Sugiyama H, Idekoba C, Kajino T, Hoshino F, Asam O, Yamada Y, Udaka S
Toyota Central Research & Development Laboratories Inc., Aichi, Japan.
Biosci Biotechnol Biochem. 1993 Oct;57(10):1704-7. doi: 10.1271/bbb.57.1704.
A protein disulfide isomerase (PDI) was purified to homogeneity from the thermophilic fungus Humicola insolens by a rapid three-step procedure, anion-exchange chromatography, concanavalin A-affinity chromatography, and reverse phase high performance liquid chromatography. Forty-one micrograms of PDI was obtained from 100 g of wet mycelium. Concanavalin A-Sepharose chromatography is available for purification of the fungal PDI, indicating that the enzyme is also glycosylated like the yeast PDI. The fungal PDI exists as a dimer (2 x 60 kDa), has a pI of 3.5, and is fairly heat-stable. The amino acid composition of the PDI is similar to those of yeast and bovine liver PDI, and the high content of acidic amino acid residues agrees with the lower acidic pI.
通过快速三步法，即阴离子交换色谱法、伴刀豆球蛋白A亲和色谱法和反相高效液相色谱法，从嗜热真菌特异腐质霉中纯化得到了一种蛋白质二硫键异构酶（PDI），且达到了同质纯。从100克湿菌丝体中获得了41微克的PDI。伴刀豆球蛋白A - 琼脂糖色谱法可用于纯化该真菌PDI，这表明该酶也像酵母PDI一样进行了糖基化修饰。该真菌PDI以二聚体形式（2×60 kDa）存在，pI为3.5，且具有相当的热稳定性。PDI的氨基酸组成与酵母和牛肝PDI相似，酸性氨基酸残基的高含量与较低的酸性pI相符。
