Difference between revisions of "Part:BBa K3279007"

Revision as of 16:28, 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.

Usage and Biology

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].

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

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 <!-- Add more about the biology of this part here -->
 ===Usage and Biology===
+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].
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 <span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K3279007 SequenceAndFeatures</partinfo>
-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].