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<partinfo>BBa_K633000 short</partinfo>
 
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C. thermocellum endoglucanase CelD was chosen as a representative member for detailed structural and functional
 
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studies of family E cellulases. Family E includes, beside C. thermocellum CelD, a number of cellulases of bacterial, fungal, and plant origin.
 
  
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The corresponding gene, celD, has been overexpressed in Escherichia coli, and the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies.
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''Clostridium thermocellum'' endoglucanase CelD is a representative enzyme for detailed structural and functional studies of the family E cellulases.
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Beside ''C. thermocellum’s'' CelD, E family cellulases include a number of cellulases of bacterial, fungal, and plant origin.
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CelD, has been previously overexpressed in ''Escherichia coli'', the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies. (Chauvaux, Beguin, Aubert, 1992)
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The CelD sequence used has an aminoacid substitution Asp-523 ---> Ala. That mutation increases the specific activity of the enzyme by 224% (Chauvaux, Beguin, Aubert, 1992)
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<center>[[Image:Celd_chart.png]]__NOTOC__</center>
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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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References
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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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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===

Latest revision as of 01:05, 20 October 2011

CelD Mutated Cellulase



Clostridium thermocellum endoglucanase CelD is a representative enzyme for detailed structural and functional studies of the family E cellulases.
Beside C. thermocellum’s CelD, E family cellulases include a number of cellulases of bacterial, fungal, and plant origin.
CelD, has been previously overexpressed in Escherichia coli, the enzyme is easily purified in large amounts from cytoplasmic inclusion bodies. (Chauvaux, Beguin, Aubert, 1992)
The CelD sequence used has an aminoacid substitution Asp-523 ---> Ala. That mutation increases the specific activity of the enzyme by 224% (Chauvaux, Beguin, Aubert, 1992)

Celd chart.png






References
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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1831
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 644
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]