HISTIDINE-DIRECTED ENGINEERING OF GLYCOSAMINOGLYCAN LYASES FOR SELECTIVE CHONDROITIN SULFATE AND ALGINATE CONVERSION
DOI:
https://doi.org/10.65327/bp.v12i3.2539Keywords:
Glycosaminoglycan lyases, Histidine-directed engineering, Chondroitin sulfate, Alginate, Substrate selectivityAbstract
Targeted engineering of glycopeptidase activity and substrate specificity by histidine is a targeted approach. In this study, wild-type and engineered GAGase variants were assessed to their differential conversions of chondroitin sulfate C and alginate since secondary enzymatic activity data. The analysis comprised time-series activity profiles, concentration-response measurements, replicated absorbance at 232 nm and substrate-selectivity indices. The chondroitin sulfates were more active, while the alginate-associated cumulative activity was noticeably lower in GAGase III-H188A and GAGase III-H188N, suggesting that residue H188 plays a significant role in the recognition or productive binding of alginate. GAGase III-H398A, in contrast, showed a decrease in chondroitin sulfate activity and an increase in alginate-associated response, resulting in an almost equal selectivity profile between the two substrates. 232 nm replicated measurements showed significant chondroitin sulfate activity in both H188A and H188N, while concentration-response analysis showed different responses of alginate to H188A and H188N. Many histidine-introduced variants were more sensitive to chondroitin sulfate than were alginates. The patterns emphasize that the residue substitutions can lead to the alteration of the discrimination of the substrate without, however, eliminating catalytic competence under similar conditions. In summary, these results show that individual histidine residues have different position-specific effects on catalytic selectivity and show that it is possible to rationally design polysaccharide lyases with selective bioconversion and controlled production of oligosaccharides.
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Copyright (c) 2026 Prof. Innocent ONYESOM, Ibrahim M Awwal, Yasir Awad Ahmed, Mohammed Rabiu Abba, Revd Dr Chukwunonso J. Nosike
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