Difference between revisions of "Part:BBa K4621000"
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[[Image:https://static.igem.wiki/teams/4621/wiki/parts/pathways-for-5-hydroxyectoine-synthesis.jpg|thumbnail|750px|center|'''Figure 1:''' | [[Image:https://static.igem.wiki/teams/4621/wiki/parts/pathways-for-5-hydroxyectoine-synthesis.jpg|thumbnail|750px|center|'''Figure 1:''' |
Revision as of 04:56, 2 October 2023
EctOperon is a cluster of genes encoding four enzymes that synthesize 5-Hydroxyectoine
Usage, Biology and Characterization:
EctOperon contains ectA, ectB, ectC, and ectD.
ectB is the coding sequence of L-2, 4-diaminobutyrate --2-oxoglutarate aminotransferase, L-Aspartate 4-semialdehyde can be converted to L-2,4-diaminobutanoate. ectA is the coding sequence for L-2,4-diaminobutyric acid acetyltransferase, L-2, 4-diaminobutanoate can be converted to N4-acetyl-L-2,4-diaminobutyrate. ectC is the coding sequence of L-ectoine synthase, which can convert N4-acetyl-L-2,4-diaminobutyrate to L-Ectoine. These three genes encode enzymes Ectoine: (4S)-2-methyl-1,4,5, 6-tetrahydropyrim idine-4-carboxylic acid, located on the same operon, forming the ectABC gene cluster. The Ectoine is then activated by ectoine hydroxylase, which is encoded by the ectD gene, to generate 5-Hydroxyectoine. The synthesis of 5-Hydroxyectoine from L-Aspartate occurs naturally in some moderately halophilic microorganisms, as shown in Figure 1.
Streptomyces SCUT-3 was modified to overexpressed EctOperon. They were then seeded in a 40 mL LB medium system. After fermentation at 40 °C and 220 rpm for 72h, the product 5-Hydroxyectoine was determined as follows.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 3227
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 3227
Illegal NotI site found at 571
Illegal NotI site found at 1456
Illegal NotI site found at 1774 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 753
Illegal XhoI site found at 927 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 3227
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 3227
Illegal NgoMIV site found at 788
Illegal NgoMIV site found at 1194
Illegal NgoMIV site found at 1200
Illegal NgoMIV site found at 1491
Illegal NgoMIV site found at 2540
Illegal NgoMIV site found at 2732
Illegal NgoMIV site found at 3242 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 376
Illegal BsaI.rc site found at 556
Illegal BsaI.rc site found at 1338
Illegal BsaI.rc site found at 1866
Illegal BsaI.rc site found at 2121
Illegal BsaI.rc site found at 2211
Illegal BsaI.rc site found at 2286
Illegal SapI site found at 414