Difference between revisions of "Part:BBa K633000"
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<partinfo>BBa_K633000 short</partinfo> | <partinfo>BBa_K633000 short</partinfo> | ||
+ | C. thermocellum endoglucanase CelD was chosen as a representative member for detailed structural and functional | ||
+ | studies of family E cellulases. Family E includes, beside C. thermocellum CelD, a number of cellulases of bacterial, fungal, and plant origin. | ||
− | + | The corresponding gene, celD, has been overexpressed in Escherichia coli, and the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies. | |
+ | This specific cellulase sequence is the result of an investigation regarding activity, several mutations directed at the carboxilic residues (aspartate, glutamate), which are involved in the catalytic mechanism of celD, finally obtaining a celD with a 224% specific activity caused by the mutation Asp-523 to Ala on the peptide sequence. | ||
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+ | Chauvaux, S., Beguin, P., & Aubert, J. (1992). Site-directed mutagenesis of essential carboxylic residues in clostridium thermocellum endoglucanase celd*. The Journal of Biological Chemistry, 267(5), 4472-4478. | ||
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===Usage and Biology=== | ===Usage and Biology=== |
Revision as of 04:53, 17 October 2011
CelD Mutated Cellulase C. thermocellum endoglucanase CelD was chosen as a representative member for detailed structural and functional studies of family E cellulases. Family E includes, beside C. thermocellum CelD, a number of cellulases of bacterial, fungal, and plant origin.
The corresponding gene, celD, has been overexpressed in Escherichia coli, and the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies.
This specific cellulase sequence is the result of an investigation regarding activity, several mutations directed at the carboxilic residues (aspartate, glutamate), which are involved in the catalytic mechanism of celD, finally obtaining a celD with a 224% specific activity caused by the mutation Asp-523 to Ala on the peptide sequence.
Chauvaux, S., Beguin, P., & Aubert, J. (1992). Site-directed mutagenesis of essential carboxylic residues in clostridium thermocellum endoglucanase celd*. The Journal of Biological Chemistry, 267(5), 4472-4478.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1831
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 644
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 508
Illegal AgeI site found at 151
Illegal AgeI site found at 567
Illegal AgeI site found at 1396 - 1000COMPATIBLE WITH RFC[1000]