Part:BBa_K2423001
CsADH-2946 under the control of a constitutive promoter (BBa_J23106)
This part codes for aldehyde dehydrogenase 2946, which catalyses the conversion of crocetin dialdehyde into crocetin, using NAD+ as a cofactor, in Crocus sativus. It contains a constitutive promoter, as well as a His tag for easy purification. A version of this biobrick containing an inducible promoter is BBa_K2423007.
Usage and Biology
Saffron, a well recognized, but expensive spice has not only uses in terms of cooking but compounds found in saffron have been shown to help with inflammation (1), neurodegenerative diseases (2) and more. Some of those compounds namely zeaxanthin, crocetin dialdehyde, crocetin and crocin are all a part of the same metabolic pathway in the plant specie Crocus Sativus. Not only are these compounds in saffron helpful in terms their medicinal properties, but also the fact that they are very colorful. These aspects was what drew us at iGEM Uppsala 2017 to work with the pathway from zeaxanthin to crocin in the BioBrick format, put also to integrate the metabolic steps that leads up to crocin (the pathway from farnesyl pyrophospate (FPP) to zeaxanthin) on the chromosome of Escherichia Coli. The enzyme presented on this page catalyzes the second reaction in the zeaxanthin to crocin pathway.
CsADH2946 was discovered through transcriptomic analysis of the chromoplasts of Crocus Sativus (3). The active site of CsADH2946 can be found around a loop containing three cystenin residues in a row (C337, C338, C339; positions were determined from our homology model). The residues that are conserved were found by looking at the template (PDB: 4fqf) (4) used in our homology model.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 195
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 602
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1119
Illegal SapI.rc site found at 227
References
1. Papandreou MA, Kanakis CD, Polissiou MG, Efthimiopoulos S, Cordopatis P, Margarity M, et al. Inhibitory Activity on Amyloid-β Aggregation and Antioxidant Properties of Crocus sativus Stigmas Extract and Its Crocin Constituents. J Agric Food Chem. 2006 Nov 1;54(23):8762–8.
2. Chen L, Qi Y, Yang X. Neuroprotective effects of crocin against oxidative stress induced by ischemia/reperfusion injury in rat retina. Ophthalmic Res. 2015;54(3):157–68.
3. Gómez-Gómez L, Parra-Vega V, Rivas-Sendra A, Seguí-Simarro JM, Molina RV, Pallotti C, et al. Unraveling Massive Crocins Transport and Accumulation through Proteome and Microscopy Tools during the Development of Saffron Stigma. Int J Mol Sci [Internet]. 2017 Jan 1;18(1). Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5297711/
4. Lang BS, Gorren ACF, Oberdorfer G, Wenzl MV, Furdui CM, Poole LB, et al. Vascular Bioactivation of Nitroglycerin by Aldehyde Dehydrogenase-2. J Biol Chem. 2012 Nov 2;287(45):38124–34.
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