New strategies of protein engineering——directed evolution of enzyme in vitro

New strategies of protein engineering—directed evolution of enzymein vitro

New and highly effective strategies for directed enzyme evolution in vitro have been developed in the protein engineering field. They allow engineering all kinds of enzymes in vitro so that new ones with novel functions and features can be obtained by the methods of error-prone PCR, DNA shuffling (exon shuffling), hybrid enzyme, random-priming in vitro recombination (RPR), stagger extension process (StEP), random-directed mutagenesis in vitro, etc., even with little knowlodge of spatial structure and catalytic mechanism of enzymes in advance. The process that would take millions of years in nature can in principle be accomplished in the test within several years.