Journal
CHEMBIOCHEM
Volume 4, Issue 1, Pages 34-39Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.200390011
Keywords
asymmetric catalysis; directed evolution; enzymes; mutagenesis; oligonucleotides
Ask authors/readers for more resources
A new and practical method for gene recombination with formation of libraries of mutant genes is presented. The method is based on the assembly of appropriately prepared oligonucleotides whose design is guided by sequence information. High recombination frequency with formation of full-length products is achieved by controlled overlapping of the designed oligomers. This process (ADO) minimizes self-hybridization of parental genes, which constitutes a significant advantage over conventional family shuffling as used in the directed evolution of functional enzymes. ADO was applied to the recombination of two lipase family genes from Bacillus subtilis (LipA and LipB). In a library of 3000 lipase variants created by this method, several were found that display increased enantioselectivity in a model reaction involving the hydrolysis of a meso-diacetate.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available