从嗜热地芽孢杆菌 T2 中克隆、表达和生化特性分析一种新型的中温耐热 GDSL 家族酯酶。
Cloning, expression and biochemical characterization of a novel, moderately thermostable GDSL family esterase from Geobacillus thermodenitrificans T2.
机构信息
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.
出版信息
J Biosci Bioeng. 2013 Feb;115(2):133-7. doi: 10.1016/j.jbiosc.2012.08.016. Epub 2012 Sep 23.
A thermostable GDSL family esterase-encoding gene, EstL5, was directly obtained from the genomic DNA of Geobacillus thermodenitrificans T2. Recombinant hexahistidine-tagged EstL5 was overexpressed, purified, and its biochemical properties were partially characterized. EstL5 was observed to be active within the temperature range of 0-80°C, having maximal activity at 60°C. Unlike most other thermostable enzymes, EstL5 displayed 24% of its highest activity at 0°C. EstL5 exhibited a high level of activity within a pH range of 6.0-11.0, showing the highest activity at pH 8.0. EstL5 also retained 100% of its activity after a 12-h incubation at 55°C. Furthermore, this enzyme was observed to be strongly inhibited by 10% (w/v) SDS and 0.1 mM PMSF.
直接从地芽孢杆菌 T2 的基因组 DNA 中获得了一种热稳定 GDSL 家族酯酶编码基因 EstL5。表达、纯化了重组六组氨酸标记的 EstL5，并对其生化性质进行了部分表征。观察到 EstL5在 0-80°C 的温度范围内具有活性，在 60°C 时具有最大活性。与大多数其他耐热酶不同，EstL5 在 0°C 时表现出其最高活性的 24%。EstL5 在 pH 值为 6.0-11.0 的范围内表现出高活性，在 pH 值为 8.0 时表现出最高活性。EstL5 在 55°C 孵育 12 小时后仍保持 100%的活性。此外，该酶被 10%（w/v）SDS 和 0.1 mM PMSF 强烈抑制。
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