4LU6

Thermostabilized RebH

Summary for 4LU6

• Entry DOI: 10.2210/pdb4lu6/pdb
• Descriptor: Flavin-dependent tryptophan halogenase RebH, PHOSPHATE ION (3 entities in total)
• Functional Keywords: themrostability, halogenase, oxidoreductase
• Biological source: Lechevalieria aerocolonigenes (Nocardia aerocolonigenes)
• Total number of polymer chains: 2
• Total formula weight: 125114.13

Authors

Poor, C.B.,Lewis, J.C. (deposition date: 2013-07-24, release date: 2014-06-11, Last modification date: 2024-02-28)

Primary citation

Poor, C.B.,Andorfer, M.C.,Lewis, J.C.
Improving the Stability and Catalyst Lifetime of the Halogenase RebH By Directed Evolution.
Chembiochem, 15:1286-1289, 2014

PubMed Abstract: We previously reported that the halogenase RebH catalyzes selective halogenation of several heterocycles and carbocycles, but product yields were limited by enzyme instability. Here, we use directed evolution to engineer an RebH variant, 3-LR, with a Topt over 5 °C higher than that of wild-type, and 3-LSR, with a Tm 18 °C higher than that of wild-type. These enzymes provided significantly improved conversion (up to fourfold) for halogenation of tryptophan and several non-natural substrates. This initial evolution of RebH not only provides improved enzymes for immediate synthetic applications, but also establishes a robust protocol for further halogenase evolution.

PubMed: 24849696
DOI: 10.1002/cbic.201300780

Experimental method

X-RAY DIFFRACTION (3.05 Å)