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There is fast growth in enzymes applications in different fields of biotechnology, bioanalytics anв medicine diagnostics. At the same time enzymes in living cell are the result of native long evolution and therefore they very often do not fit to rrrequirements of real biotechnology processes due to low activity with non native substrates, non-optimal conditions for maximum activity, low thermal and operational stability. As result, even 15-20 ago real conditions of biotechnology process were compromise between optimal process conditions and properties of biocatalysts. In present time new constrain is realized, properties of the enzyme are improved by protein engineering to fit requirements of process. In present work we will show examples of directed protein engineering as well as creation of biocatalysts with changed structure. This work was supported by Russian Foundation for Basic Research, (projects 11-04-00920, 11-04-00959, 11-04-00962-а, 12-04-31740, 13-04-01907 and 13-04-02013).