Preparation and properties of alpha-galactosidase chemically attached to activated chitin.
alpha-Galactosidase (alpha-D-galactoside galactohydrolase, EC 3.2.1.22) from watermelon was covalently immobilized on chitin. The immobilized alpha-galactosidase exhibited an activity of 0.61 U per g of carrier and an activity yield of 67%. The properties of free and immobilized alpha-galactosidase were also searched and compared. The results showed that, optimum conditions for activity were not affected by immobilization. The optimum pH and temperature for free and immobilized enzyme found as pH 6.0 and 65 degress C, respectively. Compared with the free enzyme, the temperature and pH stabilities of the immobilized enzyme were similar. Both the enzymes were stable between pH 2-10 and below 50 degrees C. The Km values for free and immobilized enzyme were determined using p-nitrophenyl-alpha-D-galactopyranoside (PNPG) and raffinose as substrates. Operational stability of the immobilized enzyme was investigated by using both substrates. The operational half-life (t 1/2) was calculated as 34 h for PNPG and 28 h for raffinose. The immobilized alpha-galactosidase was also utilized in the hydrolysis of raffinose. The immobilization procedure on chitin was cheap and also easy to carry out, and the immobilized enzyme had good properties that the potential for practical application is considerable.
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Department of Biochemistry, Faculty of Science, Ege University, Bornova-Izmir, Turkey. tatar@sci.ege.edu.tr
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D002686:Chitin / Q000737:chemistry*; D029773:Citrullus / Q000201:enzymology; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007093:Imidazoles; D009598:Nitrophenylgalactosides; D019920:Photoaffinity Labels; D011887:Raffinose / Q000378:metabolism; D013439:Sulfhydryl Reagents; D013696:Temperature; D000519:alpha-Galactosidase / Q000737:chemistry* / Q000378:metabolism*
D004800:Enzymes, Immobilized; D007093:Imidazoles; D019920:Photoaffinity Labels; D013439:Sulfhydryl Reagents; D002686:Chitin; D009598:Nitrophenylgalactosides; C059200:4-nitrophenylgalactoside; C006900:N,N-carbonyldiimidazole; D000519:alpha-Galactosidase; D011887:Raffinose