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Article Abstract
Two multimodular endoglucanases in glycoside hydrolase family 5, Cel5 and Cel5, share 73% identity and exhibit similar modular structures: family 1 carbohydrate-binding module (CBM1); catalytic domain; CBMX2; module of unknown function. However, they differed in their biochemical properties and catalytic performance. Cel5 showed optimal activity at pH 4.0 and 70 °C, maintaining stability at 70 °C (>80% activity). Conversely, Cel5 is optimal at pH 3.0 and 50 °C (>50% activity at 50 °C). Cel5 excels in degrading CMC-Na (256 U/mg vs 53 U/mg of Cel5). Five domain-truncated (TM1-TM5) and four domain-replaced (RM1-RM4) mutants of Cel5 with the counterparts of Cel5 were constructed, and their enzymatic properties were compared with those of the wild type. Only RM1, with Cel5-CBM1, displayed enhanced thermostability and activity. The hydrolysis of pretreated corn stover was reduced in most TM and RM mutants. Molecular dynamics simulations revealed interdomain interactions within the multimodular endoglucanase, potentially affecting its structural stability and complex biological catalytic processes.
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Article Synopsis
- Two multimodular endoglucanases, Cel5 and Cel5, have a 73% identity and a similar structure but differ in their biochemical performance, with Cel5 being more effective in degrading CMC-Na.
- Cel5 operates best at pH 4.0 and 70 °C, while Cel5 is optimal at pH 3.0 and 50 °C, showing that Cel5 has better thermal stability and activity under higher temperatures.
- Mutants of Cel5 were created to study their properties, revealing that only RM1, which retains a specific module, displayed improved thermostability, while most modifications led to decreased hydrolysis efficiency on pretreated corn stover.
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