Biochemical and Structural Characterisation of a Novel D-Lyxose Isomerase From the Hyperthermophilic Archaeon Thermofilum sp.
Biochemical and Structural Characterisation of a Novel D-Lyxose Isomerase From the Hyperthermophilic Archaeon Thermofilum sp.
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DOI：
10.3389/fbioe.2021.711487
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发表时间：
2021
影响因子：
5.7
通讯作者：
Littlechild JA
中科院分区：
文献类型：
--
作者：
De Rose SA;Kuprat T;Isupov MN;Reinhardt A;Schönheit P;Littlechild JA
在嗜热古生子基因组中发现了一种新的D-赖糖异构酶，属于热丝菌属。该酶已被克隆并在大肠杆菌中高效表达，并对其进行了生化特征分析。这种酶与其他这类酶的不同之处在于，它对底物D-赖糖高度专一，对甘露糖和其他已报道的赖糖异构酶的底物的活性低于2%。这是迄今报道的最具热活性和热稳定性的赖糖异构酶，其活性高于95℃，在80℃孵育60分钟后仍保持60%的活性。该赖糖异构酶在50%(v/v)的乙醇、甲醇、乙腈和DMSO存在下稳定。该酶的晶体结构在无配体形式和与两种缓慢反应的糖底物甘露糖和果糖的络合物中的分辨率为1.4-1.7毫安。这种嗜热的来聚糖异构酶通过二聚体酶的两个单体之间的二硫键和在二聚体界面上增加的疏水性来稳定。这些高底物专一性、热稳定性和耐溶剂性的综合性质使这种赖糖异构酶成为潜在的工业应用的很好候选者。
A novel D-lyxose isomerase has been identified within the genome of a hyperthermophilic archaeon belonging to the Thermofilum species. The enzyme has been cloned and over-expressed in Escherichia coli and biochemically characterised. This enzyme differs from other enzymes of this class in that it is highly specific for the substrate D-lyxose, showing less than 2% activity towards mannose and other substrates reported for lyxose isomerases. This is the most thermoactive and thermostable lyxose isomerase reported to date, showing activity above 95°C and retaining 60% of its activity after 60 min incubation at 80°C. This lyxose isomerase is stable in the presence of 50% (v/v) of solvents ethanol, methanol, acetonitrile and DMSO. The crystal structure of the enzyme has been resolved to 1.4–1.7 A. resolution in the ligand-free form and in complexes with both of the slowly reacting sugar substrates mannose and fructose. This thermophilic lyxose isomerase is stabilised by a disulfide bond between the two monomers of the dimeric enzyme and increased hydrophobicity at the dimer interface. These overall properties of high substrate specificity, thermostability and solvent tolerance make this lyxose isomerase enzyme a good candidate for potential industrial applications.
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影响因子：
5.7
作者：
Ferrandi EE;Sayer C;De Rose SA;Guazzelli E;Marchesi C;Saneei V;Isupov MN;Littlechild JA;Monti D
通讯作者：
Monti D
影响因子：
5.6
作者：
Guy, JE;Isupov, MN;Littlechild, JA
通讯作者：
Littlechild, JA
DOI：
10.1107/s0907444909042073
发表时间：
2010-01
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Chen VB;Arendall WB 3rd;Headd JJ;Keedy DA;Immormino RM;Kapral GJ;Murray LW;Richardson JS;Richardson DC
通讯作者：
Richardson DC
影响因子：
3.9
作者：
Gane, PJ;Dunwell, JM;Warwicker, J
通讯作者：
Warwicker, J
影响因子：
5.4
作者：
Hansen, T;Schlichting, B;Schönheit, P
通讯作者：
Schönheit, P