Open AccessTowards novel biocatalysts via protein design: the case of lipases
Author(s) -
Schmid R.D.,
Menge U.,
Schomburg D.,
Spener F.
Publication year - 1995
Publication title -
fems microbiology reviews
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.91
H-Index - 212
eISSN - 1574-6976
pISSN - 0168-6445
DOI - 10.1111/j.1574-6976.1995.tb00172.x
Subject(s) - lipase , biology , mutagenesis , protein engineering , escherichia coli , rhizopus oryzae , biochemistry , rational design , enzyme , active site , directed evolution , transformation (genetics) , protein design , triacylglycerol lipase , gene , protein structure , genetics , mutation , mutant , fermentation
During the past 3 years, the tertiary structures of several lipases have been solved by X‐ray analysis. The structures revealed unique features such as hydrophobic ‘patches’ on the surface, presumably involved in lipid supersubstrate binding, and a lid structure which covers the active site in the absence of substrate. Only very recently the first X‐ray structure of a bacterial lipase has been solved, and further structural features different from lipases of eukaryotic origin became apparent. Many lipase genes have been cloned and sequenced recently, and expression systems for the preparation of recombinant enzymes in good yields are available. As an example, the lipase from Rhizopus oryzae has been successfully expressed by us in Escherichia coli , and the resulting inclusion bodies were renatured in high yields. Consequently, the mechanism of action of lipases is now being studied via site‐directed mutagenesis, and the rational design of lipases for the selective transformation of substrates is presently addressed in several laboratories.