Abstract:
:Iterative saturation mutagenesis (ISM) is a new and efficient method for the directed evolution of functional enzymes. It reduces the necessary molecular biological work and the screening effort drastically. It is based on a Cartesian view of the protein structure, performing iterative cycles of saturation mutagenesis at rationally chosen sites in an enzyme, a given site being composed of one, two or three amino acid positions. The basis for choosing these sites depends on the nature of the catalytic property to be improved, e.g., enantioselectivity, substrate acceptance or thermostability. In the case of thermostability, sites showing highest B-factors (available from X-ray data) are chosen. The pronounced increase in thermostability of the lipase from Bacillus subtilis (Lip A) as a result of applying ISM is illustrated here.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Reetz MT,Carballeira JDdoi
10.1038/nprot.2007.72subject
Has Abstractpub_date
2007-01-01 00:00:00pages
891-903issue
4eissn
1754-2189issn
1750-2799pii
nprot.2007.72journal_volume
2pub_type
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