongs to Lamiaceae family. It is a short rooted and spreading herb which has anti-inflammatory, psychoneurological, antispasmolytic, antifungal, antiviral, antiparasitic and antioxidant activities. Medicinal drugs made from these leaves and plants have a faint lemon aroma that contains several phytochemicals and pharmacological properties making this herb an ideal for body and skin care use. Our aim is to, enhance gallic acid from the leaves of Melissa officinalis by Bacillus substilis through submerged fermentation. Our research showed that there is an increase of gallic acid upto 17.0 µg/ml during bio-transformation processes and bioanalytical techniques where conversion of tannic acid to gallic acid is triggered by tannase enzyme produced by Bacillus substilis. It was extracted from solvent extraction method from this plant and estimated by Folin–Ciocalteus reagent and was used in fermentation process for enhancement of gallic acid. The gallic acid concentration was increased and found to be 17.0µg/ml. Especially the gallic acid is enhanced due the activity of tannase enzyme which may converts tannic acid to gallic acid. This enzyme was produced by Bacillus substilis which acts as biocatalyst and converts tannic acid to gallic acid. Thus, this microorganism is essential for the biotransformation of tannic acid to gallic acid. In this research work, biotransformation has played a vital role for gallic acid enhancement. Glucose solution is consumed by Bacillus substilis. Here glucose is estimated by DNS reagent (3,5-Dinitrosalicylic acid) and tannic acid estimated by FD reagent (Folin – Denis reagent). Based on various time intervals, the concentration of gallic acid, enzyme concentration and biomass were increased but the tannic acid and glucose concentrations were decreased.

REFERENCES 

1. Shabby AS, El-Gengaihi S, Khattab M. Oil of Melissa     officinalis L., as Affected by Storage and Herb Drying. J Essent Oil Res. 1995;7(6):667-9. doi: 10.1080/10412905.1995.9700525.
2. Meftahizade H, Lotfi M, Moradkhani H. Optimization of Micropropagation and establishment of cell suspension culture in Melissa officinalis L. Afr J Biotechnol.2010;9(28):4314-21. https://www.ajol.info/index.php/ajb/article/view/82650 
3. Janina MS. Melissa officinalis. Int J Aromather. 2003;10:132-9.
4. Ghasemi M, Jafarpour M, Forogh M. Effect of different drying methods on the quality and quantity of the essential oil of lemon balm. Int J Agric Crop Sci. 2013;6(9):501-4. Corpus ID: 29710195
5. De Sousa AC, Alviano DS, Blank AF, Alves PB, Alviano CS, Gattass CR. Melissa officinalis L. essential oil: antitumoral and antioxidantactivities. J Pharm Pharmacol.2004;56(5):67781.doi:10.1211/0022357023321, PMID 15142347.
6. Ulbricht C, Brendler T, Gruen wald J, Kligler B, Keifer D, Abrams TR, Woods J, Boon H, Kirkwood CD, Hackman DA, Basch E, Lafferty HJ, Natural Standard Research Colla boration. Lemon balm (Melissa officinalis L.): an evidence-based systematicreviewbytheNaturalStandardResearchCollaboration.Pharmacother.2005;5(4):71114.doi:10.1080/J157v05n04_08, PMID 16635970.
7. Belsinger S. Lemon balm, herb of the year 2007. J Herbs Health. 2007;11(6):26-31.
8. Dragland S, Senoo H, Wake K, Holte K, Blomhoff R. Several culinary and medicinal herbs are important sources of dietary antioxidants. J Nutr. 2003;133(5):1286-90. doi: 10.1093/jn/133.5.1286, PMID 12730411.
9. Kennedy DO, Scholey AB, Tildesley NT, Perry EK, Wesnes KA. Modulation of mood and cognitive performance following acute administration of Melissa officinalis (lemon balm). Pharmacol Biochem Behav. 2002;72(4):953-64. doi: 10.1016/s0091-3057(02)00777-3, PMID 12062586.
10. Wölbling RH, Leonhardt K. Local therapy of herpes simplex with dried extract from Melissa officinalis. Phytomedicine. 1994;1(1):25-31. doi: 10.1016/S0944-7113(11)80019-X, PMID 23195812.
11. Marongiu B, Porcedda S, Piras A, Rosa A, Deiana M, Dessì MA. Antioxidant activity of supercritical extract of Melissa officinalis subsp. officinalis and Melissa officinalis subsp. inodora. Phytother Res. 2004;18(10):789-92. doi: 10.1002/ptr.1549, PMID 15551397.
12. Mulkens A, Kapetanidis I. Flavonoids of the leaves of Melissa officinalis L. (Lamiaceae). Pharm Acta Helv. 1987;62(1):19-22. PMID 3562477.
13. Agata I, Kusakabe H, Hatano T, Nishibe S, Okuda T. Melitric acids A and B, new trimeric caffeic acid derivatives from Melissa officinalis. Chem Pharm Bull. 1993;41(9):1608-11. doi: 10.1248/cpb.41.1608.
14. Tekel J, Vaver kov S, Kova cicova J, Hola M, Havra nek E. A  screening method for the determination of the residues of uracil herbicides in Melissa officinal
15. isL.1994;49;899:901.https://pascalfrancis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3357354
16. Patora J, Klimek B. Flavonoids from lemon balm (Melissa officinalis). Acta Pol Pharm. 2002;59(2):139-43. PMID 12365606.
17. Mrlianová M, Tekel’ová D, Felklová M, Reinöhl V, Tóth J. The influence of the harvest cut height on the quality of the herbal drugs Melissae folium and Melissae herba. Planta Med. 2002;68(2):178-80. doi: 10.1055/s-2002-20247, PMID 11859476.
18. Barros L, Dueñas M, Dias MI, Sousa MJ, Santos-Buelga C, Ferreira IC. Phenolic profiles of cultivated, in vitro cultured and commercial samples of Melissa officinalis L. infusions. Food Chem. 2013;136(1):1-8. doi: 10.1016/j.foodchem.2012.07.107, PMID 23017385.
19. Wojdy?o A, Oszmia?ski J, Czemerys R. Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chem. 2007;105(3):940-9. doi: 10.1016/j.foodchem.2007.04.038.
20. Cases J, Ibarra A, Feuillère N, Roller M, Sukkar SG. Pilot trial of Melissa officinalis L. leaf extract in the treatment of volunteers suffering from mild-to-moderate anxiety disorders and sleep disturbances. Med J Nutrition Metab. 2011;4(3):211-8. doi: 10.1007/s12349-010-0045-4, PMID 22207903.
21. Nasri H, Rafieian-Kopaei M. Protective effects of herbal antioxidants on diabetic kidney disease. J Res Med Sci. 2014;19(1):82-3. PMID 24672573.
22. Zarei A, Changizi-Ashtiyani S, Taheri S, Hosseini N. A brief overview of the effects of Melissa officinalis L. extract on the function of various body organs. Zahedan J Res Med Sci. 2015;17(7). doi: 10.17795/zjrms1007.
23. Rafieian-Kopaei M, Baradaran A, Rafieian M. Oxidative stress and the paradoxical effects of antioxidants. J Res Med Sci. 2013;18(7):629. PMID 24516501.
24. Herodež ŠS, Hadolin M, Škerget M, Knez Ž. Solvent extraction study of antioxidants from Balm (Melissa officinalis L.) leaves. Food Chem. 2003;80(2):275-82. doi: 10.1016/S0308-8146(02)00382-5.
25. Martins EN, Pessano NT, Leal L, Roos DH, Folmer V, Puntel GO, Rocha JB, Aschner M, Ávila DS, Puntel RL. Protective effect of Melissa officinalis aqueous extract against Mn-induced oxidative stress in chronically exposed mice. Brain ResBull.2012;87(1):749.doi:10.1016/j.brainresbull.2011.10.003, PMID 22020131.
26. Ribeiro MA, Bernardo-Gil MG, Esquível MM. Melissa officinalis, L.: study of antioxidant activity in supercritical residues. J Supercrit Fluids. 2001;21(1):51-60.doi: 10.1016/S0896-8446(01)00078-X.
27. Shaygannia E, Bahmani M, Zamanzad B, Rafieian-Kopaei M. A review study on Punica granatum L. J Evid Based Complementary Altern Med. 2016;21(3):221-7. doi: 10.1177/2156587215598039, PMID 26232244.
28. Ince AE, Sahin S, Samna SG. Extraction of phenolic compounds from Melissausing microwave andultrasound. Turk J Agric For. 2013; 37:69-75. Doi: 10.3906/tar-1201-1