BACKGROUND: Directed evolution by DNA shuffling has been used to modify physical and catalytic properties of biological systems. We have shuffled two highly homologous triazine hydrolases and conducted an exploration of the substrate specificities of the resulting enzymes to acquire a better understanding of the possible distributions of novel functions in sequence space. RESULTS: Both parental enzymes and a library of 1600 variant triazine hydrolases were screened against a synthetic library of 15 triazines. The shuffled library contained enzymes with up to 150-fold greater transformation rates than either parent. It also contained enzymes that hydrolyzed five of eight triazines that were not substrates for either starting enzyme. CONCLUSIONS: Permutation of nine amino acid differences resulted in a set of enzymes with surprisingly diverse patterns of reactions catalyzed. The functional richness of this small area of sequence space may aid our understanding of both natural and artificial evolution.
BACKGROUND: Directed evolution by DNA shuffling has been used to modify physical and catalytic properties of biological systems. We have shuffled two highly homologous triazine hydrolases and conducted an exploration of the substrate specificities of the resulting enzymes to acquire a better understanding of the possible distributions of novel functions in sequence space. RESULTS: Both parental enzymes and a library of 1600 variant triazine hydrolases were screened against a synthetic library of 15 triazines. The shuffled library contained enzymes with up to 150-fold greater transformation rates than either parent. It also contained enzymes that hydrolyzed five of eight triazines that were not substrates for either starting enzyme. CONCLUSIONS: Permutation of nine amino acid differences resulted in a set of enzymes with surprisingly diverse patterns of reactions catalyzed. The functional richness of this small area of sequence space may aid our understanding of both natural and artificial evolution.
Authors: Sahil Balotra; Janet Newman; Nathan P Cowieson; Nigel G French; Peter M Campbell; Lyndall J Briggs; Andrew C Warden; Christopher J Easton; Thomas S Peat; Colin Scott Journal: Appl Environ Microbiol Date: 2014-10-31 Impact factor: 4.792
Authors: Yasuaki Kawarasaki; Karl E Griswold; James D Stevenson; Tzvia Selzer; Stephen J Benkovic; Brent L Iverson; George Georgiou Journal: Nucleic Acids Res Date: 2003-11-01 Impact factor: 16.971
Authors: Colin Scott; Colin J Jackson; Chris W Coppin; Roslyn G Mourant; Margaret E Hilton; Tara D Sutherland; Robyn J Russell; John G Oakeshott Journal: Appl Environ Microbiol Date: 2009-02-06 Impact factor: 4.792
Authors: Jennifer L Seffernick; Nir Shapir; Michael Schoeb; Gilbert Johnson; Michael J Sadowsky; Lawrence P Wackett Journal: Appl Environ Microbiol Date: 2002-09 Impact factor: 4.792