Difference between revisions of "Part:BBa K1360001"
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__NOTOC__
 
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<partinfo></partinfo>
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<partinfo>BBa_K1360001 short</partinfo>
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This part is the coding sequence for a thermostable alpha-L-arabinofuranosidase, which specifically releases arabinofuranoside branch of arabinoxylan from xylan.
  
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Xylanases are enzymes that degrad Xylan, a major component of wood. This is the circular counterpart of <partinfo>BBa_K1362020</partinfo>. A circular Xylanase should be resistent to exo-peptidases and could be more heat stable because of its reduced conformational space. This could be interesting for industrial applications, as they are often performed at higher temperatures and harsher environments.
 
  
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[[File:DNS_Abstract.png|center|frame||caption]]
 
 
===Usage and Biology===
 
===Usage and Biology===
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To characterize our part in respect to its thermal stability we conducted a Heatshock Assay. Firstly the Xylanases were incubated at 37 °C, 50 °C, 56 °C, 60 °C and 66 °C. After 30 minutes substrate (EnzChek® Ultra Xylanase Assay Kit) was added and the fluorescence was measured for 150 minutes in a platereader at 37 °C. The results indicate a reduction of activity at temperatures higher than 50 °C and complete loss of function for temperatures higher than 60 °C.
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The ArfB enzyme produced by this part can be used in pulp bleaching. It presents high activity under a wide range of temperature (55℃-99℃) and pH conditions (3-10.5). The optimized working condition is pH = 4.0, temperature = 71℃.
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===Design considerations===
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This part is only a coding sequence without expression regulators, and is compatible with RFC10 standards, and the codon is optimized for E.coli expression.
  
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===Lab Archives===
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[[File:DNS_Abstract.png|center|frame||We use DNS to test the enzymatic activity. DNS is a reagent detecting reductive saccharides. When mannan or xylan is degraded into mannose or xylose, DNS color get dark. When DNS color darker, the enzymatic activity higher.]]
  
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1362022 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1360001 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K1362022 parameters</partinfo>
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<partinfo>BBa_K1360001 parameters</partinfo>
 
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Revision as of 13:53, 1 November 2014

alpha-L-arabinofuranosidase ArfB

This part is the coding sequence for a thermostable alpha-L-arabinofuranosidase, which specifically releases arabinofuranoside branch of arabinoxylan from xylan.


Usage and Biology

The ArfB enzyme produced by this part can be used in pulp bleaching. It presents high activity under a wide range of temperature (55℃-99℃) and pH conditions (3-10.5). The optimized working condition is pH = 4.0, temperature = 71℃.

Design considerations

This part is only a coding sequence without expression regulators, and is compatible with RFC10 standards, and the codon is optimized for E.coli expression.

Lab Archives

We use DNS to test the enzymatic activity. DNS is a reagent detecting reductive saccharides. When mannan or xylan is degraded into mannose or xylose, DNS color get dark. When DNS color darker, the enzymatic activity higher.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 679
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1156
    Illegal BsaI.rc site found at 1282
    Illegal BsaI.rc site found at 1372