Difference between revisions of "Part:BBa K118022"
| Line 29: | Line 29: | ||
==Contribution == | ==Contribution == | ||
| − | *'''Group:''' | + | *'''Group:''' [https://2019.igem.org/Team:CAU_China iGEM Team CAU-China 2019] |
*'''Author:''' Liang siwen, Yan xueshan | *'''Author:''' Liang siwen, Yan xueshan | ||
*'''Summary:''' Enzyme activity assay (CMC-Na as substrate) | *'''Summary:''' Enzyme activity assay (CMC-Na as substrate) | ||
===Characterization from iGEM19_CAU_China=== | ===Characterization from iGEM19_CAU_China=== | ||
| − | We linked the cex gene BBa_K118022 into a pET30a(+) backbone, then transformed this plasmid into BL21. After the overnight (about 15h) induction under the condition of 16℃ 0.08 mM, 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) assey. The result is shown on Fig.1 | + | We linked the cex gene BBa_K118022 into a pET30a(+) backbone, then transformed this plasmid into BL21. After the overnight (about 15h) induction under the condition of 16℃ 0.08 mM, 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) assey. The result is shown on Fig.1. |
===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K118022 parameters</partinfo> | <partinfo>BBa_K118022 parameters</partinfo> | ||
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Revision as of 06:21, 16 October 2019
cex coding sequence encoding Cellulomonas fimi exoglucanase
The cellulolytic bacterium Cellulomonas fimi uses an exoglucanase (from cex, accession M15824) along with 3 endoglucanases in the degradation of cellulose into cellobiose, before use B-glucosidase to catalyse the conversion of cellobiose to D-glucose.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 524
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 157
Illegal NgoMIV site found at 530
Illegal NgoMIV site found at 1032 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 577
Illegal SapI.rc site found at 660
Contribution
- Group: [http://2018.igem.org/Team:UESTC-China iGEM Team UESTC-China 2018]
- Author: Liang Zhao, Yetao Zou
- Summary: Enzyme digestion and enzyme activity assay
Characterization from iGEM18-UESTC-China
Molecular weight
This gene codes for a protein of 485 amino acids with a molecular mass of 51.2 kDa.
Enzyme digestion
We did a codon optimization of this part before using it. And we verified it by enzyme digestion.
Filter paper assay
We constructed a plasmid containing cenA gene BBa_K118023, cex gene BBa_K118022. We transformed this plasmid into BL21(DE3). We used an intracellular fraction (crude enzyme solution) obtained by ultrasonication to carry out an experiment for measuring total enzyme activities by the method of filter paper assay.The result is shown on Fig. 2[1].
References
[1]Luciano Silveira MH, Rau M, Pinto da Silva Bon E & Andreaus J. 2012. A simple and fast method for the determination of endo- and exo-cellulase activity in cellulase preparations using filter paper. Enzyme and Microbial Technology, 51: 280-285.
Contribution
- Group: iGEM Team CAU-China 2019
- Author: Liang siwen, Yan xueshan
- Summary: Enzyme activity assay (CMC-Na as substrate)
Characterization from iGEM19_CAU_China
We linked the cex gene BBa_K118022 into a pET30a(+) backbone, then transformed this plasmid into BL21. After the overnight (about 15h) induction under the condition of 16℃ 0.08 mM, 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) assey. The result is shown on Fig.1.