Directed evolution to generate cycloartenol synthase mutants that produce lanosterol.
Cycloartenol synthase converts oxidosqualene to cycloartenol, a pentacyclic isomer of the animal and fungal sterol precursor lanosterol. We used directed evolution to find cycloartenol synthase residues that affect cyclopropyl ring formation, selecting randomly generated cycloartenol synthase mutants for their ability to genetically complement a yeast strain lacking lanosterol synthase. To increase the likelihood of finding novel mutations, the little-studied Dictyostelium discoideum cycloartenol synthase was used for the mutagenesis. Several catalytically important residues were identified. [reaction: see text]
Meyer
Michelle M
MM
Department of Chemistry, Rice University, 6100 South Main Street, Houston, Texas 77005, USA.
Xu
Ran
R
Matsuda
Seiichi P T
SP
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D019020:Directed Molecular Evolution; D019751:Intramolecular Transferases / Q000235:genetics* / Q000378:metabolism*; D007810:Lanosterol / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics
D007810:Lanosterol; D019751:Intramolecular Transferases; C021923:cycloartenol synthase