Journal of Medicinal Plants Research Vol. 3(11), pp. 897-900, November, 2009 Available online at http://www.academicjournals.

org/jmpr ISSN 1996-0875 2009 Academic Journals

Full Length Research Paper

Amino acid and GABA content in different cultivars of Momordica charantia L.

Yong Kyoung Kim1, Hui Xu1, Nam IL Park1, Hee Ock Boo2, Sook Young Lee3 and Sang Un Park1*
1

Division of Plant Science and Resources, Chungnam National University, 220 Gung-Dong, Yuseong-Gu, Daejeon, 305764, Korea. 2 Department of Biotechnology, Chosun University, 375 Seosuk-Dong, Dong-Gu, Gwangju, 501-759, Korea. 3 Research Center for Oral Disease Regulation of the Aged, Chosun University, 375 Seosuk-Dong, Dong-Gu, Gwangju, 501-759, Korea.
Accepted 24 August, 2009

The variability of amino acid levels including gamma-aminobutyric acid (GABA) was investigated in six cultivars of bitter melon (Momordica charantia L.) of different origins: Nikko and Peacock from Japan, Galaxy and Verde Buenas from Philippines and two native cultivars from China and Korea. Cultivars varied considerably in the amounts of different amino acids. Among them, the cultivar Verde Buenas recorded the highest amounts of seven different amino acids with the content of other amino acids at a satisfactory level. The cultivar Galaxy contained the highest amount of GABA. Second only to Verde Buenas, the Korean native performed the best, having significantly high levels of five different amino acids followed by the China native exhibiting the highest amounts of alanine, phenylalanine and proline. Although the cultivar Nikko contained the highest amount of arginine, it recorded the lowest values for most of the amino acids. The cultivars Nikko, Peacock and Galaxy did not contain methionine amino acids. These results demonstrated that bitter melon cultivars varied in the amount of different amino acids and GABA and hence have a high potential as a good source of amino acids and GABA in the development of food supplements. Key words: Amino acid, cultivar, gamma-aminobutyric acid, GABA, Momordica charantia L. INTRODUCTION Momordica charantia L. is a herbaceous vine belonging to family Cucurbitaceae. This crop is widely grown for its edible fruit, which is among the most bitter of all vegetables. All parts of the plant, including the fruit, taste bitter and hence commonly known as bitter melon. It is cultivated throughout the world to use as a vegetable as well as for its medicinal value. Fruits and seeds of bitter melon are known to have anti-HIV, anti-ulcer, antiinflammatory, anti-leukemic, antimicrobial, anti-diabetic and anti-tumor properties (Basch et al., 2003; Shih et al., 2008; Alam et al., 2009; Lii et al., 2009). Amino acids are organic acids containing both amine and carboxyl functional groups, having a variety of roles in metabolism. One of the most important functions of amino acids is to act as building blocks of proteins. A huge amount of different amino acids exist in nature, among them around two dozen of them are vital in human nutrition (Gilani et al., 2008; Millward et al., 2008). Gamma-aminobutyric acid (GABA, Figure 1) is a nonprotein amino acid synthesized via the decarboxylation of L-glutamate in a reaction catalyzed by the cytosolic enzyme L-glutamate decarboxylase (Forde and Lea, 2007). It is a valuable component of the free amino acid pool in most prokaryotic and eukaryotic organisms. In vertebrates, GABA is the major neurotransmitter inhibitor in the central nervous system that prevents anxiety and stress-related messages from reaching the motor centers of the brain (Schousboe and Waagepetersen, 2007). It has been used to enhance the secretion of growth hormone from the pituitary gland. In addition, it is known to inhibit the migration of colon carcinoma cells, paving the way to the development of specific pharmacological agents that delay or inhibit invasion and metastasis of

*Corresponding author. E-mail: supark@cnu.ac.kr. Tel: +82-42821-5730. Fax: +82-42-822-2631.

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Figure 1. Chemical structure aminobutyric acid (GABA). of

RESULTS From the analysis of different cultivars of bitter melon collected from Korean, China, Japan and Philippines, seventeen different amino acids were found to be in varying proportions (Table 1). The range of detected amino acid content varied widely irrespective of the place of origin and the type of cultivar tested. Among all the amino acids isolated, arginine was found to be in remarkably high quantities, while the limiting amino acids were cysteine and methionine. One of the important amino acids found in large amounts irrespective of cultivar type is arginine. The mean concentration of arginine was between 28.0 and 52.6 mol/g D.W., which was several times significantly higher than any other amino acid found in this study. The amount of each amino acid varied significantly within a particular cultivar. Among the cultivars of different locations, the cultivar Verde Buenas contained the highest amount of seven different amino acids, that is, aspartic acid, glycine, isoleucine, leucine, methionine, threonine and valine. Besides these seven amino acids, the same cultivar contained a satisfactory amount of other amino acids. The cultivar Galaxy contained the highest amount of glutamic acid (5.7 mol/g dry weight (D.W.)) although it contained the lowest amounts of arginine, aspartic acid, cysteine, lysine and tyrosine. The Korean native performed the best in terms of five different amino acids, that is, cysteine, histidine, methionine, serine and tyrosine whereas it contained the lowest amounts of Isoleucine and threonine. The Chinese native cultivars had the highest amount of alanine, phenylalanine and proline while arginine content was the lowest. The Japanese cultivar Nikko, although contained the highest amount of arginine (52.6 mol/g D.W.) recorded the lowest values for most of the other amino acids: cysteine, glutamic acid, glycine, histidine, leucine, phenylalanine, proline, serine and valine. Another Japanese cultivar Peacoak contained the highest amount of lysine (3.7 mol/g D.W.). There was no methionine in the cultivars, Nikko, Peacoak and Galaxy. The cultivars exhibited significant variability in their GABA content, varying between 4 - 20 mol/g D.W. (Figure 2). The Philippines cultivar Galaxy contained the highest amount (19.3 mol/g D.W.) of GABA followed by the Chinese native (14.0 mol/g D.W.) which was around five times more than the other cultivars. The cultivars Peacock, the Korean native, Verde Buenas and Nikko contained as low as 3.5, 4.2, 4.8 and 5.2 mol GABA /g dry weight, respectively. DISCUSSION Bitter melon contained seventeen amino acids at satisfactory levels, except cysteine and methionine. Among them, the amount of arginine (52.6 mol/g D.W.), alanine (21.7 mol/g D.W.) and GABA (19.3 mol/g D.W.) were quite high irrespective of the type of cultivar. As this

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various cancer types (Enna, 2001; Kowalski et al., 2007; Sarto-Jackson and Sieghart, 2008). There have been a number of comments as to the role that GABA might play in plants including, the involvement in pH regulation, nitrogen storage, plant development and defense and as a compatible osmolyte as well as an alternative pathway for glutamate utilization (Alan and Barry, 1997; Barry et al., 1999). Most of the research work of bitter melon was done considering its medicinal value but a few researches were done based on amino acid and GABA content. The objectives of this study were to conduct a comparative analysis of amino acid and GABA content of different cultivars of bitter melon form China, Japan, Philippines and Korea.
MATERIALS AND METHODS Plant material Fruits of six different cultivars of M. charantia: (one Korean native, one Chinese native, two Japanse cultivars (Nikko and Peacock from Japan) and two Philippines cultivars (Galaxy and Verde Buenas) were collected from the experimental farm of Chungnam National University (Daejeon, Korea) in September 2008. Fresh samples were stored frozen in sealed clear polyethylene plastic bags at -80C until used. Collected samples were freeze-dried at 80C for at least 72 h and dried samples were ground into a fine powder (40-mesh) by a mill. Free amino acid extraction Amino acids were extracted from freeze-dried plant powder (1 g) with 30 ml of 70% ethanol at 80oC for 20 min. After evaporating ethanol, residue water phase (30 ml) was mixed with ethyl ether (30 ml) in a separation funnel. The water phase was separated and freeze-dried. The extract was resuspended in 3 ml of 0.02 N HCl and filtered using a 0.45 m syringe filter. The extraction procedure was replicated three times. Analysis of amino acids The amino acid analyzer (HITACHI L-8900, Japan) attached HITACHI HPLC Packed Column with Ion-exchanging Resin No. 2622 PF (4.6 60 mm) and UV detector (VIS1: 570 nm, VIS2: 440 nm) was used for analysis of amino acids. Wako L-8500 buffer solution PF-1, 2, 3, 4 and RG were used in this study. Twenty l of each sample was injected and determination was performed using Ninhydrin reagent set (Wako Chemical Inc, Japan). All samples were run in triplicates.

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Table 1. Amino acid compositions in the fruit of different cultivars of M. charantia L. (The unit of amino acid content = mol/g dry weight).

Type of amino acid Alanine Arginine Aspartic acid Cysteine Glutamic acid Glycine Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Proline Serine Threonine Tyrosine Valine

Native (Korea) 4.7 0.6 28.4 2.1 3.9 0.5 0.5 0.1 3.4 0.34 1.7 0.17 7.6 0.76 2.4 0.20 2.5 0.27 3.0 0.29 0.3 0.29 3.1 0.41 2.9 1.46 4.9 0.48 2.6 0.68 7.0 0.60 6.8 0.18

Mean Amino Acid Content of Different Cultivars (mol/g D.W.) Native Nikko Peacoak Galaxy (China) (Japan) (Japan) (Philippines) 21.7 1.28 5.3 0.76 4.2 0.4 13.81.5 28.0 3.5 52.6 5.26 48.2 3.2 28.01.8 4.4 0.6 4.9 0.7 7.0 1.0 3.5 0.5 0.2 0.0 0.1 0.0 0.1 0.1 0.1 0.0 5.1 0.5 1.8 0.1 2.3 0.2 5.7 0.5 1.8 0.3 1.2 0.1 2.2 0.1 1.7 0.2 7.4 0.8 3.8 0.5 4.0 0.3 5.9 0.5 4.7 0.3 2.5 0.2 5.0 0.4 4.6 0.3 3.2 0.2 1.6 0.1 2.6 0.2 2.7 0.2 3.5 0.7 2.5 0.2 3.7 0.3 2.1 0.1 0.3 0.0 0.0 0.0 0.0 0.0 0.0 0.0 6.1 0.4 2.4 0.1 4.3 0.2 4.2 0.2 7.3 1.4 1.5 0.1 1.8 0.2 3.3 0.5 4.8 0.3 2.5 0.1 3.1 0.4 3.8 0.3 6.8 0.5 9.0 0.5 5.1 0.5 10.1 1.2 3.9 0.4 1.9 0.13 3.9 0.4 2.4 0.2 9.9 0.9 5. 4 0.4 7.9 1.1 8.4 1.2

Verde Buenas (Philippines) 8.5 1.06 48.1 6.01 7.1 1.0 0.3 0.0 2.6 0.3 3.3 0.4 7.0 0.8 6.4 0.8 3.8 0.4 3.6 0.4 0.3 0.0 6.0 0.7 3.0 0.3 4.8 0.6 12.6 1.5 3.1 0.3 11.2 1.4

Figure 2. GABA content in the fruit of different cultivars of M. charantia L. (The unit of GABA content = mol/g dry weight).

vegetables contained almost all of the amino acids in satisfactory amounts, especially with regard to arginine, alanine and GABA, which were in very high, this crop could be a very good source in the production of arginine, alanine, GABA and other amino acids found in high concentrations, on a commercial basis.

ACKNOWLEDGEMENTS This study was carried out with the support of Cooperative Research Program for Agricultural Science and Technology Development (Project No. 20070101033118), RDA, The Republic of Korea.

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