Part:BBa_K4588036

Tyrosine aminotransferase (S. cerevisiae), TyrB

This part encodes tyrosine aminotransferase from Escherichia coli K12

Biology

The organism this gene is initially expressed in is Escherichia coli.

TyrB codes for the tyrosine aminotransferase enzyme that carries out the last step in the phenylalanine and tyrosine biosynthetic pathways involving the transamination of keto acids phenylpyruvate and 4-hydroxyphenylpyruvate. Another aminotransferase, coded for by a closely related aspC gene, can also carry out these reactions but has a lower affinity for these substrates and is primarily involved in forming aspartate [1]. The tyrosine repressor regulates TyrB and is crucial in the synthesis of amino acids phenylalanine, tyrosine, aspartate, and leucine [2].

Design

This BioBrick contains a GAL1 promoter (BBa_K2637059) that supports strong inducible expression in the presence of galactose. Following this promoter, the sequence contains a Kozak sequence (BBa_J63003) for optimal ribosome binding. The protein coding sequence, BBa_K4588025, was optimized for S. cerevisiae expression. The coding region also contains an HA tag (BBa_K1150016) for protein detection. The strong ADH1 terminator (BBa_K1486025) completely stops protein expression.

Usage

This enzyme is implemented in the synthesis pathway to produce rosmarinic acid in the S. cerevisiae culture. This enzyme converts 4-hydroxyphenylpyruvate into L-tyrosine by the addition of an amine group and the formation of a double-bonded oxygen on [3].

References

1. Yang, J., & Pittard, J. (1987). Molecular analysis of the regulatory region of the Escherichia coli K-12 tyrB gene. Journal of bacteriology, 169(10), 4710–4715. https://doi.org/10.1128/jb.169.10.4710-4715.1987

2. Fotheringham, I. G., Dacey, S. A., Taylor, P. P., Smith, T. J., Hunter, M. G., Finlay, M. E., Primrose, S. B., Parker, D. M., & Edwards, R. M. (1986). The cloning and sequence analysis of the aspC and tyrB genes from Escherichia coli K12. Comparison of the primary structures of the aspartate aminotransferase and aromatic aminotransferase of E. coli with those of the pig aspartate aminotransferase isoenzymes. The Biochemical journal, 234(3), 593–604. https://doi.org/10.1042/bj2340593

3. Babaei, M., Borja Zamfir, G. M., Chen, X., Christensen, H. B., Kristensen, M., Nielsen, J., & Borodina, I. (2020). Metabolic engineering of saccharomyces cerevisiae for rosmarinic acid production. ACS Synthetic Biology, 9(8), 1978–1988. https://doi.org/10.1021/acssynbio.0c0004

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]