Behaviour of horseradish peroxidase in AOT reversed micelles

被引:13
|
作者
Azevedo, AM [1 ]
Fonseca, LP [1 ]
Graham, D [1 ]
Cabral, JMS [1 ]
Prazeres, DMF [1 ]
机构
[1] Univ Tecn Lisboa, Ctr Engn Biol & Quim, Inst Super Tecn, P-1049001 Lisbon, Portugal
关键词
horseradish peroxidase; reversed micelles; thermostability;
D O I
10.3109/10242420109105265
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
The activity and stability of horseradish peroxidase (HRP) solubilised in AOT reversed micelles in isooctane and decalin was studied using guaiacol (2-methoxyphenol) as the electron donor. The activity of the enzyme in both reversed micellar systems increases with the water content until reaching a maximum value that remains fairly constant for water contents higher than 3.05% (v/v) in isooctane and 2.20% in decalin. The effect of pH on the activity profile was studied in the system AOT/isooctane. The enzyme is fully active at pH 7 and 8 for water contents higher than 3.05% (v/v) but it was completely deactivated at pH 9. The effect of surfactant concentration on HRP activity was also investigated. At low water contents a strong dependence was observed, whilst no further activity increase was observed for water content values higher than 2.7% (v/v). The stability of HRP was found to be strongly dependent on the water content of the system with higher levels of stability obtained for higher values of water content. HRP stability is also affected by the presence of substrates. Whilst the stability increases markedly when the enzyme is incubated with guaiacol, it does not appear to be so strongly affected by the presence of hydrogen peroxide, at the concentrations studied.
引用
收藏
页码:213 / 233
页数:21
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