中国食品学报2017，Vol.17Issue(1)：148-153,6.DOI:10.16429/j.1009-7848.2017.01.019
苦瓜核糖核酸酶的分离纯化与性质分析
Purification and Characterization of RNase from Momordicacharantia L.
林岱 1吴文峰 2柯李晶 1周建武 1饶平凡1
作者信息
- 1. 浙江工商大学食品与生物工程学院 杭州310018
- 2. 福州大学生物工程研究所 福州350002
- 折叠
摘要
Abstract
Objective:To isolate and purify a ribonuclease from Momordicacharantia L.and study the enzymatic and biological characterizations of the ribonuclease.Methods:Ethanol precipitation,C1s reversed phase high performance liquid chromatography (RP-HPLC),Tricine-SDS-polyacrylamide gel electrophoresis and isoelectric focusing were used for isolation,purication and identification of the ribonuclease.The enzyme activity of ribonuclease was detected using tRNA from yeast as substrate.Results:A ribonuclease,designated MC-RNase,showed a high purity by Tricine-SDS-PAGE and RP-HPLC.Its N-terminal sequence was determined as VNEFDYYQVVLQWQPAT.The molecular weight and isoelectric point,was estimated to be 14 ku and 8.1.Enzymic experiment showed the optimum temperature is 50 ℃,and optimum pH is 5,Pb2+ can increase the enzyme activity,while Zn2+ 、Fe2+ 、Fe3+ and Mg2+ inhibited the enzyme activity of MC-RNase.The result of cytotoxicity test showed MC-RNasehad no effect on the proliferation of L-02 cells and Hep-G2 cells in vitro.Conclusion:Isolation and identification of a novel ribonuclease from fresh Momordicacharantia L.关键词
苦瓜/分离纯化/核糖核酸酶/表征Key words
bitter gourd/purification/ribonuclease/characterization引用本文复制引用
林岱,吴文峰,柯李晶,周建武,饶平凡..苦瓜核糖核酸酶的分离纯化与性质分析[J].中国食品学报,2017,17(1):148-153,6.
