A Tryptophan Prenyltransferase with Broad Substrate Tolerance from Bacillus subtilis subsp. natto

Area of Science:

• Microbiology
• Biochemistry
• Synthetic Biology

Background:

• Bacillus subtilis subsp. natto utilizes quorum sensing for biofilm formation, involving the ComX_natto pheromone.
• The ComX_natto pheromone undergoes post-translational modification, specifically farnesylation of its tryptophan residue, forming a unique tricyclic scaffold.

Purpose of the Study:

• To investigate the functional characteristics of ComQ_natto, the enzyme responsible for the farnesylation of ComX_natto.
• To determine the substrate specificity and catalytic mechanism of ComQ_natto in prenylation reactions.

Main Methods:

• In vitro enzymatic assays were performed using ComQ_natto with various ComX_natto analogues and single tryptophan.
• Kinetic parameters for the farnesyl-transfer reaction were calculated to assess enzyme efficiency and substrate recognition.

Main Results:

• ComQ_natto demonstrated the ability to recognize and farnesylate not only full-length ComX_natto but also truncated analogues and even isolated tryptophan.
• Kinetic analysis revealed that ComQ_natto does not require a leader sequence for substrate binding and modification.

Conclusions:

• ComQ_natto is a versatile enzyme with broad substrate tolerance for farnesylation.
• The enzyme's unique properties make it a valuable tool for synthesizing a wide range of prenylated tryptophan derivatives for potential biotechnological applications.