Cloning and functional expression in Escherichia coli of a cDNA encoding cardenolide 16'-O-glucohydrolase from Digitalis lanata Ehrh.
A clone of cardenolide 16'-O-glucohydrolase cDNA (CGH I) was obtained from Digitalis lanata which encodes a protein of 642 amino acids (calculated molecular mass 73.2 kDa). The amino acid sequence derived from CGH I showed high homology to a widely distributed family of beta-glucohydrolases (glycosyl hydrolases family 1). The recombinant CGH I protein produced in Escherichia coli had CGH I activity. CGH I mRNA was detected in leaves, flowers, stems and fruits of D. lanata.
Framm
J J
JJ
Institut für Pharmazeutische Biologie, Martin-Luther-Universität, Hoher Weg 8, D-06120 Halle (Saale), Germany.
Peterson
A
A
Thoeringer
C
C
Pangert
A
A
Hornung
E
E
Feussner
I
I
Luckner
M
M
Lindemann
P
P
D000595:Amino Acid Sequence; D015152:Blotting, Northern; D002298:Cardenolides / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D004070:Digitalis / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D005959:Glucosidases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins; D010946:Plants, Medicinal; D010947:Plants, Toxic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D002298:Cardenolides; D018076:DNA, Complementary; D010940:Plant Proteins; D012333:RNA, Messenger; C113503:Cardenolide 16-O-glucohydrolase, Digitalis lanata; D005959:Glucosidases