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Int J Cur Sci Res. 2011; 1(3):134- 136

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International Journal of Current Scientific Research

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Short Report

Amino Acid Composition of a Mushroom Lentinus tuberregium VKJM24 (Hm060586)

J Manjunathan *a , V Kaviyarasanb

*a b

Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai-600 025 Assistant Professor, Centre for Advanced Studies in Botany, University of Madras, Chennai- 600 025.

ARTICLE INFO Keywords: Lentinus tuberregium, Aminoacid HPL

ABSTRACT

A total of 20 amino acids were recorded in Lentinus tuberregium and detected by HPLC analysis. However, the maximum amounts of aspartic acid (2.08 g), glutamic acid (1.87 g), isoleucine (1.12 g) were recorded. whereas in mycelium aspartic acid (0.86 g), glutamic acid (1.78 g), isoleucine (1.98 g) were recorded. From these studies, it was concluded that the supplementation of this mushroom with cereal diet would help to overcome lysine deficiency. The present study proved the potential of mushrooms which can enhance the health status of an individual

c Copyright 2011. CurrentSciDirect Publications. IJCSR - All rights reserved.

1. Introduction Edible mushrooms have been recognized from time immemorial. During World War II, yeasts were used extensively as food and fodder supplement in Germany and Jamaica [1]. Torula species from waste by-products have been used for human consumption [2-4]. Attention has been paid to scientific cultivation of mushrooms since the 17th century, and it is reported that mushrooms are being used extensively in many countries for food and fodder [5-7]. Mushrooms have better flavor and taste, and the same nutritive value as do Torula species [8]. They possess extensive enzyme complexes which eniable them to flourish successfully on a wide variety of iniexpensive substrates, such as lignin, cellulose, hemicelluloses, pectin, and other industrial wastes which are not suitable even for animal feed [9]. Mushrooms represent one of the world's greatest untapped resources of nutritious and palatable foods. Studies on the nutritive value and composition of a few species of mushrooms, and recent investigations by Lintzel [10], Fitzpatrick, Esselen, and Weir [11] have shown that in addition to the flavoring properties the proteins of some mushrooms are equal to muscle protein in nutritive value. Other recent investigations have shown the amino acid composition of some important edible mushrooms [12,13].

* Corresponding Author : J.Manjunathan Centre for Advanced Studies in Botany University of Madras, Guindy Campus, Chennai-600 025 Email:[email protected] c Copyright 2011. CurrentSciDirect Publications. IJCSR - All rights reserved.

According to Robinson and Davidson [14], the efficiency of protein production, from a given quantity of carbohydrates, in mushrooms and other higher fungi is about 65 % compared with about 20 % for pork, 15 % for milk, 5 % for poultry, and 4 % for beef. Lentinus grows abundantly on dead and decaying wood in clusters, and is eaten by the majority of people in India. This fungus is able to compete successfully with other fungi on undecomposed paddy straw and can be cultivated at a wide range of temperatures (21 to 33 C, relative humidity 67 to 72 %; Bano and Srivastava, [15] unlike field mushrooms which need exact conditions for their cultivation. The ease with which this mushroom can be cultivated economically on a large scale, within a period of 3 weeks, offers the possibility of its use in dietetics. With this object in view, it was of interest to find out the amino acid composition of Lentinus tuberregium. This paper deals with the qualitative and quantitative analysis of amino acids of this 2. Materials and Methods 2.1 Cultivation of mushrooms Lentinus tuberregium was grown on paddy straw beds prepared from paddy straw soaked in water for 15 hr. The size of the paddy straw beds might vary, but the best results were achieved in beds of 1 ft2 and 9 in. in thickness. The beds were kept on a raised platform under shade. Spawns of Lentinus tuberregium was prepared by inoculating sterilized paddy straw in a bag; 1- monthold spawns were used for inoculating the beds. Cajanus cajans (red gram) powder (40 mesh) was the best source of nutrient in the beds. The beds were watered twice a day, and the mushrooms

Manjunathan & Kaviyarasan / Int J Cur Sci Res. 2011; 1(3):134- 136

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appeared 20 days after inoculation. The yield of mushrooms was about 150 to 200 g per bed (Bano and Srivastava, 1963). Fresh mushrooms were taken and dried in a desiccator (over P205) to constant weight. Samples for analysis were prepared as described below. 2.2. Estimation of amino acid The amino acid composition was determined by highperformance liquid chromatograph (HPLC) based amino acid analyzer attached with fluorescence detector. The standard mixed chromatograms were established such as aspartic acid, glutamic acid, isoleucine, threonine, methionine, cystine, lysine, asparagines, glycine, arginine, valine, tryptophan, tyrosine, serine, leucine, phenylanine, histidine, alanine, gulatamine and proline. The test solution was prepared by dissolving the substance which was examined in the mobile phase for obtaining a concentration of 1.0 mg/ml. For reference solution, mixed amino acids Control Reference Standard (CRS) were dissolved in the mobile phase for obtaining a concentration of 1.0 mg/ml. The column was prepared by octadecylsilyl silica gel for chromatography R (3 µm) which acts as stationary phase. The size of the column should be l = 0.10 m, Ø = 4.6 mm. The stock solutions of 20µl of test solution and standard solution of mixed standard aminoacids were prepared by dissolving in double distilled water and then the mixture was constituted by mixing 1 mL each of the 21 standard amino acid solution and this was later used to establish the standard chromatogram. For the mobile phase, 15.2g of triethylamine R was dissolved in 800 ml of distilled water and the pH was adjusted to 3.0 with phosphoric acid R and final volume was make-up to 1000 ml with distilled water. From this 850 ml of the solution was added to a mixture of 2 volumes of propanol R and 3 volumes of acetonitrile R. The free amino acids in the standard and in L.tuberregium were automatically derivate by reacting with o-phthaldialdehyde under basic conditions to produce o-phthaldialdehyde derivatives in the reaction columns of the amino acid analyser. Two derivative reagent solutions were prepared as follows: 10 mL of 0.01 M sodium borate (Na2B4O7.10H2O) buffer solution B (pH 9.1) were added to 10 mL of b-mercaptopropionic acid to make reagent solution I. Reagent solution II was prepared by mixing 10 mL of 0.01 M sodium borate (Na2B4O7.10H2O) buffer solution B (pH 9.1) with 10 mg of o-phthaldialdehyde (OPA) dissolved in 3 mL of ethanol. Solutions I and II were filtered through 0.45 mm membrane filter before use. Following derivatization, the buffer solution A (mixed in acetonitrile in a 2:1 v/v ratio), containing the derivatized amino acid was transferred into the narrow bore HPLC system (HPLC column SRT ODSM, internal diameter = 4.6 and length = 150 mm) for separation at a temperature of 45°C with 20 µL injection volume and a flow rate volume of 1.0-1.5 mL/min. The detection was done using spectrophotometer at 220nm and the run time was about 90 min. 3. Results and Discussion A total of 20 amino acids were recorded in L. tuberregium and detected by HPLC analysis. However, the maximum amounts of aspartic acid (2.08 g), glutamic acid (1.87 g), isoleucine (1.12 g) were recorded [12]. The amino acid content varied in mushroom

species reported the amino acid contents in A. bisporus and P. ostreatus, they contained most of the amino acids. L. tuberregium contained all the essential amino acids; among which, aspartic acid (2.08 g), glutamic acid (1.87 g), were the major components [14]. The maximum level of vitamins such as niacinamide (10.65 mg/100g), folic acid (2.40 mg/100g), was recorded. The results of the present study clearly revealed that cultivation of L. tuberregium is simple, inexpensive and competitive to L. edodes. Temperature 20-25°C favored good yield of L. tuberregium. Table1. Aminoacid Composition In Fruitbody Of Lentinus Tuberregium Parameters Aminoacid Composition In Fruitbody Of Lentinus Tuberregium (100gm) 2.089 gms 1.87 gms 1.121 gms 1.087 gms 1.076 gms 1.044 gms 1.023 gms 0.997 gms Asparagine Glycine Arginine Valine Tryptophan Tyrosine Serine Leucine Phenylanine Histidine Alanine Gulatamine Proline 0.987 gms 0.9743 gms 0.7856 gms 0.7044 gms 0.643 gms 0.454 gms 0.4434 gms 0.404 gms 0.344 gms 0.221 gms 0.1121 gms In traces

Aspartic acid Glutamic acid Isoleucine Threonine Methionine Cystine lysine

Fig-1.Aminoacid composition in Lentinus tuberregium (Fruitbody)

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Manjunathan & Kaviyarasan / Int J Cur Sci Res. 2011; 1(3):134- 136

Fig-1.Aminoacid composition in Lentinus tuberregium (Fruitbody)

[11] Esselen W B, Fellers CR. Mushrooms for food and flavor. Mass. Agr. Exptl. Sta. Bull. No. 1946; 434. [12] Seelkopf C, Schuster H Qualitative and quantitative Amino Saurebestimmungen an einigen Wichtigen Speisepilzen. Z. Untersuch. Lebensm. 1957;106:177-187. [13] Reusser F, T. Spencer JF, Salans HR. Protein and fat content of some mushrooms grown in submerged culture. Appl Microbiol.1958; 6:1-4. [14] Robinson RF, Davidson RS. The large-scale growth of higher fungi. Advan. Appl. Microbiol.1959; 1:261-278. [15] Bano Z, Srivastava HC. Cultivation of Pleurotus species on paddy straw. Food Sci. 1962;11:363-365.

Mushroom as compared with fruits and vegetables is a better source of protein, containing lysine, arginine, histidine, and threonine in high concentrations. The essential amino acid composition of protein shows that mushroom is primarily deficient in phenylalanine and methionine, when compared with egg protein (Block and Mitchell, 1946) At the same time, when compared with the proportions of essential amino acids required for satisfactory mammalian growth, using tryptophan level as unity, the amino acid pattern of the mushroom protein appears to be adequate in all other amino acids, except phenylalanine and methionine. Supplementation of mushroom protein with phenylalanine and methionine would be necessary, when used as a sole source of protein in diet, to promote adequate growth. The composition of protein of this mushroom is approximately similar to that of Agaricus campestris except for the tryptophan content, which is higher in Pleurotus species [15]. This mushroom is being utilized by people in different areas, and has been found to be nontoxic. Since mushrooms are considered as delicacies, their supplementation with a cereal diet may help to overcome lysine deficiency. Further work on the biological value and protein efficiency ratio might throw more light on the nutritive value of the protein. The present study proved the potential of mushrooms which can enhance the health status of an individual. 4. References

[1] Prescott SC, Dunn CD. Industrial microbiology, 2nd ed. McGraw-Hill Book Co., Inc., New York, 1949. [2] Reiser C 0. Torula yeast from potato starch wastes. J Agr Food Chem.1954; 2:70-74. [3] Wiley AJ, Dubey GA, Lueck BF, Hughes LP. Torula yeast grown on spent sulfite liquor. Ind Eng Chem. 1950;42:1830-1833. [4] Wiley A J, Holderby JM, Fries KW. Food and feed yeast in the U.S.A., p. 89100. Proc. Tech. Panel Wood Chem. Food Agr. Organ. U.N., Stockholm, 1953. [5] Botticher W, Pannwitz N. The utilization of mushrooms growing in the f o r e s t s o f G e r m a n y a s f o o d a n d f o d d e r. Vo r r a t s p f l e g e Lebensmittelforsch.1941; 4:488-497. [6] Anderson, E. E., AND C. R. Fellers. 1942. Food value of mushrooms (Agaricus campestris). Proc. Am. Soc. Hort. Sci. 41-.301-304. [7] Gilbert F A, Robinson RF. Food and fungi. Econ Botany.1957; 11:126-145. [8] Block S S, Stearns TW, Stephens RL, Mc-Candless RFJ. Mushroom mycelium, experiments with submerged culture. J Agr Food Chem. 1953; 1:890-893. [9] Waksman SA. Decomposition of cellulose and hemicellu loses by microorganisms, p. 828-852. In Louis E. Wise [ed.], Wood chemistry. Reinhold Publishing Corp., New York, 1944. [10] Lintzel W. Uber den Nahrwert des Eiweisses der Speisepilze. Biochem J. 1941;308:413-419.

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