Proteinové inženýrství halogenalkandehalogenasy LinB změna reakčního mechanismu /
Haloalkane dehalogenases are microbial hydrolases catalyzing removal of halogens from aliphatic hydrocarbons. Based on structural and sequence similarities between haloalkane dehalogenase LinB and luciferase from Renilla reniformis, we propose that it could be possible to construct a mutant of LinB...
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| Typ dokumentu: | VŠ práce nebo rukopis |
| Jazyk: | Angličtina |
| Vydáno: |
2008
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| On-line přístup: | http://is.muni.cz/th/229688/prif_m/ |
| Shrnutí: | Haloalkane dehalogenases are microbial hydrolases catalyzing removal of halogens from aliphatic hydrocarbons. Based on structural and sequence similarities between haloalkane dehalogenase LinB and luciferase from Renilla reniformis, we propose that it could be possible to construct a mutant of LinB with luminescence activity or to obtain an enzyme with both luminescence and dehalogenase activities. This Diploma Thesis is focused on selection of the LinB variant suitable for directed evolution towards luminescence activity from previously prepared mutants LinB53, LinB54, LinB55 and LinB56. Based on the level of expression, solubility, folding, and activity of mutants, the variants LinB54 and LinB55 were chosen for fine-tuning by directed evolution. Sensitive high-throughput screening, which is an important prerequisite for directed evolution experiments, was developed. The screening uses the multi-well plates format and can be applied to the cell lysates. The optimization of the testing. |
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| Popis jednotky: | Vedoucí práce: Jiří Damborský |
| Fyzický popis: | 73 l. : il., obr. |