Abstract
An exo-β-d-glucosaminidase gene (PH0511) was cloned from the hyperthermophilic archaeon, Pyrococcus horikoshii, and expressed in Escherichia coli. The purified protein showed a strong exo-β-d-glucosaminidase activity by TLC analysis. DTT (50 mM) had little effect on its homodimeric structure during SDS-PAGE. The enzyme was optimally active at 90°C (over 20 min) and pH 6. It had a half-life of 9 h at 90°C and is the most thermostable glucosaminidase described up to now. The activity was not inhibited by ethanol, 2-propanol, DMSO, PEG-400, denaturing agents SDS (5%, w/v), urea, guanidine hydrochloride (5 M) and Mg2+, Mn2+, Co2+, Ca2+, Sr2+, Ni2+ (at up to 10 mM).
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This work was supported by grants from the National Natural Science Foundation of China (30570012) and the National Basic Research Program of China (2004CB719604).
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Liu, B., Li, Z., Hong, Y. et al. Cloning, expression and characterization of a thermostable exo-β-D-glucosaminidase from the hyperthermophilic archaeon Pyrococcus horikoshii . Biotechnol Lett 28, 1655–1660 (2006). https://doi.org/10.1007/s10529-006-9137-0
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DOI: https://doi.org/10.1007/s10529-006-9137-0