Subunit analyses of a novel thermostable glucose dehydrogenase showing different temperature properties according to its quaternary structure

被引:31
作者
Yamazaki, T [1 ]
Tsugawa, W [1 ]
Sode, K [1 ]
机构
[1] Tokyo Univ Agr & Technol, Fac Technol, Dept Biotechnol, Tokyo 1848588, Japan
关键词
glucose dehydrogenase; thermal stability; catalytic subunit; electron transfer subunit; quaternary structures;
D O I
10.1385/ABAB:77:1-3:325
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We previously reported a novel glucose dehydrogenase (GDH) showing two peaks in the optimum temperature for the reaction at around 45 degrees C and at around 75 degrees C. Each peak derived from hetero-oligomeric enzyme, constructed from two distinct peptides with an alpha-subunit (MWs 67,000) and beta-subunit (MWs 43,000), and a single peptide enzyme containing an alpha-subunit alone. The function of the two subunits in the thermostable co-factor binding GDH was investigated. The results of spectroscopic analyses indicated that the alpha-subunit contained an unknown co-factor showing specific fluorescence spectra like pyrroloquinoline quinone (PQQ), and the beta-subunit was cytochrome c. Moreover, the results of a urea denaturation and reconstitution experiment suggested that the dissociation of the hetero-oligomeric complex to a single peptide was reversible. The kinetic parameter analyses for glucose and the electron mediator also suggested that the beta-subunit was responsible for electron transfer from the catalytic tenter of the alpha-subunit to the electron mediator.
引用
收藏
页码:325 / 335
页数:11
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