Difference between revisions of "Part:BBa K3633012"
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==Description== | ==Description== | ||
| − | + | [[File:T--Shanghai_SFLS_SPBS--Betalains Synthesis--Pathway.png|600px|center|thumb|Fig 1. Betalains Synthesis Pathway]] | |
Betalains are water-soluble nitrogen-containing pigments that are subdivided in red-violet betacyanins and yellow-orange betaxanthins. Due to glycosylation and acylation betalains exhibit a huge structural diversity. Betanin (betanidin-5-O--glucoside) is the most common betacyanin in the plant kingdom. | Betalains are water-soluble nitrogen-containing pigments that are subdivided in red-violet betacyanins and yellow-orange betaxanthins. Due to glycosylation and acylation betalains exhibit a huge structural diversity. Betanin (betanidin-5-O--glucoside) is the most common betacyanin in the plant kingdom. | ||
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The biosynthesis of betalains in plants excludes that of anthocyanins. During the biosynthesis of betalains in the cytoplasm three enzymes are involved: Tyrosinase, 4,5-DOPA-extradioldioxygenase, and betanidin-glucosyltransferase. The amino acid L-tyrosine, which is enzymatically formed over the shikimate pathway from arogenic acid, is the precursor for the biosynthesis of L-DOPA. Tyrosine is hydroxylated by means of the enzyme tyrosinase to DOPA (I) that is formed to betalamic acid or to cyclo-DOPA. The biosynthesis of betalamic acid, which is the basic structure of betalains as follow: 4,5-DOPA-extradiol dioxygenase opens the cyclic ring of L-DOPA between carbons 4 and 5, thus producing 4,5-seco-DOPA (II). This intermediate product occurs naturally. Due to spontaneous intramolecular condensation between the amine group and the aldehyde group of 4,5-seco-DOPA betalamic acid is formed. | The biosynthesis of betalains in plants excludes that of anthocyanins. During the biosynthesis of betalains in the cytoplasm three enzymes are involved: Tyrosinase, 4,5-DOPA-extradioldioxygenase, and betanidin-glucosyltransferase. The amino acid L-tyrosine, which is enzymatically formed over the shikimate pathway from arogenic acid, is the precursor for the biosynthesis of L-DOPA. Tyrosine is hydroxylated by means of the enzyme tyrosinase to DOPA (I) that is formed to betalamic acid or to cyclo-DOPA. The biosynthesis of betalamic acid, which is the basic structure of betalains as follow: 4,5-DOPA-extradiol dioxygenase opens the cyclic ring of L-DOPA between carbons 4 and 5, thus producing 4,5-seco-DOPA (II). This intermediate product occurs naturally. Due to spontaneous intramolecular condensation between the amine group and the aldehyde group of 4,5-seco-DOPA betalamic acid is formed. | ||
| − | In order to make the production of the Betacyanin | + | In order to make the production of the Betacyanin, Shanghai_SFLS_SPBS built the biobrick with 4,5-DODA and T7 promoter and added the substrate L-Dopa and 0.1mM IPTG to induce the promoter. The 4,5-seco-DOPA will spontaneously convert into Betalamic acid with the help of ascorbic acid(Vitamin C).And Dopaxanthin/Indoline-Betacyanin will be subsequently synthesized by adding the substrate of L-DOPA/Indoline. The biobrick was successfully expressed in both E.coli BL21(DE3) and vibrio natriegens. The two kind of pigments were produced and application of the hair-dye process was successful as well. |
==Experiments & Results== | ==Experiments & Results== | ||
Revision as of 09:44, 24 October 2020
A composite part to express 4,5-DODA induced by IPTG, responsible for production of betalains
Description
Betalains are water-soluble nitrogen-containing pigments that are subdivided in red-violet betacyanins and yellow-orange betaxanthins. Due to glycosylation and acylation betalains exhibit a huge structural diversity. Betanin (betanidin-5-O--glucoside) is the most common betacyanin in the plant kingdom.
The biosynthesis of betalains in plants excludes that of anthocyanins. During the biosynthesis of betalains in the cytoplasm three enzymes are involved: Tyrosinase, 4,5-DOPA-extradioldioxygenase, and betanidin-glucosyltransferase. The amino acid L-tyrosine, which is enzymatically formed over the shikimate pathway from arogenic acid, is the precursor for the biosynthesis of L-DOPA. Tyrosine is hydroxylated by means of the enzyme tyrosinase to DOPA (I) that is formed to betalamic acid or to cyclo-DOPA. The biosynthesis of betalamic acid, which is the basic structure of betalains as follow: 4,5-DOPA-extradiol dioxygenase opens the cyclic ring of L-DOPA between carbons 4 and 5, thus producing 4,5-seco-DOPA (II). This intermediate product occurs naturally. Due to spontaneous intramolecular condensation between the amine group and the aldehyde group of 4,5-seco-DOPA betalamic acid is formed.
In order to make the production of the Betacyanin, Shanghai_SFLS_SPBS built the biobrick with 4,5-DODA and T7 promoter and added the substrate L-Dopa and 0.1mM IPTG to induce the promoter. The 4,5-seco-DOPA will spontaneously convert into Betalamic acid with the help of ascorbic acid(Vitamin C).And Dopaxanthin/Indoline-Betacyanin will be subsequently synthesized by adding the substrate of L-DOPA/Indoline. The biobrick was successfully expressed in both E.coli BL21(DE3) and vibrio natriegens. The two kind of pigments were produced and application of the hair-dye process was successful as well.
Experiments & Results
Sequence & Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]