Abstract
Tannin acyl hydrolase produced extracellularly by the fungal strain Penicillium notatum NCIM 923 in mixed solid state fermentation of wheat bran and marigold flower in the ratio 4 : 1 was purified from the cell-free extract broth by ammonium sulphate fractionation followed by diethylaminoethyl-cellulose column chromatography. Tannase was purified by 19.89-fold with yield of 11.77%. The specific activity of crude tannase was found to be 1.31 U/mg protein while that of purified tannase was 22.48 U/mg protein. SDS-PAGE analysis indicated that the enzyme is dimeric with one major band of molecular mass 97 kDa and a very light band of molecular mass 43 kDa. Temperature of 35 to 40°C and pH 5 were optimum for tannase activity. The enzyme retained more than 60% of its stability at 60°C and 40% stability at pH 3 and 8, respectively. K m was found to be 0.33 × 10−2 M and V max = 40 U/mg. Since the enzyme is active over a wide range of pH and temperature, it could find potential use in the food processing industry.
Highlights
Tannins are phenolic compounds, which can be grouped as hydrolysable and nonhydrolyzable tannins
The present study describes purification and characterization of tannase produced by Penicillium notatum National Collection of Industrial Microorganisms (NCIM) 923 whose biochemical properties may render it of commercial interest
Penicillium notatum NCIM 923 was collected from National Collection of Industrial Microorganisms (NCIM), National Chemical Laboratory, Pune (India), and was maintained on Czapek Dox agar slants of composition—Glucose 5%, NaNO3 0.2%, KCl 0.05%, MgSO4⋅7H2O 0.05%, FeSO4⋅7H2O 0.001%, KH2PO4 0.1%, Agar 3% pH 5
Summary
Tannins are phenolic compounds, which can be grouped as hydrolysable and nonhydrolyzable tannins. Tannin acyl hydrolase (E.C.3.1.20), commonly named tannase, only hydrolyses the hydrolysable tannins and catalyses the hydrolysis of ester and depside bonds in tannic acid releasing glucose and gallic acid [1]. It has extensive applications in the food, feed, pharmaceutical, beverage, and chemical industries and is extensively used in wine, beer, and coffee-flavored soft drinks or as an additive in food detannification [2]. The purification and characterization of tannase had been attempted owing to its applications in various food, feed, leather, and pharmaceutical industries Techniques such as affinity chromatography, ultrafiltration, high performance liquid chromatography, and electrophoresis. The present study describes purification and characterization of tannase produced by Penicillium notatum NCIM 923 whose biochemical properties may render it of commercial interest
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