This study investigates the capacity of phenolic extracts from microalgae Nannochloropsis sp. and Spirulina sp. to inhibit enzymes and free radical activities, intending to find an innovative way to slow down food damage. HPLC-UV and LC-MS/MS served to determine and confirm, respectively, the phenolic acid profiles in the soluble methanolic (free phenolic) and ethanolic (conjugated phenolic) fractions, and after hydrolysis (bound phenolic fractions). Different procedures measured the antioxidant activity of the extracts to estimate the minimal concentration for the protective effect, stability and versatility of activity. The ability to inhibit the oxidative process (ABTS and DPPH), α-amylase and peroxidase activities were estimated as specific inhibition (%/(min·μg)) for better comparison between the phenolic sources. The phenolic acid mass fractions in the free phenolic extracts from Spirulina sp. and Nannochloropsis sp. were 628 and 641 μg/g, respectively. Phenolic extract from Nannochloropsis sp. showed the highest value of ABTS inhibition (1.3%/(min·μg)) and highest inhibition of peroxidase activity (0.4%/(min·μg)). The extract from Spirulina sp. was a better inhibitor of α-amylase activity (0.07%/(min·μg)). Therefore, the phenolic extracts from the edible microalgae may be applied in food industry as natural protector against endogenous and exogenous hydrolytic and oxidative processes.

In this study combined cross-linked aggregates of catalase from bovine liver and glucose-oxidase from Aspergillus niger were prepared, and the effects of the precipitant and crosslinking agents, as well as the use of bovine serum albumin (BSA) as a feeder protein, on enzyme immobilization yield and thermal stability of both enzymes, were evaluated. Combi- crosslinking of enzyme aggregates (CLEAs) prepared using dimethoxyethane as precipitant, 25 mM glutaraldehyde and BSA/enzymes mass ratio of 5.45 (w/w), exhibited the highest enzyme activities and stabilities at 40 °C, pH 6.0, and 250 rpm for 5 h. The stability of both immobilized enzymes was fairly similar, eliminating one of the problems of enzyme coimmobilization. Combi-CLEAs were used in gluconic acid (GA) production in a bubble column reactor operated at 40 °C, pH 6.0 and 10 vvm of aeration, using 26 g L⁻¹ glucose as the substrate. Results showed conversion of around 96% and a reaction course very similar to the same process using free enzymes. The operational half-life was 34 h, determined from kinetic profiles and the first order inactivation model. Combi-CLEAs of glucose-oxidase and catalase were shown to be a robust biocatalyst for applications in the production of gluconic acid from glucose.