Molecular cloning and functional analysis of serotonin N-acetyltransferase from the cyanobacterium Synechocystis sp PCC 6803

Serotonin N-acetyltransferase (SNAT) catalyzes conversion of serotonin into N-acetylserotonin, which is a direct precursor for melatonin biosynthesis in all organisms. Molecular cloning of plant SNAT from rice led to a screening for SNAT homolog genes in other species. We identified a cyanobacterium SNAT-like gene (cSNAT) that showed 56% amino acid homology with the rice SNAT. To confirm whether cSNAT encoded SNAT enzyme activity, we expressed cSNAT DNA in Escherichia coli and purified the cSNAT protein as a C-terminal His-tagged form. The purified cSNAT protein exhibited SNAT enzyme activities, transferring the acetyl group into either serotonin or tryptamine substrates. The optimum temperature was 55 degrees C, but it was still highly active at 70 degrees C, suggesting that cSNAT is a thermotolerant enzyme. The K-m and V-max were 823m and 1.6nmol/min/mg protein, respectively. The cSNAT gene is highly conserved in all cyanobacterial taxa and seems to be an origin of SNAT in higher plants. The thermotolerance of cSNAT suggests that melatonin plays a role in the response to high-temperature stress. Further analysis of this role of melatonin in higher plants is needed.