Enzyme immobilization in MANAE supports under different conditions
First, we tried to immobilize the enzyme at pHs 5, 7 and 9 in MANAE agarose using 5 mM of buffer. To control the pH, the support needs to be washed with a large volume of the immobilization buffer, otherwise the buffering properties of the support make the immobilization pH very similar between runs, because the concentration of amino groups in the support may be very high compared to the concentration of the buffer [3]. After this precaution, Fig. 1 shows that the enzyme may be immobilized on MANAE agarose at the 3 pH values, in only a few minutes, and maintaining over 80% of activity at all studied pH values.