Difference between revisions of "Part:BBa K3771003"

Revision as of 19:11, 19 October 2021 (view source) Evelynnn (Talk | contribs) ← Older edit		Revision as of 11:22, 20 October 2021 (view source) ShaneLee (Talk | contribs) Newer edit →
Line 33:		Line 33:
	<br>The CSAD fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.		<br>The CSAD fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.
	<div style="width=100%; display:flex; align-items: center; justify-content: center;">		<div style="width=100%; display:flex; align-items: center; justify-content: center;">
−	<img src="https://2021.igem.org/wiki/images/f/fd/T--NCKU_Tainan--csad_his.jpg	+	<img src="https://2021.igem.org/wiki/images/8/86/T--NCKU_Tainan--CSAD-PCR.png
	" style="width:35%;">		" style="width:35%;">
	</div>		</div>

---

Revision as of 11:22, 20 October 2021

CSAD

Description

L-Cysteine sulfinic acid decarboxylase (CSAD) is an enzyme consisting of 493 amino acids and weighs 55.4 kDa. CSAD functions in the taurine biosynthesis pathway, converting L-Cysteine to taurine [1].


Biology

Fig. 1. Taurine production pathway

CSAD is part of the L-cysteine sulfinic acid pathway, one of two possible taurine synthesis pathways. CSAD catalyzes the decarboxylation of L-Cysteine sulfinic acid into hypotaurine, which is spontaneously oxidized to taurine [1].


Usage

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


Characterization

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

Fig. 2. PCR of csad sequence (1500 bp).

SDS-PAGE and western blot of CSAD enzyme to confirm protein expression.

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