Exceptional thermal stability and organic solvent tolerance of an esterase expressed from a thermophilic host |
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Authors: | Yuxia?Mei Nan?Peng Shumiao?Zhao Yongmei?Hu Huacai?Wang Email author" target="_blank">Yunxiang?LiangEmail author Email author" target="_blank">Qunxin?SheEmail author |
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Institution: | 1.State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology,Huazhong Agricultural University,Wuhan,People’s Republic of China;2.Archaeal Centre, Department of Biology,University of Copenhagen,Copenhagen N,Denmark |
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Abstract: | A protein expression system recently developed for the thermophilic crenarchaeon Sulfolobus islandicus was employed to produce recombinant protein for EstA, a thermophilic esterase encoded in the same organism. Large amounts
of protein were readily obtained by an affinity protein purification, giving SisEstA. Upon Escherichia coli expression, only the thioredoxin-tagged EstA recombinant protein was soluble. The fusion protein was then purified, and removing
the protein tag yielded EcSisEstA. Both forms of the thermophilic EstA enzyme were characterized. We found that SisEstA formed
dimer exclusively in solution, whereas EcSisEstA appeared solely as monomer. The former exhibited a stronger resistance to
organic solvents than the latter in general, having a much higher temperature optimum (90°C vs. 65°C). More strikingly, SisEstA
exhibited a half-life that was more than 32-fold longer than that of EcSisEstA at 90°C. This indicated that thermophilic enzymes
yielded from homologous expression should be better biocatalysts than those obtained from mesophilic expression. |
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