A simple method for the preparation of silica with controlled pore size, for use as a support for the immobilization of enzymes, is described in this article. Using sodium silicate and hydrochloric acid, a microporous silica was obtained that was then submitted to a hydrothermal treatment, resulting in macroporous silica suitable for enzyme immobilization. Suitability of the macroporous silica as a support depends on the method chosen for its preparation, which will determine pore volume and the effect of hydrothermal treatment on pore size. The pore volume of the support was 0.8-0.9 cc/g and the average pore size, controlled by the hydrothermal treatment, was in the range of 16 to 75 nm. The enzyme amyloglucosidase was used for the immobilization studies.

Penicillin G acylase is the second most important enzyme used by industry in an immobilized form. Penicillin hydrolysis is its main application. This reaction is used to produce 6-aminopenicillanic acid (6-APA), an intermediate in the synthesis of semisynthetic antibiotics. This work aims to compare catalytic properties of different penicillin G acylase (PGA) derivatives obtained by multipoint immobilization of the enzyme on macroporous silica. Enzyme amino groups react with different aldehyde groups produced in the support using either glutaraldehyde or glyoxyl activation. In the former method, silica reacts with <FONT FACE="Symbol">g</FONT>-aminopropyltriethoxysilane (<FONT FACE="Symbol">g</FONT>-APTS) and glutaraldehyde; in the latter, a reaction with glycidoxypropyltrimethoxysilane (GPTMS) is followed by acid hydrolysis and oxidation using sodium periodate. This work determines the influence of degree of activation, using glutaraldehyde, on immobilization parameters. PGA was immobilized on these two different supports. Maximum enzyme load, immobilized enzyme activity (derivative activity), rate of immobilization and thermal stability were checked for both cases. For glutaraldehyde activation, the results showed that 0.5% of the <FONT FACE="Symbol">g</FONT>-APTS is sufficient for all the hydroxyl groups in the silica to react. They also showed that degree of activation only affects immobilization yield and reaction velocity and that reduction of the glutaraldehyde derivatives with sodium borohydride does not affect their thermal stability. In comparing the derivatives obtained using glyoxyl and glutaraldehyde activation, it was observed that the glyoxyl derivatives presented better immobilization parameters, with a maximum enzyme load of 264 IU/g silica and a half-life of 20 minutes at 60 °C.