Novel Biocatalysts by Identification and Design |
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Authors: | Thorsten Eggert Christian Leggewie Michael Puls Wolfgang Streit Gertie van Pouderoyen Bauke W. Dijkstra Karl-erich Jaeger |
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Abstract: | Enzymes produced from bacteria and eukaryotic organisms are presently being used for a large variety of different biotechnological applications. The rapidly increasing demand for enzymes which are active towards novel and often non-natural substrates has triggered the development of novel molecular biological methods of enzyme isolation and design. The metagenome approach is a cultivation-independent method which allows the direct cloning and expression of environmental DNA thereby providing access to a wealth of so-far unknown biocatalysts. Additionally, newly identified or existing biocatalysts can be further optimized by different methods of directed evolution. Here, the principle of the metagenome approach is outlined and a strategy is presented for the optimization of a bacterial lipase using a combination of rational design and directed evolution. |
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Keywords: | Bacillus subtilis lipase biodiversity directed evolution metagenome |
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