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低压段DHPM作用对木瓜蛋白酶结构影响的荧光光谱分析

Fluorescence Spectra Analysis of Papain Structure Treated by Dynamic High Pressure Microfluidization in Low Pressure Ranges

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摘要

以荧光光谱为检测手段研究了低压段(60~100 MPa)动态高压微射流作用(DHPM)对木瓜蛋白酶(Papain)分子结构的影响。 结果显示, 在60~100 MPa下经DHPM处理后, Papain, Tyr残基和Trp残基荧光强度均有不同程度降低, 且随着处理压力的增加, Papain, Tyr残基和Trp残基的荧光强度逐渐增大, Papain, Trp残基荧光发射峰的位置分别从未处理时的334和277.5 nm逐渐红移至100 MPa处理后的335和278.5 nm; 在0~4 ℃放置24 h后, Papain, Tyr残基和Trp残基的荧光光谱均基本保持了0 h时的变化趋势。 表明低压DHPM处理后, Papain中Trp残基逐渐暴露出来, 并形成了较为稳定的新分子构象。

Abstract

The present research investigated the effect of dynamic high pressure microfluidization (DHPM) in low pressure ranges (60-100 MPa) on the molecular structure of papain with the help of fluorescence spectra as the detection method. The result showed that after the treatment of DHPM at 60-100 MPa, the fluorescence intensity of papain, tryptophan (Trp) and tyrosine (Tyr) residues all decreased to different extents. Meanwhile, with the increase in the treatment pressure, their fluorescence intensity would gradually increase and the fluorescence emission peak would gradually red shift from 334 and 277.5 nm before treatment to 335 and 278 nm after 100 MPa treatment for papain and Trp residue respectively; after the treatment, with the samples being placed at 0-4 ℃ for 24 h, the fluorescence spectra of papain, Tyr and Trp residues in various experiment groups basically maintained the same changing tendency compared to that of newly treated samples. Hence, it showed that after the treatment of DHPM in low pressure ranges, the Trp residue of papain was gradually brought to light and formed new and comparatively stable molecular conformation.

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中图分类号:O433.4

基金项目:食品科学与技术国家重点实验室目标导向项目(SKLF-MB-200808)和国家(863计划)重点项目(2007AA100403)资助

收稿日期:2009-02-20

修改稿日期:2009-05-18

网络出版日期:0001-01-01

作者单位    点击查看

刘伟:南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047
谢明勇:南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047
钟业俊:南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047
刘成梅:南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047
官斌:南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047
王倩:南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047

联系人作者:谢明勇(myxie@ncu.edu.cn)

备注:刘伟, 1972年生, 南昌大学生命科学与食品工程学院副教授

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引用该论文

LIU Wei,XIE Ming-yong,ZHONG Ye-jun,LIU Cheng-mei,GUAN Bin,WANG Qian. Fluorescence Spectra Analysis of Papain Structure Treated by Dynamic High Pressure Microfluidization in Low Pressure Ranges[J]. Spectroscopy and Spectral Analysis, 2010, 30(2): 387-390

刘伟,谢明勇,钟业俊,刘成梅,官斌,王倩. 低压段DHPM作用对木瓜蛋白酶结构影响的荧光光谱分析[J]. 光谱学与光谱分析, 2010, 30(2): 387-390

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