Directed evolution of enzymes for biocatalysis and the life sciences

被引:42
作者
Williams, GJ
Nelson, AS
Berry, A [1 ]
机构
[1] Univ Leeds, Sch Biochem & Microbiol, Leeds LS2 9JT, W Yorkshire, England
[2] Univ Leeds, Sch Chem, Leeds LS2 9JT, W Yorkshire, England
[3] Univ Leeds, Astbury Ctr Struct Mol Biol, Leeds LS2 9JT, W Yorkshire, England
关键词
protein engineering; directed evolution; enzymes; enzyme activity; DNA shuffling; error-prone PCR;
D O I
10.1007/s00018-004-4234-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Engineering the specificity and properties of enzymes and proteins within rapid time frames has become feasible with the advent of directed evolution. In the absence of detailed structural and mechanistic information, new functions can be engineered by introducing and recombining mutations, followed by subsequent testing of each variant for the desired new function. A range of methods are available for mutagenesis, and these can be used to introduce mutations at single sites, targeted regions within a gene or randomly throughout the entire gene. In addition, a number of different methods are available to allow recombination of point mutations or blocks of sequence space with little or no homology. Currently, enzyme engineers are still learning which combinations of selection methods and techniques for mutagenesis and DNA recombination are most efficient. Moreover, deciding where to introduce mutations or where to allow recombination is actively being investigated by combining experimental and computational methods. These techniques are already being successfully used for the creation of novel proteins for biocatalysis and the life sciences.
引用
收藏
页码:3034 / 3046
页数:13
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