Difference between revisions of "Part:BBa K4388001"
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Tyrosine ammonia-lyase is the first enzyme of the general phenylpropanoid pathway that has been used for the production of multiple polyphenols including naringenin, resveratrol, and pterostilbene. This version of the enzyme is a mutant with amino acid changes S9N, A11T, and E518V. These mutations have been shown to increase kcat/km specificity constant from 298 (wild-type) to 391 mM^-1*s^-1 (Zhou et al., 2016). | Tyrosine ammonia-lyase is the first enzyme of the general phenylpropanoid pathway that has been used for the production of multiple polyphenols including naringenin, resveratrol, and pterostilbene. This version of the enzyme is a mutant with amino acid changes S9N, A11T, and E518V. These mutations have been shown to increase kcat/km specificity constant from 298 (wild-type) to 391 mM^-1*s^-1 (Zhou et al., 2016). | ||
| − | The ORF of the wild-type version was obtained from NCBI GenBank code: KF765779.1. This sequence was then codon optimized for E.coli K12, mutated at the desired positions, and two stop codons (TAATAA) were added to terminate translation. | + | The ORF of the wild-type version was obtained from NCBI GenBank code: KF765779.1. This sequence was then codon optimized for E. coli K12, mutated at the desired positions, and two stop codons (TAATAA) were added to terminate translation. |
Revision as of 14:31, 4 October 2022
Rhodotorula glutinis Tyrosine Ammonia-Lyase Mutant (RgTAL S9N/A11T/E518V)
The Rhodotorula glutinis tyrosine ammonia-lyase is an enzyme that catalyses the deamination of L-tyrosine to produce p-coumaric acid.
Tyrosine ammonia-lyase is the first enzyme of the general phenylpropanoid pathway that has been used for the production of multiple polyphenols including naringenin, resveratrol, and pterostilbene. This version of the enzyme is a mutant with amino acid changes S9N, A11T, and E518V. These mutations have been shown to increase kcat/km specificity constant from 298 (wild-type) to 391 mM^-1*s^-1 (Zhou et al., 2016).
The ORF of the wild-type version was obtained from NCBI GenBank code: KF765779.1. This sequence was then codon optimized for E. coli K12, mutated at the desired positions, and two stop codons (TAATAA) were added to terminate translation.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2021
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]