Difference between revisions of "Part:BBa K3771002"

Revision as of 01:55, 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 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: cs PCR product (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-P_trc-cs (1272 bp)

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

Fig. 8. Taurine production of P_trc-cdo1 +P_lacI-csad +P_trc-cs in different growth mediums.

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