INTRODUCTION:
Hippocrates first mentioned mushrooms when he wrote about their medicinal value in 400 B.C. The first mention of mushroom cultivation, distinct from a chance appearance in the field, was in l652. 1,2 Unfortunately, they were described as excellent for “making into compresses for ripening boils” but not as good to eat. In l707, a French botanist wrote about mushrooms as “originating from a horse.” People have harvested mushrooms from the world for thousands of years for food and medicines.
Mushrooms are superior to many vegetables and beans in their nutritive values. 3,4,5 Fresh mushrooms contain about 88.5%water and 3.2%protein. But in dried mushrooms water content is low and protein content is high as 34-44%. The protein content is superior to that in most vegetables, beans, peas, fruits, fishes, goat’s meat, chicken.6,7 Mushrooms contain about 4.2-4.4% carbohydrates. Carbohydrate is very low compared to the beans and vegetables. Crude fiber content is as high as in vegetables. So it can be provide to diabetic patients.8 ,9 The fat content is less than 0.3%. It is less than that in fish, meat, egg and milk.
The fat is rich in ergosterol which is involved in biosynthesis of vitamin in human body. 10,11,12 Mushrooms contain high proportion of thiamine, riboflavin, niacin and ascorbic acid. 13 They are also rich in minerals such as Ca, Fe, Na and K. Daily intake of 100g dried mushrooms meets the need of proteins, vitamins and minerals for an adult man.14,15 As the nutritive value is high, FAO has recommended dried mushrooms as a source of protein in the human. 16
MATERIALS AND METHODS:
1. Collection of Mushroom:
Fresh Agaricus bisporus was purchased from a mushroom farm in Ongole (AP, India) showed in Fig 1. In addition, the samples were cleaned without washing, cut into slices, dried at room temperature for 15 days, and then stored for further analysis.
2. Determination of Moisture content:15
The fresh weight (one gram) of mushroom sample was taken in a china dish using chemical balance. These samples were then oven dried separately at 80°C for 48 h. The loss in weight obtained after drying was regarded as the moisture content.
3. Determination of Ash Value: 15
Two gram sample from the finely ground mushroom was placed in a crucible and converted into ash at 550 - 600°C for 5 h in a carbolite muffle furnace after which it was allowed to cool in a desiccators. The difference in the weight of the crucible without the sample before and after ashing was used to calculate the ash content. This was repeated till two consecutive weights were the same and the ash was almost white or greyish white in colour. Then total ash was calculated as:
4. Determination of Acid insoluble ash: 15
Acid soluble ash is the residue obtained after boiling the total ash with dilute HCL and igniting the remaining insoluble matter. This measure the amount of silica present, especially as sand and siliceous earth Determination of acid insoluble ash value. Using 25 ml of dilute HCL; wash the total ash from the dish used for total ash in to a 100ml beaker. Place wire gauge over it a Bunsen burner and boil for 5mins. Fiter through an ‘ash less’ filter paper; wash the residue twice with hot water. Ignite crucible in the flame, cool and weigh. Put the filter paper and residue together in to the crucible; heat gently until vapours cease to be evolved and then more strongly until all carbon has been removed. Cool in a desiccators. Weigh the residue and calculate acid in soluble ash of the crude drugs with reference to the air dried sample of the true drug.
|
100 X (Z – V)
Y Z ---- Weight of the powder + Weight of Crucible Dish V ---- Weight of the Ash + Weight of Crucible Dish Y --- Weight of The Drug |
5. Determination of Water soluble ash: 15
Difference between the weight of total ash and the residue after treatment of the total ash with water.
|
Water soluble ash=Weight of Total ash-Weight of ash after treating with water |
6. Determination of Sulphated ash: 15
Take one grams of substances in weighed crucible Ignite as before allow to cool and weigh. Repeated the operation until 2 successive weightings do not differ by more than 0.5mg, ignite gently at first until the substances is thoroughly charred. Cool, moisten the residue with one ml of sulphuric acid; heat gently until white fumes no longer evolved and ignite at 800±25º c until black particles has disappeared. Allow the crucible to cool, add a few drops of sulphuric acid and heat.
|
Sulphated ash = Weight of Total ash - Weight of ash after treating with Sulphuric acid |
7. Determination of Fat: 16
Crude fat was determined using Soxhlet extraction apparatus. Petroleum ether (boiling point equals 40 - 60°C) was added to a 2.0 g finely ground mushroom sample placed in the extraction apparatus. Extraction was carried out for 16 h after which the ether was evaporated to dryness. The amount of fat was obtained from the difference in the weight of the flask and after drying of the ether.
|
Determination = Weight of R.B. Flask--Weight of Empty of Fat With fat R.B. Flask |
8. Determination of Crude fibre: 16
Crude fibre was determined using a Hennenberg-Stohmann method. A 2 g mushroom species was boiled in antifoam solution (1-octanol) for 30 min. Pyrex glasses were used to filter the solution where the residues were thoroughly washed with boiling water (3 times) to remove hydrochloric acid. The Pyrex glasses containing the residues were dried at 100°C for five hours, cooled to room temperature and then weighed. The crucibles were then placed in a muffle furnace at 555°C for 5 h, cooled to room temperature and then reweighed to find the fibre content percentage.
9. Determination of total carbohydrate:17
The content of the available carbohydrate was determined by the following equation
|
Carbohydrate= [100 –(Moisture + Fat + Protein + Total (g/100g sample) Ash + Crude Fibre ) ]
|
10.Determination of total lipid: 17
Total lipid was determined by slight modified method of Folch et al. (1957). Five gram of each sample was suspended in 50ml of chloroform: methanol (2:1) mixture then mixed thoroughly and let stand for 3 days. The solution was filtrated and centrifuged at 1000 RPM by a centrifuge machine. The upper layer of methanol was removed by Micro pipette and chloroform was evaporated by heating. The remaining was the crude lipid.
|
Determination = Weight of Beaker --Weight of Empty of Fat with Lipid Beaker |
11.Dry matter content:
The fresh weight (one gram) of mushroom sample was taken in a china dish using chemical balance. This was dried in the oven at 80°C for 48 h. The loss in weight obtained after drying as the final weight.
|
Dried weight
Dry matter content Initial weight |
12. Isolation of Protein from Agaricus bisporus : 14
The Agaricus bisporus (10 ml) was transferred in to the sterile centrifuge tube. The cells were pellet down by using centrifuge (10,000 rpm) for 10 minutes at -4°C. The pellets were collected, weighed and suspended in TES buffer then solublized by vortex for complete solublization. The above contents were centrifuged (10,000 rpm) for 5 minute at -4°C. The supernant was transferred to another sterile tube and the pellets were discarded. To the supernant 0.5 ml 10% TCA was added to precipitate the protein. Leave it for precipitation at 4°C for 10 minutes. Then centrifuged (10,000 rpm) for 5 minutes at -4° C. The protein was collected and 1 ml of acetone was added to remove the TCA traces. Then leave it for 10 minutes at 4° C. Then centrifuged (10,000 rpm) again for 10 minutes at -4° C. The protein was solublized by using 10 % SDS and stored at -20˚ C for further studies. The concentration of isolated protein was estimated by Lowry’s method (1951)
13. Determination of Protein content Preparation of Stock solutions:
Reagent A: - 2% Na2CO3 in 0.1N NaOH
Reagent B: - 1% Sodium Potassium Tartarate
Reagent C: - 0.5% CuSO4 in water
Reagent D: - 98ml of Reagent A + 1ml of Reagent B + 1ml of Reagent C for 100ml.
Reagent E: - Folin Reagent: distilled water (1:1) for 25ml
Procedure:
The test tube was taken and transferred 1ml of 1N NaOH solution and heated up to 100ºC. 5 µl of isolated protein was suspended in the above solution for 4 - 5 minutes. Add 5ml of reagent D (alkaline CuSO4), mixed properly and leave the mixture at room temperature for 10minutes. To the mixture 0.5ml of Folin Ciocaltue reagent was added with immediate mixing. Leave it for 30 minutes then read the absorbance at 750 nm in colorimeter. Draw standard curve (Concentration Vs Absorbance). Standard curve was obtained by using BSA for 10µg, 20µg, 30µg, 40µg, 50µg concentration and the above process was done similarly. Blank also performed for same procedure. The amount of protein was calculated from the standard curve.
14. Mineral analysis. 12
Total ash was taken for the analysis of mineral contents. Two ml of conc. HNO3 was added to the ash and heated for 2 minutes. One drop of hydrogen peroxide was added into the solution. The solution was then transferred into a volumetric flask and total volume was made 50 ml by adding deionized distilled water. This was then used to analyze the contents of calcium (Ca), potassium (K), Sodium (NA), manganese (Mn), magnesium (mg), zinc (Zn), iron (Fe), copper (Cu) and lead (Pb) by flame and graphite method with atomic absorption spectrophotometer (VARIAN SPECTRAA240, VIT, Vellore ).
RESULT:
Mushrooms are fruit bodies of macroscopic, filamentous fungi and they are made up of hyphae which forms interwoven web of tissues known as mycelium in the substrate upon which the fungus feeds. Mushrooms had long been used for medicinal and food purposes since decades. It is now increasingly recognized that correct diet, controls and modulates many functions of human body and consequently participates in the maintenance of state of good health, to reduce the risk of many diseases. Edible mushrooms are a popular vegetable for vegetarian people since they contain higher protein content than most other vegetables and are also rich in many minerals and vitamins. Mushrooms generally possess most of the attributes of nutritious food as they contain many essential nutrients in good quantity.
However, the bodies of young mushrooms are soft and brittle and therefore contain higher moisture than fully matured ones which are often tough, almost leathery and must have probably lost some of their water content. It was observed that, mushroom samples have high moisture content and high amount of ash. The various nutrients present in Agaricus bisporus showed in Table 1. Agaricus bisporus are highly valued as a good source of carbohydrate and their contents 7.652 g/100gm of dry weight 10.89%. Fresh mushrooms contain less amount of fat so they can be given to the patints suffering from hyper lipideamia and hyper cholalesterolemia. They have less carbohydrate so that they are believed to be suitable for diabetic patients.
Figure 1. Commercially purchased Agaricus bisporus
Table 1. Various nutrients present in Agaricus bisporus
|
S. NO |
Nutrients |
Contents |
|
1 2 3 4 5 6 7 8 9 10 11 |
Moisture content Ash value Acid insoluble ash Water soluble ash Sulphated ash Fat Crude fibre Carbohydrate Lipid Dry matter content Protein |
89.83% 1.355 % 99.86% 26.40% 30% 4 gm/ 100gm 2. 5gm /100 gm 7.652 g/100gm 2.38 gm / 100 gm 10. 89 % 2. 63 g/100 gm |
Mineral elements are essential for human health. The concentration of elements has an important physiological effect on different organs and cellular mechanisms. Therefore, it is necessary to know the levels of toxic and essential elements in mushrooms before using them. Table 2 shows the macronutrients and table 3 shows the micronutrients present in Agaricus bisporus. The low sodium concentration and the presence of a great quantity of potassium suggest the utilization of mushrooms in an anti-hypertensive diet; in fact, potassium from fruit and vegetables lowers blood pressure,
Table 2. Macronutrients present in Agaricus bisporus (mg/100g)
|
S.No |
Mushroom |
K |
Mg |
Ca |
Na |
|
1
|
Agaricus Bisporus |
42.2 |
30.3 |
35.1 |
22.8 |
Table 3. Micronutrients present in Agaricus bisporus (mg/100g)
|
S.No |
Mushroom |
Zn |
Fe |
Mn |
Cu |
Pb |
|
1
|
Agaricus Bisporous |
4.19 |
9.19 |
0.74 |
3.2 |
0.007 |
DISCUSSION:
Modern pharmacological research confirms large parts of traditional knowledge regarding the medicinal effects of mushrooms due to their antifungal, antibacterial, antioxidant and antiviral properties, besides being used as functional foods.17,18 Small-scale mushroom production represents an opportunity for farmers interested in an additional enterprise and is a specialty option for farmers without much land. 19 Mushroom production can play an important role in managing farm organic wastes when agricultural and food processing by-products are used as growing media for edible fungi. Due to its high percent of dry weight and high values of nutrients and mineral concentrations, Agaricus bisporus has the greater contribution of major and trace elements in the daily necessary of humans.20 This paper sums up diverse beneficial health effects of mushrooms to humans, in the form of proteins, carbohydrates, fats, lipid, vitamins, minerals, food and drugs.
Mushrooms can be cultivated in agro wastes, black soil, paper wastes and so on. They can be cultivated in a small space without sophisticated instruments, fermenters an complicated chemicals.16 Simple guidance is enough for mushroom culture. They are suitable source of protein for under nutritioned poor people.17 Mushroom cultivation converts agrowastes in to a good quality manure to enrich the fertility of the soil. Nevertheless, there is potential for an innovator who can use an existing facility, obtain a low-cost substrate, and produce a reliable supply of a high quality product. As part of a whole-farm system, mushrooms can augment productivity at any scale. Producing a nutritious food at a profit, while using materials that would otherwise be considered “waste,” constitutes a valuable service in the self-sustaining community we might envision for the future.21
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Received on 20.04.2013 Modified on 04.05.2013
Accepted on 10.05.2013 © RJPT All right reserved
Research J. Pharm. and Tech 6(7): July 2013; Page 765-768