Difference between revisions of "Part:BBa K3771002"

 
 
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<partinfo>BBa_K3771002 short</partinfo>
 
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<br><br><b style="font-size:1.3rem">Description</b>
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<br>L-cysteine sulfonic acid synthase (CS, L-cysteate synthase) is an enzyme consisting of 419 amino acids, and its size is about 46 kDa. It catalyzes the conversion of O-phospho-L-serine and sulfite into L-cysteine sulfonic acid (L-cysteate) and phosphate (Fig. 1). Moreover, CS does not catalyze the production of L-cysteine due to its specificity to sulfite [1].
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<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:50%;">
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<p align="center">Fig. 1. L-cysteine sulfonic acid synthase catalyzes the reaction that turns O-phospho-L-serine into L-cysteine sulfonic acid. CDO1, L-cysteine dioxygenase; CSAD, L-cysteine sulfinic acid decarboxylase.</p>
  
 
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<br><b style="font-size:1.3rem">Usage and Biology</b>
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<br>The DNA sequence of CS was synthesized by IDT and was amplified by PCR. The Agarose gel electrophoresis result is shown in Fig. 2. The part has been confirmed by sequencing and has no mutations.
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<img src="https://2021.igem.org/wiki/images/6/67/T--NCKU_Tainan--cs_pcr%28without_6xHis%29.gif" style="width:35%;">
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<p align="center">Fig. 2. The electrophoresis result of <i>cs</i> fragment from PCR. M: Marker; Lane 1: <i>cs</i> (1257 bp).
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<img src="https://2021.igem.org/wiki/images/e/e3/T--NCKU_Tainan--CS-Vactor-digestion.png" style="width:35%;">
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<p align="center">Fig. 3.  Confirmation of pSUI fragment by digestion. M: Marker; Lane 1: pSUI (3664 bp)</p>
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<img src="https://2021.igem.org/wiki/images/3/35/T--NCKU_Tainan--CS-plate%28DH5a%29.png" style="width:35%;">
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<p align="center">Fig. 4. Transformation / CS in DH5α</p>
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<p align="center">Fig. 5. Confirmation of <i>cs</i> fragment by colony PCR.
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  M: Marker; Lane 1~6: Different colonies of pSUI-<i>P<sub>trc</sub>-cs</i> (1272 bp)
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<p align="center">Fig. 6. Confirmation of  pSUI-<i>P<sub>trc</sub>-cs</i> by digestion.
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  M: Marker; Lane 1: Colony of pSUI-<i>P<sub>trc</sub>-cs</i></p>
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<p align="center">Fig. 7. Confirmation of the protein expression of CS.
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M: Marker; Lane 1: CS in DH5α with induction (~46 kDa)</p>
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<br>Taurine production yield of CSAD with other production enzymes calculated by high-performance liquid chromatography (HPLC).<br>
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<br><b style="font-size:1.3rem">References</b>
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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. <i>Journal of Agricultural and Food Chemistry</i>. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3771002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3771002 SequenceAndFeatures</partinfo>

Latest revision as of 03:35, 22 October 2021


CS

Description


L-cysteine sulfonic acid synthase (CS, L-cysteate synthase) is an enzyme consisting of 419 amino acids, and its size is about 46 kDa. It catalyzes the conversion of O-phospho-L-serine and sulfite into L-cysteine sulfonic acid (L-cysteate) and phosphate (Fig. 1). Moreover, CS does not catalyze the production of L-cysteine due to its specificity to sulfite [1].

Fig. 1. L-cysteine sulfonic acid synthase catalyzes the reaction that turns O-phospho-L-serine into L-cysteine sulfonic acid. CDO1, L-cysteine dioxygenase; CSAD, L-cysteine sulfinic acid decarboxylase.



Usage and Biology

The DNA sequence of CS was synthesized by IDT and was amplified by PCR. The Agarose gel electrophoresis result is shown in Fig. 2. The part has been confirmed by sequencing and has no mutations.

Fig. 2. The electrophoresis result of cs fragment from PCR. M: Marker; Lane 1: cs (1257 bp).

Fig. 3. Confirmation of pSUI fragment by digestion. M: Marker; Lane 1: pSUI (3664 bp)


Fig. 4. Transformation / CS in DH5α



Fig. 5. Confirmation of cs fragment by colony PCR. M: Marker; Lane 1~6: Different colonies of pSUI-Ptrc-cs (1272 bp)


Fig. 6. Confirmation of pSUI-Ptrc-cs by digestion. M: Marker; Lane 1: Colony of pSUI-Ptrc-cs


Fig. 7. Confirmation of the protein expression of CS. M: Marker; Lane 1: CS in DH5α with induction (~46 kDa)


Taurine production yield of CSAD with other production enzymes calculated by high-performance liquid chromatography (HPLC).






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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 493