Kulkarni Dhananjay S, Kapanoor Shankar S, Girigouda Kotiguda, Kote Naganagouda V, Mulimani Veerappa H
Department of Biochemistry, Gulbarga University, Gulbarga-585 106, Karnataka, India.
Biotechnol Appl Biochem. 2006 Sep;45(Pt 2):51-7. doi: 10.1042/BA20060027.
Alpha-galactosidase from Aspergillus oryzae was immobilized on chitosan beads using glutaraldehyde as a cross-linking agent. The general properties of free and immobilized enzymes were determined. The optimum pH for the free and immobilized enzymes was 4.8 and 4.6 respectively. The optimum temperature for the free enzyme was 50 degrees C, whereas that of immobilized enzyme was increased to 56 degrees C. Kinetic parameters were determined with synthetic substrate (p-nitrophenyl alpha-D-galactopyranoside) and raffinose. Immobilized enzyme showed a higher Km and a lower Vmax than the free enzyme. The immobilized enzymes were used in batch, repeated and continuous mode. A level of 92% hydrolysis was observed at a flow rate of 60 ml/h. The immobilized enzyme was used repeatedly ten times without any change in the performance of the immobilized enzyme in fluidized-bed reactor. The results obtained are of considerable interest for industrial purposes.
使用戊二醛作为交联剂,将米曲霉的α-半乳糖苷酶固定在壳聚糖珠上。测定了游离酶和固定化酶的一般性质。游离酶和固定化酶的最适pH分别为4.8和4.6。游离酶的最适温度为50℃,而固定化酶的最适温度提高到56℃。用合成底物(对硝基苯基α-D-吡喃半乳糖苷)和棉子糖测定动力学参数。固定化酶比游离酶表现出更高的Km值和更低的Vmax值。固定化酶以分批、重复和连续模式使用。在流速为60 ml/h时观察到92%的水解水平。固定化酶在流化床反应器中重复使用十次,固定化酶的性能没有任何变化。所得结果对于工业用途具有相当大的意义。