Abstract
Carbohydrate-binding modules (CBMs) are noncatalytic regions found in several enzymes of glycoside hydrolase family 13 and are proposed to orient substrates to the catalytic site. In this study, a substantial information on the conserved aromatic residues in CBM34 regions of characterized bacterial cyclolmaltodextrinases (CDases) has been presented. Molecular modeling of CDase from Geobacillus thermopakistaniensis (CDaseGt) revealed a change in the active site geometry due to CBM34 truncation. The binding energies of full-length (CDaseGt) and CBM34 truncated (CDaseGt-ΔN) models showed opposite trends. The least preferred substrate molecule by the full-length model was the most preferred by the CBM34 truncated one. These exciting in silico findings were experimentally verified by recombinant production and characterization of the full-length and the CBM34 truncated proteins. Both the enzymes showed similar optimum pH and temperature. However, substrate specificity was in the reverse order. These experimental verifications matched the homology modeling and docking predictions.
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NR designed the study and interpreted the data. MJ contributed to the construction CBM34 truncated enzyme. IA contributed to the enzyme characterization studies. NA and MA performed the experiments for in vitro and in silico characterization, respectively. All authors have seen and approved the final manuscript and its contents, and are aware of the responsibilities connected to the authorship.
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Aroob, I., Javed, M., Ahmad, N. et al. Investigating the role of carbohydrate-binding module 34 in cyclomaltodextrinase from Geobacillus thermopakistaniensis: structural and functional analyses. 3 Biotech 12, 25 (2022). https://doi.org/10.1007/s13205-021-03089-9
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DOI: https://doi.org/10.1007/s13205-021-03089-9