Thermal adaptation of dihydrofolate reductase from the moderate thermophile Geobacillus stearothermophilus.

Thermal adaptation of dihydrofolate reductase from the moderate thermophile Geobacillus stearothermophilus.
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DOI:
10.1021/bi500238q
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发表时间:
2014-05-06
期刊:
影响因子:
2.9
通讯作者:
Allemann RK
Allemann RK
中科院分区:
生物学3区
文献类型:
--
作者:
Guo J;Luk LY;Loveridge EJ;Allemann RK

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嗜热脂肪芽孢杆菌二氢叶酸还原酶的热熔化温度比嗜冷菌的同源物高∼30℃。有人建议在BsDHFR的环区增加脯氨酸残基,以提高BsDHFR的热稳定性,但定点突变研究表明,这些脯氨酸残基对热稳定性的贡献很小。相反,BsDHFR的高热稳定性部分是由于去除了可接触水的不耐热残留物,如谷氨酰胺和蛋氨酸,这些残留物在高温下容易水解或氧化。BsDHFR的额外热稳定性可以通过配体结合或在盐或助溶剂(如甘油和蔗糖)的存在下获得。所有这些增量因素的总和使BsDHFR能够在G.stearthermophilus的自然栖息地高效运作,该栖息地的特点是温度可达75摄氏度。
The thermal melting temperature of dihydrofolate reductase from Geobacillus stearothermophilus (BsDHFR) is ∼30 °C higher than that of its homologue from the psychrophile Moritella profunda. Additional proline residues in the loop regions of BsDHFR have been proposed to enhance the thermostability of BsDHFR, but site-directed mutagenesis studies reveal that these proline residues contribute only minimally. Instead, the high thermal stability of BsDHFR is partly due to removal of water-accessible thermolabile residues such as glutamine and methionine, which are prone to hydrolysis or oxidation at high temperatures. The extra thermostability of BsDHFR can be obtained by ligand binding, or in the presence of salts or cosolvents such as glycerol and sucrose. The sum of all these incremental factors allows BsDHFR to function efficiently in the natural habitat of G. stearothermophilus, which is characterized by temperatures that can reach 75 °C.
DOI: 10.1039/c39790000922
发表时间: 1979-01-01
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