Coding

Part:BBa_K4388001

Designed by: Venkata Kalyan Ghadiyaram   Group: iGEM22_KCL_UK   (2022-09-28)


Rhodotorula glutinis Tyrosine Ammonia-Lyase Mutant (RgTAL S9N/A11T/E518V)

The Rhodotorula glutinis tyrosine ammonia-lyase (RgTAL) is an enzyme that catalyses the deamination of L-tyrosine to produce p-coumaric acid.

TAL 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 optimised for E. coli K12, mutated at the desired positions, and two stop codons (TAATAA) were added to terminate translation.

This part has been successfully ligated into a transcriptional unit: https://parts.igem.org/Part:BBa_K4388005:Experience


Zhou, S., Liu, P., Chen, J., Du, G., Li, H. et Zhou, J.. (2016). Characterization of mutants of a tyrosine ammonia-lyase from Rhodotorula glutinis. Applied microbiology and biotechnology, 100(24), 10443‑10452. doi:10.1007/s00253-016-7672-8


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2021
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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