Difference between revisions of "Part:BBa K2139017"
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− | Cellulomonas fimi uses 3 endoglucanases (including CenA, accession M15823) and an exoglucanase in the degradation of cellulose into cellobiose, before using beta-glucosidase to catalyse the conversion of cellobiose to D-glucose. | + | Cellulomonas fimi uses 3 endoglucanases (including CenA, accession M15823) and an exoglucanase in the degradation of cellulose into cellobiose, before using beta-glucosidase to catalyse the conversion of cellobiose to D-glucose. This part was codon optimized for the expression in Caulobacter crescentus and synthesized by IDT. |
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+ | We improved the characterization of this part by optimizing the expression and attempted to express the enzyme on the surface layer of <i>C.crescentus</i>. We characterized the activity of the CenA insert in the <i>C.crescentus</i> s-layer. In initial activity analysis we saw no increased activity compared to wild type RsaA protein indicating that either the protein insert as we designed it does not fold correctly in the surface layer, further cloning attempts and design alterations must be made to try to improve the results. | ||
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+ | [[Image:British_Columbia.jpg|300px]]</center> | ||
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Latest revision as of 22:05, 30 October 2016
Coding Sequence of CenA
Cellulomonas fimi uses 3 endoglucanases (including CenA, accession M15823) and an exoglucanase in the degradation of cellulose into cellobiose, before using beta-glucosidase to catalyse the conversion of cellobiose to D-glucose. This part was codon optimized for the expression in Caulobacter crescentus and synthesized by IDT.
We improved the characterization of this part by optimizing the expression and attempted to express the enzyme on the surface layer of C.crescentus. We characterized the activity of the CenA insert in the C.crescentus s-layer. In initial activity analysis we saw no increased activity compared to wild type RsaA protein indicating that either the protein insert as we designed it does not fold correctly in the surface layer, further cloning attempts and design alterations must be made to try to improve the results.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 391
Illegal NgoMIV site found at 723 - 1000COMPATIBLE WITH RFC[1000]