Now i'm finding peroxidase which can be use in high temperature (at least 60℃). most of company selling peroxidase, but it is Horseradish peroxidase and it can not use in temperature what i want to use.......
Make your own: Either steal it from a thermophilic bug or do some directed evolution.
Dear Jaeheon Kim,
The thermal stability of protein can be changed - improved - by several methods.
For first the HRP origin is mesophilic organism, so protein don't have special structural adaptation for work in higher temperatures. How mentioned Wolfgang Schechinger, You can improve properities of HRP by modyfication of its stucture by direct evolution methods, protein rational design etc., or find some similar protein with higher termal stability, express it, and use it. (For initial research I recomend You Enzyme Database - BRENDA). However it is not so easy. Another way, can by enzyme immobilisation, which methods some times improve termal stability of enzyme. The HRP is well know enzyme and was immobilized on many methods on diffrent carriers. Howewer could You more precise describe, why You need better thermal stability form of HRP ? You schould also differ in terms, the thermal stability and thermal catalytic optima of enzymes. This two parameters are not the same and some times enzyme works well in high temperatures but on short time. Example of HRP immobilisation that propably influence on its termal stability You can find here http://yadda.icm.edu.pl/yadda/element/bwmeta1.element.baztech-article-BPS2-0054-0039
The reason i want that kind of enzyme is to apply the enzyme to real-time detection of nucleic acid amplification. but seen through your answer, It seems impossible. Thanks to your kind advise Radoslaw Drozd and Wolfgang Schechinger !
Soluble and ionieally bound peroxidases have been obtained from orange juice and the albedo. A high level of peroxidase activity waspresent in the albedo. Non-linear heat inactivation was observed for orange per- oxidases. The bound peroxidases were particularly heat stable and regenerated.
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