State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, People's Republic of China.
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。
