Difference between revisions of "Part:BBa K3771012"
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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 45.85 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<sup>[1]</sup>. | + | <br>L-cysteine sulfonic acid synthase (CS, L-cysteate synthase) is an enzyme consisting of 419 amino acids, and its size is about 45.85 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<sup>[1]</sup>. |
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| − | <img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width: | + | <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> | + | <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> | <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. | + | <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/f/fb/T--NCKU_Tainan--CS_PCR.png" style="width:35%;"> | <img src="https://2021.igem.org/wiki/images/f/fb/T--NCKU_Tainan--CS_PCR.png" style="width:35%;"> | ||
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| − | <p align="center">Fig.2. The electrophoresis result of | + | <p align="center">Fig. 2. The electrophoresis result of <i>cs</i> fragment from PCR. M: Marker; Lane 1-3: <i>cs-6xHis</i> PCR product (1312 bp). |
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<br><b style="font-size:1.3rem">References</b> | <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. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093 | + | <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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Latest revision as of 11:39, 18 October 2021
CS-6xHis
Description
L-cysteine sulfonic acid synthase (CS, L-cysteate synthase) is an enzyme consisting of 419 amino acids, and its size is about 45.85 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 1-3: cs-6xHis PCR product (1312 bp).
In order to use anti-polyhistidine-tag antibodies to detect the production of CS by western blot, we add 6xHis-tag at the C-terminal of CS. Combining CS and CSAD, we are able to produce taurine from O-phospho-L-serine.
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