Difference between revisions of "Part:BBa K3771004"
 
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<br><b style="font-size:1.3rem">Description
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<br><b style="font-size:1.3rem">Description</b>
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<br>L-cysteine dioxygenase (CDO1) is an enzyme that weighs 23 kDa. CDO1 functions in the L-cysteine sulfinic acid taurine biosynthesis pathway, converting L-cysteine to L-cysteine sulfinic acid. [1]
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<br>L-cysteine dioxygenase (CDO1) is an enzyme that weighs 22 kDa. CDO1 functions in the L-cysteine sulfinic acid taurine biosynthesis pathway, converting L-cysteine to L-cysteine sulfinic acid. [1]
 
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<br><b style="font-size:1.3rem">Biology</b>
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<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:50%;"></html>
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<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:60%;">
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  <p align="center">Fig. 1. Taurine pathways in <i>E. coli</i></p>
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<br>CDO1 is part of the L-cysteine sulfinic acid pathway, one of three possible taurine synthesis pathways. CDO1 oxidizes L-cysteine naturally found in the intestine into L-cysteine sulfinic acid, which is later converted to taurine by CSAD.[1]
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===Usage and Biology===
 
===Usage and Biology===
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<br><b style="font-size:1.3rem">Usage
 
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<br>CDO1 was used in in vivo testing of taurine production. The sequence for CDO1 enzyme and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).  
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<br>CDO1 was used in in vivo testing of taurine production. The sequence for CDO1 enzyme and <i>trc</i> promoter were ligated and transformed into <i>E. coli</i> to calculate taurine production using high-performance liquid chromatography (HPLC).  
 
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<br><b style="font-size:1.3rem">Characterization  
 
<br><b style="font-size:1.3rem">Characterization  
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<br>The CDO1 fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.<br>
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<br>The <i>cdo1</i> fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.<br>
  
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<p>Fig. 1 Confirmation of cdo1 fragment by PCR. M: Marker; Lane 1: cdo1 (603 bp)</p>
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<p align="center">Fig. 2. Confirmation of <i>cdo1</i> fragment by PCR. M: Marker; Lane 1: <i>cdo1</i> (603 bp)</p>
 
    
 
    
 
    
 
    
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<p>Fig. 2 Confirmation of  pSB4KI-Ptrc-CDO1 by digestion.M: Marker; Lane 1~3: Different colonies of pSB4KI-Ptrc-CDO1 (5470 bp)</p>
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<p align="center">Fig. 3. Confirmation of  pSB4KI-<i>P<sub>trc</sub>-cdo1</i> by digestion.M: Marker; Lane 1~3: Different colonies of pSB4KI-<i>P<sub>trc</sub>-cdo1</i> (5470 bp)</p>
 
    
 
    
 
    
 
    
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  <p>Fig. 3 Confirmation of protein expression of CDO1.M: Marker; Lane1: CDO1 in DH5α without induction; Lane2: CDO1 in DH5α with induction (~22 kDa)</p>
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  <p align="center">Fig. 4. Confirmation of protein expression of CDO1. M: Marker; Lane1: CDO1 in DH5α without induction; Lane2: CDO1 in DH5α with induction (~22 kDa)</p>
 
    
 
    
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<p>Fig. 4 Taurine production of Ptrc-cdo1 +PlacI-csad +Ptrc-cs in different growth mediums.</p>
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<br><b style="font-size:1.3rem">References
 
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<br>Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093<br>
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<br>1. Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093<br>
  
 
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Latest revision as of 03:53, 22 October 2021


CDO1


Description


L-cysteine dioxygenase (CDO1) is an enzyme that weighs 22 kDa. CDO1 functions in the L-cysteine sulfinic acid taurine biosynthesis pathway, converting L-cysteine to L-cysteine sulfinic acid. [1]

Biology

Fig. 1. Taurine pathways in E. coli


CDO1 is part of the L-cysteine sulfinic acid pathway, one of three possible taurine synthesis pathways. CDO1 oxidizes L-cysteine naturally found in the intestine into L-cysteine sulfinic acid, which is later converted to taurine by CSAD.[1]

Usage

CDO1 was used in in vivo testing of taurine production. The sequence for CDO1 enzyme and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).



Characterization

The cdo1 fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.

Fig. 2. Confirmation of cdo1 fragment by PCR. M: Marker; Lane 1: cdo1 (603 bp)


Fig. 3. Confirmation of pSB4KI-Ptrc-cdo1 by digestion.M: Marker; Lane 1~3: Different colonies of pSB4KI-Ptrc-cdo1 (5470 bp)


Fig. 4. Confirmation of protein expression of CDO1. M: Marker; Lane1: CDO1 in DH5α without induction; Lane2: CDO1 in DH5α with induction (~22 kDa)




References

1. Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 447
  • 1000
    COMPATIBLE WITH RFC[1000]