Difference between revisions of "Part:BBa K3771002"
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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 | + | <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/6/67/T--NCKU_Tainan--cs_pcr%28without_6xHis%29.gif" style="width:35%;"> | <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> | + | <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/9/9b/T--NCKU_Tainan--CS-colony-PCR.png" style="width:35%;"> | <img src="https://2021.igem.org/wiki/images/9/9b/T--NCKU_Tainan--CS-colony-PCR.png" style="width:35%;"> | ||
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− | <p align="center">Fig. 5. Confirmation of cs fragment by colony PCR. | + | <p align="center">Fig. 5. Confirmation of <i>cs</i> fragment by colony PCR. |
− | M: Marker; Lane 1~6: Different colonies of pSUI- | + | M: Marker; Lane 1~6: Different colonies of pSUI-<i>P<sub>trc</sub>-cs</i> (1272 bp) |
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<img src="https://2021.igem.org/wiki/images/1/1d/T--NCKU_Tainan--CS-digestion.png" style="width:35%;"> | <img src="https://2021.igem.org/wiki/images/1/1d/T--NCKU_Tainan--CS-digestion.png" style="width:35%;"> | ||
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− | <p align="center">Fig. 6. Confirmation of pSUI- | + | <p align="center">Fig. 6. Confirmation of pSUI-<i>P<sub>trc</sub>-cs</i> by digestion. |
− | M: Marker; Lane 1: Colony of pSUI-<i>P<sub>trc</sub>- | + | M: Marker; Lane 1: Colony of pSUI-<i>P<sub>trc</sub>-cs</i></p> |
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<img src="https://2021.igem.org/wiki/images/6/64/T--NCKU_Tainan--CS-PAGE%28DH5a%29.png" style="width:35%;"> | <img src="https://2021.igem.org/wiki/images/6/64/T--NCKU_Tainan--CS-PAGE%28DH5a%29.png" style="width:35%;"> | ||
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− | <p align="center">Fig. 7. Confirmation of the protein expression of CS | + | <p align="center">Fig. 7. Confirmation of the protein expression of CS. |
M: Marker; Lane 1: CS in DH5α with induction (~46 kDa)</p> | M: Marker; Lane 1: CS in DH5α with induction (~46 kDa)</p> | ||
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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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 493