源于银杏叶片中一个抗真菌蛋白(GAFP-1)的分离、纯化和鉴定

Isolation,Purification and Identification of An Antifungal Protein from Leaves of Ginkgo biloba L.

经过 8 0 %饱和度硫酸铵沉淀、几丁质亲和层析和SephadexG 75的凝胶过滤层析 3个步骤 ,从银杏(GinkgoBilobaL )的叶片中纯化获得 1个抗真菌蛋白 ,把它命名为GAFP 1。以SDS PAGE胶为基础进行电泳分析 ,GAFP 1的分子量约为 30kD。氨基酸组分分析显示 ,1mlGAFP 1的甘氨酸、谷氨酸和天冬氨酸残基较其它氨基酸残基多。抑菌分析表明 :1μg或 2 μg的GAFP 1能够抑制水稻纹枯病菌和棉花炭疽病菌菌丝的生长 ,但对水稻稻瘟病菌和小麦纹枯病菌菌丝的生长没有抑制效果。GAFP 1的N 端 10个氨基酸残基为Asp Pro Gly Cys Asn Val Leu Glu Asp Ile ,以这 10个氨基酸残基为基础进行同源性比较发现 ,它与蛋白质数据库中已知的蛋白质没有同源性 ,表明GAFP 1可能是

An antifungal protein,designated GAFP 1,was purified from leaves of Ginkgo biloba L.by 80% saturation ammonium sulfate,affinity chromatography on regenerated chitin and gel filtration on Sephadex G 75.GAFP 1 had molecular weight about 30 kD on the basis of SDS PAGE.Amino acid analysis showed that it had higher contents of residues of Gly,Asp and Glu than other amino acids.In in vitro antifungal assays,1 μg or 2 μg of GAFP 1 could heavily inhibit the hyphal growth of Pellicularia sasakii Ito and Fusarium moniliforme ,but it didnt show antifungal activity to Magnaporthe grisea and Rhizoctoniz cerealis .10 amino acids of GAFP 1 in the N terminal were Asp Pro Gly Cys Asn Val Leu Glu Asp Ile.Similarity comparison showed that GAFP 1 had not any similarity with known proteins in protein databases.The results suggested GAFP 1 might be a new antifungal protein. [WT5”HZ]