单点突变Phe41→His和Phe143→Glu对在大肠杆菌中表达的重组辣根过氧化物酶折叠和催化特性的影响。
Effect of single-point mutations Phe41-->His and Phe143-->Glu on folding and catalytic properties of recombinant horseradish peroxidase expressed in E. coli.
作者信息
Gazaryan I G, Doseeva V V, Galkin A G, Tishkov V I
机构信息
Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Russian Federation.
出版信息
FEBS Lett. 1994 Nov 14;354(3):248-50. doi: 10.1016/0014-5793(94)01125-7.
Wild-type recombinant and Phe41-->His and Phe143-->Glu mutant forms of horseradish peroxidase have been expressed in E. coli and reactivated from inclusion bodies with a yield of about 25%. The purified homogeneous preparations have been studied in the reaction of ABTS oxidation. The effect of mutations on heme entrapment and kinetics of ABTS oxidation demonstrates the essential role of the replaced residues in providing the hydrophobic crevice for the non-covalent heme binding.
野生型重组辣根过氧化物酶以及苯丙氨酸41突变为组氨酸和苯丙氨酸143突变为谷氨酸的突变体形式已在大肠杆菌中表达，并从包涵体中重新激活，产率约为25%。已对纯化的均一制剂进行了ABTS氧化反应的研究。突变对血红素包封和ABTS氧化动力学的影响表明，被取代的残基在为非共价血红素结合提供疏水裂隙方面起着至关重要的作用。
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