j-Carrageenan hydrogel crosslinked with protonated polyethyleneimine (PEI 1 ) and glutaraldehyde (GA) was prepared and evaluated as a novel biocatalytic support for covalent immobilization of penicillin G acylase (PGA). The method of modification of the carrageenan biopolymer is clearly illustrated using a schematic diagram and was verified by FTIR, elemental analysis, DSC, and INS-TRON using the compression mode. Results showed that the gels' mechanical strength was greatly enhanced from 3.9 kg/cm 2 to 16.8 kg/cm 2 with an outstanding improvement in the gels thermal stability. It was proven that, the control gels were completely dissolved at 358C, whereas the modified gels remained intact at 908C. The DSC thermogram revealed a shift in the endothermic band of water from 62 to 938C showing more gel-crosslinking. FTIR revealed the presence of the new functionality, aldehydic carbonyl group, at 1710 cm 21 for covalent PGA immobilization. PGA was successfully immobilized as a model industrial enzyme retaining 71% of its activity. The enzyme loading increased from 2.2 U/g (control gel) to 10 U/g using the covalent technique. The operational stability showed no loss of activity after 20 cycles. The present support could be a good candidate for the immobilization of industrial enzymes rich in amino groups, especially the thermophilic ones.

Immobilized molecules using biomaterials and nanobiotechnology is a very interesting

Grafted alginate beads were prepared using the Encapsulator by two methods, the one-step and the two-step. The methods of grafting were characterized by thermal gravimetric analysis and infrared (IR). The glass transition (Tg) of both grafted gel beads showed gradual thermal improvement over the control gel. However, the one-step method showed higher Tg (231 °C) compared to the two-step method (220 °C). Both methods were also evaluated for immobilization of an important industrial enzyme, inulinase, to produce fructose, which is good for diet regimens and suitable for diabetics. The one-step method showed an enzyme loading capacity (ELC) of 530 U/g gel beads compared to 336 U/g gel beads for the two-step method. Accordingly, the one-step method has been chosen for further optimization. The ELC has been optimized to reach 1627 U/g gel using our locally prepared crude enzyme compared to 10.9 U/g by another author using purified inulinase. The immobilization process improved as did the enzyme’s thermal stability, from 50 to 60 °C, which is the most suitable temperature used in food industries to prevent microbial contamination. The enzyme’s thermal stability test at 60 °C and for an incubation time of 2 h, revealed a drastic decrease of the free enzyme activity to 21%, compared to 89% retention of activity for the immobilized enzyme. The immobilization process improved as well the enzyme’s shelf stability, where the free enzyme lost all of its activity at room temperature after 28 days, the immobilized enzyme retained over 77% of its initial activity. These results are encouraging to produce high fructose syrup on the industrial scale as the carrier is efficient and the method is simple and economic.

Lactose has been hydrolyzed using covalently immobilized beta-galactosidase on thermally stable carrageenan coated with chitosan (hydrogel). The hydrogel's mode of interaction was proven by Fourier transform infrared spectroscopy, differential scanning calorimetry (DSC), and Schiff's base formation. The DSC thermogram proved the formation of a strong polyelectrolyte complex between carrageenan and chitosan followed by glutaraldehyde as they formed one single peak. The modification of carrageenan improved the gel's thermal stability in solutions from 35 degrees C to 95 degrees C. The hydrogel has been proven to be efficient for beta-galactosidase immobilization where 11 U/g wet gel was immobilized with 50% enzyme loading capacity. Activity and stability of free and immobilized beta-galactosidase towards pH and temperature showed marked shifts in their optimum pH from 4.5-5 to 5-5.5 and temperature from 50 degrees C to 45-55 degrees C after immobilization, which reveals higher catalytic activity and reasonable stability at wider pHs and temperatures. The apparent K(m) of the immobilized enzyme increased from 13.2 to 125 mM, whereas the V(max) increased from 3.2 to 6.6 micromol/min compared to the free enzyme, respectively. The free and immobilized enzymes showed lactose conversion of 87% and 70% at 7 h, respectively. The operational stability showed 97% retention of the enzyme activity after 15 uses, which demonstrates that the covalently immobilized enzyme is unlikely to leach. The new carrier could be suitable for immobilization of other industrial enzymes.

Inulinase has been extracted from Penicillium chrysogenum P36 and immobilized on a novel matrix of grafted biopolymer. The crude enzyme has been characterized in terms of specific activity, optimum temperature, and temperature stabilities. A novel matrix of alginate modified with polyimines and cross-linked with glutaraldehyde was prepared in beads shape using the Encapsulator to covalently immobilize crude inulinase. The modified beads were characterized using the FTIR and the DSC techniques. The FTIR showed the presence of the aldehydic’s carbonyl group at 1670 cm−1, which differs from that of the carboxylic group at 1620 cm−1. The DSC revealed a significant improvement of the gel’s thermal stability from 200 to 240 °C. The immobilization process improved the enzyme’s optimum temperature from 50 to 55 °C as well as the enzyme’s thermal stability for 2 h at 60 °C with 78% retention of activity as compared to only 7% for the free enzyme. The enzyme’s optimum pH slightly shifted from pH 4.8 for the free enzyme to pH 5 for the immobilized enzyme. However, at pH 5.2−5.5, the enzyme activity improved from 39% for the free enzyme to 75% for the immobilized enzyme. The novel matrix successfully immobilized the inulinase covalently with an enzyme loading capacity of 461 U/g gel. The reusability test proved the durability of the grafted alginate for 20 cycles with retention of 95% of the immobilized enzyme activity, whereas the untreated alginate gel completely dissolved by the eighth use. The results were promising; the grafting method is simple, and immobilization efficiency and enzyme loading capacity could be further improved by optimizing the gel beads’ formulations and the conditions of immobilization for the industrial applications.