Part:BBa_K4339000
Non-polar amino acid transporter (CycA)
CycA is an inner membrane permease [1], which can facilitate the uptake of non-polar amino acids (including D-alanine, D-serine, glycine and L-alanine) via a proton symport mechanism [2]. CycA was first identified in the K12 strain of E. coli.
CycA was codon-optimised for expression in E. coli BL21 DE3 strains and their derivatives. CycA was fused with the constitutive promoter PJ23100 and a C-terminus His tag and engineered into the strain BL21 DE3 pLysS. CycA was engineered into this strain with an aim to increase uptake of L-alanine from an alanine-doped LB growth medium, to facilitate synthesis of alanine-rich proteins such as MaSps (Major Ampullate Silk proteins).
Usage and Biology
Sequence and Features
The sequence originates from E. coli MG1655 K-12 (Hook et al. 2022)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 340
- 1000COMPATIBLE WITH RFC[1000]
References
[1] - Hook C et al. The Escherichia coli Amino Acid Uptake Protein CycA: Regulation of Its Synthesis and Practical Application in l-Isoleucine Production. Microorganisms. 2022;10(3):647. doi: https://doi.org/10.3390/microorganisms10030647
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