CHEN Di,LIU Yin-yan,YANG Jin-zhu,et al.Separating Soybean Trypsin Inhibitors with Solid-Phase Enzyme Method[J].Soybean Science,2010,29(06):1056-1058.[doi:10.11861/j.issn.1000-9841.2010.06.1056]
固相酶法分离大豆胰蛋白酶抑制剂研究
- Title:
- Separating Soybean Trypsin Inhibitors with Solid-Phase Enzyme Method
- 文章编号:
- 1000-9841（2010）06-1056-03
- Keywords:
- Solid-Phase enzyme method; Soybean; Trypsin Iinhibitors
- 分类号:
- TQ646.5
- 文献标志码:
- A
- 摘要:
- 以硅胶-壳聚糖为载体，通过戊二醛偶联胰蛋白酶，制备固定化酶，分离纯化大豆中胰蛋白酶抑制剂。对戊二醛浓度、反应温度、酶浓度等影响固定化酶制备的因素进行了研究。戊二醛的浓度为2.0%，反应温度为20℃，胰蛋白酶与载体比为15∶1 000时，分离纯化的大豆胰蛋白酶抑制剂活性最高，达1872.51 U?mg-1，比粗品高46.22倍。结果表明通过该法能够获得较高活性大豆胰蛋白酶抑制剂，为大豆胰蛋白酶抑制剂的开发利用提供了依据。
- Abstract:
- In order to separate and purify the soybean trypsin inhibitors, immobilized trypsin was prepared by coupling siliconechitosan activated carriers with trypsin using glutaraldehyde as coupling agents. The influence factors of coupling reaction including glutaraldehyde concentration, reaction temperature and trypsin concentration were investigated and the results showed that the best conditions were as follow: the concentration of glutaraldehyde was 2.0%, the temperature was 20℃, and the ratio of trypsin with silicone-chitosan was 15∶1 000. The activity of soybean trypsin inhibitors isolated by the above immobilized trypsin was 1872.51 U?mg-1, and the purified degree increased 46.22 times. The results suggested that soybean trypsin inhibitors with higher activity can be obtained by this method and it is a better way to exploit the soybean trypsin inhibitors.
参考文献/References:
[1]Brik Y. The Bowman-Birk inhibitor. Trypsin and chymotrypsin inhibitor from soybeans[J]. International Journal of Peptide Protein Research 1985, 25(2): 113-131. [2]蔡祖花，王凤山，张天民. 胰蛋白酶抑制剂临床研究概况[J]. 中国生化药物杂志2000, 21（3）：157-159.（Cai Z H, Wang F S, Zang T M. Clinical reseach status of trypsinInhibitors[J]. Chinese Journal of Biochemical Pharmaceutics 2000, 21(3):157-159.） [3]孙晓丰，余蓉，曾蓉等. 固定化胰蛋白酶的制备[J]. 华西药学杂志, 2003, 18(3): 176-178. (Sun X F, Yu R, Zeng R, et al. Preparation of immobilized trypsin[J]. West China Journal of Pharmaceutical Sciences, 2003, 18(3): 176-178.) [4]王英男，修建成，富校轶，等. 豆制品中胰蛋白酶抑制剂活性测定方法研究[J]. 大豆通报, 2006(2): 31-32.(Wang Y N, Xiu J C, Fu X Y，et al. Measurement of trypsin inhibitor activities in soybean product [J]. Soybean Bulletin, 2006(2): 31-32.) [5]宋杨，侯司，赵辉，等. 壳聚糖固定化酶一步纯化抑肽酶研究[J]. 生物学杂志, 2000, 17(1)：18-21.(Song Y, Hou S, Zhao H, et al. A new method of preparation of high-purity aprotininumby chemica1 modefied trypsin of chitosan[J]. Journal of Biology, 2000, 17(1): 18-21.) [6]曹维强，王静，陈鹏.壳聚糖的构效关系及其在食品中的应用[J]食品研究与开发，2006,27（5）：165-168.（Cao W Q，Wang J, Chen P. The relationship of chitosan structure and functional properties and its application food industry[J]. Food research and development， 2006,27(5):165-168.）
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备注/Memo
第一作者简介：陈滴（1957-），男，高级工程师，研究方向为药物化学教学与科研工作。E-mail：chendi@jlu.edu.cn。