Difference between revisions of "Part:BBa K118022"
Line 15: Line 15:
 
*'''Summary:''' CMC enzyme activity assay and congo red assay for enzyme activity
 
*'''Summary:''' CMC enzyme activity assay and congo red assay for enzyme activity
  
−
==Characterization from iGEM18-UESTC-China==
+
===Characterization from iGEM18-UESTC-China===
−
===Molecular weight===
+
====Molecular weight====
 
This gene codes for a protein of 485 amino acids with a molecular mass of 51.2 kDa.
 
This gene codes for a protein of 485 amino acids with a molecular mass of 51.2 kDa.
−
===Enzyme digestion===
+
====Enzyme digestion====
−
[[File:T--UESTC-China--Enzyme digestion|500px|thumb|center|'''Fig. 1''' Enzyme activity of the total cellulase]]
+
[[File:T--UESTC-China--Enzyme digestion|500px|thumb|center|'''Fig. 1''' ]]
−
===Enzyme activity ===
+
====Enzyme activity====
 
We constructed a plasmid containing cenA gene [https://parts.igem.org/Part:BBa_K118023 BBa_K118023], cex gene [https://parts.igem.org/Part:BBa_K118022 BBa_K118022]. We transformed this plasmid into BL21(DE3).
 
We constructed a plasmid containing cenA gene [https://parts.igem.org/Part:BBa_K118023 BBa_K118023], cex gene [https://parts.igem.org/Part:BBa_K118022 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. [[File:T--UESTC-China—total cellulase activity|500px|thumb|center|'''Fig. 2''' Enzyme activity of the total cellulase at pH7.0, 40 ℃]]
 
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. [[File:T--UESTC-China—total cellulase activity|500px|thumb|center|'''Fig. 2''' Enzyme activity of the total cellulase at pH7.0, 40 ℃]]
−
 
+
====References====
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 08:48, 17 October 2018

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 524
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 157
    Illegal NgoMIV site found at 530
    Illegal NgoMIV site found at 1032
  • 1000
    INCOMPATIBLE 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: CMC enzyme activity assay and congo red assay for enzyme activity

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

File:T--UESTC-China--Enzyme digestion
Fig. 1

Enzyme activity

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.
File:T--UESTC-China—total cellulase activity
Fig. 2 Enzyme activity of the total cellulase at pH7.0, 40 ℃

References