l-Serine Biosensor-Controlled Fermentative Production of l-Tryptophan Derivatives by Corynebacterium glutamicum

Simple Summary l-tryptophan is an amino acid found in proteins. Its derivatives, such as hydroxylated or halogenated l-tryptophans, find applications in the chemical and pharmaceutical industries, for example, in therapeutic peptides. Biotechnology provides a sustainable way for the production of l-tryptophan and its derivatives. In the final reaction of l-tryptophan biosynthesis in bacteria, such as Corynebacterium glutamicum, another amino acid, l-serine, is incorporated. Here, we show that C. glutamicum TrpB is able to convert indole derivatives, which were added to cells synthesizing l-serine, to the corresponding l-tryptophan derivatives. The gene trpB was expressed under the control of the l-serine-responsive transcriptional activator SerR in the C. glutamicum cells engineered for this fermentation process. l-Tryptophan derivatives, such as hydroxylated or halogenated l-tryptophans, are used in therapeutic peptides and agrochemicals and as precursors of bioactive compounds, such as serotonin. l-Tryptophan biosynthesis depends on another proteinogenic amino acid, l-serine, which is condensed with indole-3-glycerophosphate by tryptophan synthase. This enzyme is composed of the alpha-subunit TrpA, which catalyzes the retro-aldol cleavage of indole-3-glycerol phosphate, yielding glyceraldehyde-3-phosphate and indole, and the beta-subunit TrpB that catalyzes the beta-substitution reaction between indole and l-serine to water and l-tryptophan. TrpA is reported as an allosteric actuator, and its absence severely attenuates TrpB activity. In this study, however, we showed that Corynebacterium glutamicum TrpB is catalytically active in the absence of TrpA. Overexpression of C. glutamicum trpB in a trpBA double deletion mutant supported growth in minimal medium only when exogenously added indole was taken up into the cell and condensed with intracellularly synthesized l-serine. The fluorescence reporter gene of an l-serine biosensor, which was based on the endogenous transcriptional activator SerR and its target promoter P-serE, was replaced by trpB. This allowed for l-serine-dependent expression of trpB in an l-serine-producing strain lacking TrpA. Upon feeding of the respective indole derivatives, this strain produced the l-tryptophan derivatives 5-hydroxytryptophan, 7-bromotryptophan, and 5-fluorotryptophan.

Automated summary

This study demonstrates that TrpB in Corynebacterium glutamicum is catalytically active even in the absence of TrpA, and can convert indole derivatives into corresponding L-tryptophan derivatives. This provides a novel approach for the production of L-tryptophan and its derivatives in biotechnology.