Cloning of a full-length cDNA encoding ent-kaurene synthase from Gibberella fujikuroi: functional analysis of a bifunctional diterpene cyclase.
We report here the nucleotide sequence of a full-length cDNA encoding ent-kaurene synthase that was isolated by a reverse-transcription polymerase chain reaction from Gibberella fujikuroi (Gcps/ks). This cDNA encodes 952 amino acid residues with a relative molecular mass of 107 kDa. The sequence similarity between Gcps/ks and ent-kaurene synthase of the gibberellin A1-producing fungus, Phaeosphaeria sp. L487, is very high, suggesting that Gcps/ks is also a bifunctional diterpene cyclase. Its recombinant protein expressed in Escherichia coli converted geranylgeranyl diphosphate to copalyl diphosphate and ent-kaurene.
Toyomasu
T
T
Department of Bioresource Engineering, Yamagata University, Tsuruoka-shi, Japan. toyomasu@tds1.tr.yamagata-u.ac.jp
Kawaide
H
H
Ishizaki
A
A
Shinoda
S
S
Otsuka
M
M
Mitsuhashi
W
W
Sassa
T
T
D019883:Alkyl and Aryl Transferases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005874:Gibberella / Q000201:enzymology*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D019883:Alkyl and Aryl Transferases; C105712:ent-kaurene synthetase B