朱桔Mn-SOD的纯化、鉴定及浓度梯度胶电泳在其中的应用

植物研究 ›› 2004, Vol. 24 ›› Issue (2): 240-244.

朱桔Mn-SOD的纯化、鉴定及浓度梯度胶电泳在其中的应用

程光宇, 吴国荣, 储慧君

南京师范大学生命科学学院, 南京 210097

收稿日期:2003-11-15 出版日期:2004-06-15 发布日期:2016-06-14
作者简介:程光宇(1954-), 男, 高级工程师, 主要从事植物生物活性物质研究及开发工作。
基金资助:
江苏省自然科学基金资助项目(1998SWXOSZ001)

Purification and differentiation of Mn-SOD from Citrus erythrosa and application of gel concentration gradient electrophoresis

CHENG Guang-Yu, WU Guo-Rong, CHU Hui-Jun

School of Life Science, Nanjing No rmal University, Nanjing 210097

Received:2003-11-15 Online:2004-06-15 Published:2016-06-14

摘要/Abstract

摘要： 朱桔叶中存在Mn-SOD、Fe-SOD和CuZn-SOD三种类型,在10%的PAGE中有三条活性带,其中Mn-SOD的Rf值大于Fe-SOD与CuZn-SOD₁的Rf值相同;在4%~35%的梯度胶电泳中有四条活性带,而Mn-SOD的R_f值最小,表明该酶带有较高的电荷密度。Mn-SOD占总活性的20%左右,已被纯化到均一程度。该酶的比活性为1 249 U/mg,分子量和亚基分子量分别为54.0 kD和26.6 kD,在紫外区最大吸收值为280 nm,在95℃处理15 min仍保留了46%的酶活性,等电点为5.06。该酶的活性不被KCN、H₂O₂抑制,但对1% SDS和氯仿-乙醇液敏感。

关键词: 朱桔, Mn-SOD, 纯化, 性质, 电荷密度

Abstract: There were Mn-SOD,Fe-SOD and CuZn-SOD in the crude extract of Citrus erythrosa Tanaka leaves.In a 10% PAGE three active bands were seen,and R fvalue of Mn-SOD was big than that of Fe-SOD and was same as that of CuZn-SOD₁.While in a 4%~35% gel concentration gradient electrophoresis,four active bands were seen and Mn-SOD's R fvalue was the smallest.This means that the molecular weight of Mn-SOD was the biggest and it had a high charge density.Mn-SOD,which occupied about 20% of the total SOD activity,was purified to homogeneity by means of ammonium sulfate precipitation,Sephadex G-100 gel filtration,DEAE-Sepharose and CMsub>52 chromatography etc.Its specific activity was 1 249 U/mg.Its molecular and subunit weight were 54.0 kD and 26.6 kD,respectively.It had one absorption maximum which was at 280 nm.And it was stable to heat treatment,46% of its activity was retained at 95℃ for 15 min.Isoelectric focusing electrophoresis showed that it had a pI of 5.06.This enzyme was insensitive to KCN and H₂O₂,but was sensitive to 1% SDS or mixtures of chloroform and ethanol (3:5 v/v).

Key words: Citrus erythrosa Tanaka, Mn-SOD, purification, characterization, charge density

程光宇, 吴国荣, 储慧君. 朱桔Mn-SOD的纯化、鉴定及浓度梯度胶电泳在其中的应用[J]. 植物研究, 2004, 24(2): 240-244.

CHENG Guang-Yu, WU Guo-Rong, CHU Hui-Jun. Purification and differentiation of Mn-SOD from Citrus erythrosa and application of gel concentration gradient electrophoresis[J]. Bulletin of Botanical Research, 2004, 24(2): 240-244.

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