Continuous Clarification of Barberry Juice with Pectinase Immobilised by Oxidized Polysaccharides.

RESEARCH BACKGROUND: Barberry juice is a rich source of bioactive compounds and shows different health properties such as antioxidant and anticancer activities. Clarification, as the removal process of suspended material, is an important step in the production of fruit juice due to its significant effect on the appearance, flavour and commercialisation of juice. Pectinase is the most important enzyme applied in juice clarification that breaks down the pectin polymer structure and reduces the undesirable turbidity. Pectinase immobilisation is a way to overcome free enzyme drawbacks such as instability, high cost, the difficulty of recovery and recyclability. Also, continuous clarification process which is highly preferred in fruit juice industry is not possible without enzyme immobilisation. EXPERIMENTAL APPROACH: Pectinase enzymes were immobilised on the functionalised glass beads (glass bead with (3-aminopropyl)triethoxysilane) by glutaraldehyde, polyaldehyde derivatives of pullulan and kefiran and the barberry juice was clarified in the batch and continuous processes in a packed bed reactor (PBR). Also, the effect of clarification on the physicochemical and antioxidant properties of the barberry juice samples was evaluated. RESULTS AND CONCLUSIONS: The optimum conditions for clarification in the PBR were: flow rate 0.5 mL/min, temperature 50 °C and treatment time 63 min. Clarification led to a decrease in turbidity, pH, total soluble solid content, viscosity, total phenolic content and antioxidant activity of the juice samples. Also, this process increased the clarity, acidity, reducing sugar concentration and the lightness parameter of the barberry juice. The greatest effect of clarification on the studied properties of barberry juice was related to the pectinase immobilised by the polyaldehyde of kefiran in the continuous process and both new cross-linkers (polyaldehyde derivatives of pullulan and kefiran) immobilised the enzyme better than the common cross-linker (glutaraldehyde). NOVELTY AND SCIENTIFIC CONTRIBUTION: For the first time, barberry juice was clarified with pectinase immobilised by polyaldehyde derivatives of pullulan and kefiran and the obtained results showed that the pectinase immobilisation by these new cross-linkers was much more efficient than by the glutaraldehyde as a common cross-linker. These findings can be of use for an industrialised production of fruit juices.

Immobilization of pectinase on the glass bead using polyaldehyde kefiran as a new safe cross-linker and its effect on the activity and kinetic parameters.

For the first time, polyaldehyde kefiran was applied to immobilize the pectinase on the glass bead and the finding was compared with free enzyme. Polyaldehyde kefiran, produced by periodate oxidation of kefiran, showed an aldehyde content of 23.6 ±â€¯0.9% that was confirmed by FTIR. The results showed although the optimum temperature (50 °C) was not changed by immobilization, the optimum pH was shifted from 5.0 to 5.5. In optimal conditions, the specific activity of the free and immobilized pectinase was 2.93 ±â€¯0.13 and 1.87 ±â€¯0.24 unit/mg, respectively. Also, the immobilized pectinase had a higher thermal and pH stability than free enzyme. Km and Vmax of the immobilized pectinase were higher and lower than the free enzyme, respectively. In addition, SEM and AFM images illustrated a completely non-uniform surface after enzyme immobilization on the glass bead, which seemed to be related to the polyaldehyde kefiran strands.