Mutation Analysis of a Thermostable Exo-ß-D-Glucosaminidase from the Hyperthermophilic Archaeon Pyrococcus Horikoshii and Comparison with Two Mesophilic Exo-ß-D-Glucosaminidases

Abstract:

The mechanism of thermostabilization of the thermostable exo-ß-D-glucosaminidase (Bglph) from Pyrococcus horikoshii was investigated. Conserved domains analysis and C-terminal truncated mutation showed that the C-terminal region was indispensable for the enzymes thermostability. Two site directed mutagenesis proteins (C103A and C103AandC550A) were as active as the native enzyme and showed no structural difference with the native enzyme. By comparison with two mesophilic exo-ß-D-glucosaminidases (CsxA and CsxT) from actinomycetes Amycolatopsis orientalis and fungi Trichoderma reesei respectively, it was shown that the amino acids composition of Bglph was consistent with its high thermostability, hydrophobic interactions, homodimerization and ion pairs could play key roles. These results presented the overall properties of the thermostability of Bglph from diverse aspects.

• Periodical: Advanced Materials Research (Volume 787)
• Pages: 172-176
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.787.172
• Online since: September 2013
• Authors: Hui Hui Shi, Yan Yan Gao, Tian Ming Liu, Bo Liu
• Keywords: Bglph, C-Terminal Truncated Mutation, Homodimer, Hydrophobic Interaction, Ion Pairs, Thermostability

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