Difference between revisions of "Team:Duke/Basic Part"
| Line 4: | Line 4: | ||
<h1>BAPT</h1> | <h1>BAPT</h1> | ||
<p><br> | <p><br> | ||
| − | <h4><center> | + | <h4><center>Name: </h4></center><br> |
Phenylpropanoyltransferase (BAPT, TAX7) | Phenylpropanoyltransferase (BAPT, TAX7) | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>Reaction: </h4></center><br> | |
baccatin III + (3R)-3-amino-3-phenylpropanoyl-CoA → N-debenzoyl-(3'-RS)-2'-deoxytaxol + coenzyme A | baccatin III + (3R)-3-amino-3-phenylpropanoyl-CoA → N-debenzoyl-(3'-RS)-2'-deoxytaxol + coenzyme A | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>Km: </h4></center><br> | |
5.6 uM for (3R)-3-amino-3-phenylpropanoyl-CoA; | 5.6 uM for (3R)-3-amino-3-phenylpropanoyl-CoA; | ||
68 uM for baccatin III; | 68 uM for baccatin III; | ||
| Line 18: | Line 18: | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>Kcat: </h4></center><br> | |
0.0583 ± 0.0010 s<sup>-1</sup> (Thornburg, 2015) | 0.0583 ± 0.0010 s<sup>-1</sup> (Thornburg, 2015) | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>Enzyme Description: </h4></center><br> | |
BAPT is the enzyme that adds the C13 Taxol side chain partially formed to the Taxol backbone. It makes use of <br> | BAPT is the enzyme that adds the C13 Taxol side chain partially formed to the Taxol backbone. It makes use of <br> | ||
CoA chemistry to add the side chain to the 13’ hydroxyl group of baccatin III. | CoA chemistry to add the side chain to the 13’ hydroxyl group of baccatin III. | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>Sequence</h4></center><br> | |
atgaagaagactgggagtttcgcggagttccacgttaatatgatcgagcgcgtgatggttcgtccttgccttccttcacctaagactattttgcctttgt | atgaagaagactgggagtttcgcggagttccacgttaatatgatcgagcgcgtgatggttcgtccttgccttccttcacctaagactattttgcctttgt | ||
ctgcgatcgataacatggcacgtgccttcagtaatgtcttactggtgtacgccgccaacatggatcgcgtcagcgcggacccggcaaaggttattcgtga | ctgcgatcgataacatggcacgtgccttcagtaatgtcttactggtgtacgccgccaacatggatcgcgtcagcgcggacccggcaaaggttattcgtga | ||
| Line 44: | Line 44: | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>Design Notes</h4></center><br> | |
Part digested from vector designed by Duke's 2016 team. Original vector constructed by Gibson Assembly of BAPT Gblock into pSMART-HC-Amp vector. | Part digested from vector designed by Duke's 2016 team. Original vector constructed by Gibson Assembly of BAPT Gblock into pSMART-HC-Amp vector. | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>Source</h4></center><br> | |
Origin from Pacific Yew Tree, Taxus brevifolia | Origin from Pacific Yew Tree, Taxus brevifolia | ||
<br><br> | <br><br> | ||
| − | + | <h4><center>References</h4></center><br> | |
Thornburg, C. (2015). Development of a four-step semi-biosynthesis of the anticancer drug paclitaxel and its analogues (Doctoral dissertation, Michigan State University). | Thornburg, C. (2015). Development of a four-step semi-biosynthesis of the anticancer drug paclitaxel and its analogues (Doctoral dissertation, Michigan State University). | ||
<br> </p> | <br> </p> | ||
Revision as of 20:03, 17 October 2018
BAPT
Name:
Phenylpropanoyltransferase (BAPT, TAX7)
Reaction:
baccatin III + (3R)-3-amino-3-phenylpropanoyl-CoA → N-debenzoyl-(3'-RS)-2'-deoxytaxol + coenzyme A
Km:
5.6 uM for (3R)-3-amino-3-phenylpropanoyl-CoA; 68 uM for baccatin III; 2.4 uM for baccatin III and 4.9 uM beta-phenylalanoyl-CoA (Thornburg, 2015)
Kcat:
0.0583 ± 0.0010 s-1 (Thornburg, 2015)
Enzyme Description:
BAPT is the enzyme that adds the C13 Taxol side chain partially formed to the Taxol backbone. It makes use of
CoA chemistry to add the side chain to the 13’ hydroxyl group of baccatin III.
Sequence
atgaagaagactgggagtttcgcggagttccacgttaatatgatcgagcgcgtgatggttcgtccttgccttccttcacctaagactattttgcctttgt ctgcgatcgataacatggcacgtgccttcagtaatgtcttactggtgtacgccgccaacatggatcgcgtcagcgcggacccggcaaaggttattcgtga ggcgcttagcaaggtattggtctactattacccttttgctggccgcttgcgtaacaaagaaaatggagagcttgaagtggagtgcacaggacaaggagta ttgtttttagaagccatggctgacagcgacctttcagtattaactgacctggacaactacaatcctagttttcagcagcttatctttagcttgccgcaag ataccgacatcgaggatttgcatttgttgatcgtacaggtcacacgcttcacatgtggtgggtttgttgtcggtgctaacgtctatggttccgcctgtga cgcaaaaggattcggtcaattcttacagtctatggcggagatggctcgcggagaagtaaagccctccatcgaaccaatctggaatcgcgaattggttaaa cttgagcactgtatgccttttcgtatgtcacacttgcagattattcacgcgccagtcattgaggagaagtttgtccagacatcgcttgtgatcaacttcg aaatcatcaaccatattcgtcgtcgcatcatggaagagcgtaaagagagcctgtcttctttcgaaattgttgccgcgttagtctggcttgcgaagattaa ggcgtttcaaattccacattccgaaaatgtgaaactgctgttcgccatggacctgcgccgctctttcaatccgccgctgccccatgggtattatgggaat gcatttgggatcgcttgcgcgatggacaacgtgcatgatctgttgtcgggtagcctgttgcgtaccattatgatcattaagaagtctaaattttccctgc ataaagaattaaattctaagacagttatgagtagttccgtagtggatgtaaataccaaatttgaggacgttgtgtcgatctccgactggcgccacagcat ttactacgaagtagactttggatggggagacgcaatgaatgtgagtacgatgcttcagcaacaagaacacgaaaaatcgctgccaacgtattttagtttt ttgcaatcgaccaagaatatgcctgacggtatcaagatgttaatgtttatgcctcctagtaaacttaagaaatttaagattgaaatcgaagccatgatta agaaatatgttacgaaggtatgcccttctaagctttgataa
Design Notes
Part digested from vector designed by Duke's 2016 team. Original vector constructed by Gibson Assembly of BAPT Gblock into pSMART-HC-Amp vector.
Source
Origin from Pacific Yew Tree, Taxus brevifolia
References
Thornburg, C. (2015). Development of a four-step semi-biosynthesis of the anticancer drug paclitaxel and its analogues (Doctoral dissertation, Michigan State University).