蜂王浆中主蛋白成分(MRJP2)的分离纯化及圆二色谱分析

doi:10.7506/spkx1002-6630-201323013

食品科学 ›› 2013, Vol. 34 ›› Issue (23): 57-61.doi: 10.7506/spkx1002-6630-201323013

蜂王浆中主蛋白成分(MRJP2)的分离纯化及圆二色谱分析

胡熠凡1,2，陈忠周2，田文礼1,2，赵亚周1，彭文君1,*

1.中国农业科学院蜜蜂研究所，北京 100093；2.中国农业大学生物学院，农业生物技术国家重点实验室，北京 100193

收稿日期:2012-11-26 修回日期:2013-10-29 出版日期:2013-12-15 发布日期:2014-01-03
通讯作者: 彭文君 E-mail:pengwenjun@vip.sina.com
基金资助:
国家现代农业(蜜蜂)产业技术体系建设专项(NYCYTI-43-KXJ17)；中央级公益性科研院所基本科研业务费专项(2012ZL077)

Circular Dichroism Analysis of MRJP2 Purified from Royal Jelly

HU Yi-fan1,2，CHEN Zhong-zhou2，TIAN Wen-li1,2，ZHAO Ya-zhou1，PENG Wen-jun1,*

1. Institute of Apiculture, Chinese Academy of Agricultural Sciences, Beijing 100093, China；
2. State Key Laboratory of Agro-biotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China

Received:2012-11-26 Revised:2013-10-29 Online:2013-12-15 Published:2014-01-03
Contact: PENG Wen-jun E-mail:pengwenjun@vip.sina.com

摘要/Abstract

摘要：

利用亲和层析、分子筛层析和离子交换层析从新鲜蜂王浆中提取蜂王浆中主蛋白成分(MRJP2)的单一组分，经SDS电泳和质谱鉴定为MRJP2后，再通过圆二色谱技术对MRJP2在不同温度、不同pH值、不同离子浓度下二级结构的变化情况进行检测。结果表明：10～90℃升温过程中，α螺旋比例增加、β折叠和无规则卷曲比例减少，β转角比例变化不大；pH值由4.0上升到9.0过程中，α螺旋比例先减少后增加，β折叠比例在pH5.0时最高，β转角比例基本不变，无规则卷曲比例在pH8.0时最高；蛋白溶液中添加不同金属离子后，Ca2+可以促进α螺旋比例增加，而Mg2+和Zn2+会明显降低α螺旋比例。

关键词: 蜂王浆, MRJP2, 纯化, 二级结构, 圆二色谱

Abstract:

MRJP2, a member of the major royal jelly protein (MRJP) family, was extracted from fresh royal jelly, purified
by affinity chromatography, molecular sieve chromatography and ion exchange chromatography and identified by SDSPAGE
and mass spectrum. The MRJP protein was analyzed by synchrotron radiation circular dichroism (SRCD) to clarify
its secondary structure as a function of temperature, pH and ion concentrations. The results of this study showed that heat treatment
at elevated temperatures from 10 to 90 ℃ resulted in increased proportion of α-helix and decreased proportion of β-strand
and random coil, while the proportion of β-turn kept stable. As the pH varied from 4.0 to 9.0, the proportion of α-helix revealed
an initial decrease followed by an increase, the highest proportions of β-strand and random coil were found at pH 5.0 and 8.0,
respectively, and the proportion of β-turn remained basically unchanged. After different metal ions were added in the protein
solution, the proportion of α-helix was increased by Ca2+ but decreased by Mg2+ and Zn2+.

Key words: royal jelly, MRJP2, purification, secondary structure, circular dichroism (CD)

中图分类号:

TS201.2

胡熠凡1,2，陈忠周2，田文礼1,2，赵亚周1，彭文君1,*. 蜂王浆中主蛋白成分(MRJP2)的分离纯化及圆二色谱分析[J]. 食品科学, 2013, 34(23): 57-61.

HU Yi-fan1,2，CHEN Zhong-zhou2，TIAN Wen-li1,2，ZHAO Ya-zhou1，PENG Wen-jun1,*. Circular Dichroism Analysis of MRJP2 Purified from Royal Jelly[J]. FOOD SCIENCE, 2013, 34(23): 57-61.

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蜂王浆中主蛋白成分(MRJP2)的分离纯化及圆二色谱分析

Circular Dichroism Analysis of MRJP2 Purified from Royal Jelly

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