The site-directed mutagenesis of gastrodia anti-fungal protein mannose-binding sites and its expression in Escherichia coli.
Gastrodia anti-fungal protein (GAFP) displays strong inhibitory activity against certain fungal pathogens. Five GAFP analogues with different mutations at mannose-binding sites and the wild-type one were expressed and purified in Escherichia coli. The inhibitory analysis of the purified various GAFPs against the growth of Trichoderma viride indicates that single amino acid mutated-type GAFPs have inhibitory activity, but its activity is much less than the wild-type one. The double and triplicate amino acids mutated GAFPs have very low inhibitory activity. For the first time it was proved that GAFP mannose-binding sites play key role in anti-fungi process.
Wang
Peng
P
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, PR China.
Wang
Yiqin
Y
Sa
Qila
Q
Li
Wenbin
W
Sun
Yongru
Y
D000935:Antifungal Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D015342:DNA Probes; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D031668:Gastrodia / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology; D015870:Gene Expression; D037601:Mannose-Binding Lectin / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry; D014242:Trichoderma / Q000187:drug effects / Q000254:growth & development
D000935:Antifungal Agents; D015342:DNA Probes; D037601:Mannose-Binding Lectin; D011994:Recombinant Proteins