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| Bioactivities of Loach (Misgurnus anguillicaudatus) Skin Mucus Protein (Purification and characterization).
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A.L.M Rifky,, M.W Easther, D.M Hunupolagama, N. Suthamathy, A.M.M Irfeey
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Abstract:
Loach (Misgurnus anguillicaudatus), is a freshwater fish found in Asia, it is having a skin which is covered by a mucous layer that has a mucus structure formed by Mucin-type glycoproteins. Therefore, it is used as a traditional medicinal food to cure some human diseases. Due to its diverse biological activities, the products of fish compete to become the research demand of scholars from various countries. Therefore, the experiment was conducted to identify the availability of proteins in loach skin which is a by-product of loach processing. Three kg of live loaches with approximately the same size was placed in a basin with 300 ml of distilled water at room temperature for 10 minutes. It was allowed to produce plenty of mucus on loach skin, loaches were removed. The extracted mucus was centrifuged at a 4,200 × g for 15 minutes at 4°C. The supernatant solution was freeze-dried to obtain fine mucus powder. The powder was stored at -20°C until use. The mucus extract was purified by ion exchange chromatography and gel filtration chromatography techniques. Mucus powder was dissolved in 20 mL of 50 mMol/L Tris-HCl (pH 7.8, containing 5 mMol/L of EDTA). Two grams of mucus powder was dissolved with distilled water (200 mL) at 4°C for 2.5 hours, and then the extracts were centrifuged at 4,200 × g for 20 minutes (4°C). Then supernatant was precipitated by ethanol (supernatant: ethanol 1:4, v/v) for 24 h at 4°C. After centrifugation at 4,200 × g for 20 min (4°C), the sediment was obtained and lyophilized as crude extract. Then, the sample was loaded in DEAE-Cellulose (DE-52) column (2.6 cm×20 cm). The elution was performed at a flow rate of 30 mL/h with a linear NaCl gradient. The result showed that there are two peaks on the elusions when observed through UV spectroscopic absorbance level at 220 nm and 280 nm. Therefore, it can be concluded that the polypeptide available at the mucus could be the factor for bioactivities such as antimicrobial properties, antitumor properties.
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