摘要
苦荞粉经缓冲液提取后,用Trypsin-SESA-Sephadex G-75亲和色谱分离纯化,得到苦荞胰蛋白酶抑制剂,此抑制剂进一步在CM-52,DEAE-52上分离纯化,得到五个组份。对其中三个主要组份(BTI-Ⅰ,Ⅱ,Ⅲ)的研究表明,它们的消光系数(E_(1cm)^(1％) 280nm)分别为4.1,4.8和7.8,最大吸收光谱分别为280nm,274nm和277nm。纯化倍数达160-167倍。它们在酸性条件下均具有很高的耐热性,其中以BTI-Ⅱ的热稳定性最高。用纯化后的BTI-Ⅱ作配基,制成Trypsin inhibitor-SESA-Sephadex G-75亲和吸附剂,可用于纯化胰蛋白酶。
The buckwheat (Fagopyrum tataricum) trypsin inhibitor was abtained by affinity chromatography with trypsin-SESA-Sephadex G—75 and five components of it were purified further by ion exchange chromatography on CM—cellulose—52 and DEAE—cellulose—52. Studies on three main components (BTI—Ⅰ,Ⅱ,Ⅲ) showed that their extinction coefficients (E_(1ck)^(1％) at 280nm) were 4.1, 4.8 and 7.8 and their maximum absorptions were observed at 280nm, 274nm, 277nm respectively. An overall purification of three inhibitors was about 160 fold. These inhibitors, especially BTI—Ⅱ, were highly heat stable under the acid condition. The trypsin could be purified by affinity chromatography with trypsin inhibitor-SESA-Sephadex G—75 which was prepared with BTI-Ⅱ.
关键词
苦荞麦
荞麦
胰蛋白酶
抑制剂
Trypsin inhibitors
Affinity chromatography
Buckwheat
Fagopyrum tataricum.
