Difference between revisions of "Part:BBa K3279007"
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3279007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3279007 SequenceAndFeatures</partinfo>
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===Characterization from iGEM19_CAU_China===
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We linked β-glucosidase coding sequence into a pET30a(+) backbone, then transformed this plasmid into BL21. After the overnight (about 15h) induction under the condition of 16℃ 0.08 mM（see Fig.1[1]）, we smashed the recombinant E.coli strain with the ultrasonication to get crude enzyme solution, then we measured the enzyme activity by the method of CMC-Na(sodium carboxymethyl cellulose) assay. Considering the relatively weak cellulose degradation capacity of β-glucosidase, we mixed the crude enzyme solution with endoglucanase. The result is shown on Fig.2[1].
  
  

Revision as of 16:11, 15 October 2019


β-Glucosidase from Streptomyces coelicolor

β-Glucosidase obtained from various sources has been used in many areas, such as the enzymatic saccharification of cellulosic materials, the liberation of flavor compounds in fruit juices and wines, and so on. In cellulose hydrolysis, β-glucosidase is used to degrade cello-oligosaccharides to glucose[1]. We used this part to accomplish the cellulose degradation.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 287
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 427
    Illegal NgoMIV site found at 1103
    Illegal AgeI site found at 184
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 687
    Illegal BsaI.rc site found at 946

Characterization from iGEM19_CAU_China

We linked β-glucosidase coding sequence into a pET30a(+) backbone, then transformed this plasmid into BL21. After the overnight (about 15h) induction under the condition of 16℃ 0.08 mM（see Fig.1[1]）, we smashed the recombinant E.coli strain with the ultrasonication to get crude enzyme solution, then we measured the enzyme activity by the method of CMC-Na(sodium carboxymethyl cellulose) assay. Considering the relatively weak cellulose degradation capacity of β-glucosidase, we mixed the crude enzyme solution with endoglucanase. The result is shown on Fig.2[1].