期刊论文

Purification and Characterization of a Ribonuclease from Leucopaxillus tricolor Fruit Bodies

中文

食用菌学报

1005-9873

2019

26

2

59-64

10.16488/j.cnki.1005-9873.2019.02.009

山西农业大学博士科研启动项目(2017YJ33) 山西省优秀博士来晋工作奖励科研项目(SXYBKY201722) 2014年度山西省煤基重点科技攻关项目(FT2014-03).

本校为其他机构

依次采用离子交换层析(DEAE-Cellulose、CM-Cellulose和SP-Sepharose)和凝胶过滤层析(Superdex 75 HR10/300 GL)从三色雷蘑(Leucopaxillus tricolor)子实体中分离纯化得到一种核糖核酸酶(Ribonuclease,RNase)。该酶比活为1406 U/mg,纯化倍数为154,通过HPLC-LTQ-Orbitrap测序得到4条肽段序列,分别为DMAVSYLSR、IGLSSIPR、ILGRFVVDTYK和SGKANAATPK。该核糖核酸酶的相对分子质量为15000,最适反应pH为9.0,最适反应温度为40℃,热稳定性高,80℃孵育1 h后能保持90%以上的活性:Cu~(2+)、Ca~(2+)、Mn~(2+)、Cd~(2+)和Al~(3+)在1.25~10 mmol/L浓度下对该酶活性具有抑制作用,且抑制作用随着金属离子浓度的升...

A ribonuclease (RNase) was purified from Leucopaxillus tricolor fruit bodies by ion exchange chromatography (DEAE-Cellulose,CM-Cellulose and SP-Sepharose) and gel filtration (Superdex 75 HR10/300 GL). The purification fold of this RNase was 154 and the specific activity was 1406 U/mg. Four amino acid sequences were obtained (DMAVSYLSR,IGLSSIPR,ILGRFVVDTYK and SGKANAATPK) by HPLC-LTQ-Orbitrap identification of the RNase. The relative molecular mass of the RNase was estimated to be 15000. The enzyme displayed pH and temperature optima of 9.0 and ...