Synthetic pathways and processes for effective production of 5-hydroxytryptophan and serotonin from glucose in Escherichia coli

Background: Tryptophan derivatives such as 5-hydroxytryptophan (5HTP) and serotonin are valuable molecules with pharmaceutical interest. 5HTP is presently mainly obtained by extraction from the plant Griffonia simplicifolia and serotonin is produced by chemical synthesis. A simple biotechnological method for the production of these compounds is desired. Results: In a first attempt to synthesize serotonin from glucose, we used a single engineered Escherichia coli strain and observed a low production of maximal 0.8 +/- 0.2 mg/L of serotonin, probably due to the undesired site-reaction of direct decarboxylation of tryptophan and the consequent decrease of the precursor 5HTP. To circumvent this problem, we have constructed a stepwise system in which the 5HTP production and the serotonin conversion are separated. 962 +/- 58 mg/L of 5HTP was produced in the first step using a recombinant strain with a semi-rationally engineered aromatic amino acid hydroxylase, the highest concentration reported so far. In a subsequent step of 5HTP bioconversion using a recombinant strain harboring a tryptophan decarboxylase, 154.3 +/- 14.3 mg/L of serotonin was produced. Conclusions: We present results of a two-stage fermentation process for the production of 5HTP and serotonin. The first strain is a highly efficient 5HTP producer, and after fermentation the supernatant is separated and used for the production of serotonin. This is the first report for the microbial production of serotonin from glucose.