Applications of gellan natural polymer microspheres in recombinant catechol-O-methyltransferase direct capture from a Komagataella pastoris lysate

The present work shows the application of nickel- and magnesium-crosslinked gellan microspheres in ionic and affinity capture strategies to directly extract hSCOMT from the complex Komagataella pastoris lysate through a simple batch method. Both formulations present similar morphology, but nickel-crosslinked microspheres present higher crosslinker content and smaller diameters. Four different capture strategies were established, by manipulating the ionic strength, pH. temperature and competing agents' presence. The most promising results for hSCOMT capture and clarification were obtained employing an ionic strategy with nickel-crosslinked microspheres and an affinity strategy with magnesium-crosslinked microspheres at 4 degrees C. The bioactivity results (200%) and purification degree (70%) of hSCOMT captured by the ionic strategy were more satisfactory probably due to the soft ionic conditions used (100 mM NaCl). For the first time, the gellan polysaccharide versatility was demonstrated in the microsphe re application for the direct capture of hSCOMT from a complex lysate, simplifying isolation biotechnological procedures. (C) 2021 Elsevier B.V. All rights reserved.

Automated summary

This study demonstrates the direct extraction of hSCOMT from a complex lysate using different capture strategies, with the ionic strategy showing the most promising results in terms of bioactivity and purification degree. The versatility of gellan polysaccharide in microsphere application simplifies the biotechnological procedures for isolating hSCOMT.