Part:BBa_K5378006:Sequence, Features, and Subparts

Usage and Biology

TynA is a periplasmic amine oxidase containing copper and topaquinone cofactors, which converts amines to aldehydes.TynA-G494S discovered by Rottinghaus, A. G. et al converts phenethylamine (PEA) to phenylacetaldehyde (PAG) specifically.

Functional Verification

How to identify Phenylethylamine (PEA)?

In Escherichia coli, TynA is a periplasmic amine oxidase containing copper and topaquinone cofactors.Aromatic amines such as PEA , Tyramine (Tyra), Dopamine (DA), etc. are oxidized by TynA to the corresponding aldehydes. The study shows that expression of the monoamine oxidase TynA and dehydrogenase FeaB are regulated by the AraC-type FeaR transcription factor, which was shown to activate PtynA in the presence of multiple aldehydes formed from the corresponding amines. Obviously, this sensing system, including tyna-feaR gene cluster, provides an opportunity to sense various aromatic amines.The sensing system converts PEA to phenylacetaldehyde (PAG) by expressing TynA-G494S. When PAG binds to Fear-A81L, the promoter PtynA is activated,starting the expression of metabolic modules downstream.

But it doesn't meet our needs——specific identification of PEA.

In order to better identify PEA, we introduce the FeaR-A81L and TynA-G494S mutant with 580-fold induction and PEA specificity.The variant induces a 580-fold enhancement in expression and is PEA-specific.

Reference

Rottinghaus, A. G., Xi, C., Amrofell, M. B., Yi, H., & Moon, T. S. (2022). Engineering ligand-specific biosensors for aromatic amino acids and neurochemicals. Cell systems, 13(3), 204-214.