Purification of pineapple bromelain by IMAC chromatography using chlorophyll-activated macroporous matrices.

This study investigated the purification of bromelain obtained from pineapple fruit using a new adsorbent for immobilized metal ion affinity chromatography (IMAC), with chlorophyll obtained from plant leaves as a chelating agent. The purification of bromelain was evaluated in batches from the crude extract of pineapple pulp (EXT), and the extract precipitated with 50 % ammonium sulfate (EXT.PR), the imidazole buffer (200 mM, pH 7.2) being analyzed and sodium acetate buffer, pH 5.0 + 1.0 NaCl as elution solutions. All methods tested could separate forms of bromelain with molecular weights between ±21 to 25 kDa. Although the technique using EXT.PR stood out in terms of purity, presenting a purification factor of around 3.09 ± 0.31 for elution with imidazole and 4.23 ± 0.12 for acetate buffer solution. In contrast, the EXT methods obtained values between 2.44 ± 0.23 and 3.21 ± 0.74 for elution with imidazole and acetate buffer, respectively, for purification from EXT.PR has lower yield values (around 5 %) than EXT (around 15 %). The number of steps tends to reduce yield and increase process costs, so the purification process in a monolithic bed coupled to the chromatographic system using the crude extract was evaluated. The final product obtained had a purification factor of 6, with a specific enzymatic activity of 59.61 ± 0.00 U·mg-1 and a yield of around 39 %, with only one band observed in the SDS-PAGE electrophoresis analysis, indicating that the matrix produced can separate specific proteins from the total fraction in the raw material. The IMAC matrix immobilized with chlorophyll proved promising and viable for application in protease purification processes.

Characterization of starch-based bioplastics from jackfruit seed plasticized with glycerol.

Biodegradable films based on starches from different botanical sources exhibited physicochemical and functional properties which were related with the starch characteristics. However, had inadequate mechanical properties and were hard and brittle. In this research, jackfruit seed starch plasticized with glycerol were developed and characterized. The starch and glycerol concentrations ranged from 2 to 6% w/w and 20 to 60 g/100 g starch, respectively. Bioplastics were obtained by the casting method and characterized in terms of color, mechanical properties, solubility, water vapor permeability (WVP), morphology and free energy of the hydrophobic interaction. Electronic micrographics showed the presence of some intact starch granules. The bioplastics were hydrophilic and those of 6% starch and 40% glycerol were the most hydrophilic ([Formula: see text] = 41.35 mJ m-1). The solubility of the films presented a direct relationship with the starch concentration ranging from 16.42 to 23.26%. Increased opacity and color difference were observed with increasing starch concentration. The WVP ranged from 1.374 × 10-3 to 3.07 × 10-4 g m/day m2 which was positively related with the concentration of starch and glycerol. Tensile strength, percent elongation and Young's Modulus indicated that the jackfruit starch and glycerol provided a film with good mechanical properties. The results replaced that jackfruit starch can be used to develop films, with low opacity, moderate WVP and relatively high mechanical stability, by using glycerol in the gelatinized starch dispersions.