FITC标记脂肪酶的稳定性和荧光光谱  

Stability and Fluorescence Spectra of Lipase Labeled with FITC

作　　者：张驰[1] 宋锡瑾[1] 王杰[2] 闻静[3] 

机构地区：[1]浙江大学化学工程与生物工程系,杭州310027 [2]浙江大学化学系,杭州310027 [3]浙江大学a化学工程与生物工程系,杭州310027

出　　处：《应用化学》2008年第12期1381-1384,共4页Chinese Journal of Applied Chemistry

基　　金：国家自然科学基金(20276067)资助项目

摘　　要：以FITC(异硫氰基荧光素)为荧光探针,研究了标记后的脂肪酶的活性、构象、荧光光谱的变化及三者之间的联系。在不同变性条件下,比较了标记与未标记的脂肪酶活性变化及相对应的荧光光谱。结果表明,荧光标记的脂肪酶耐热性能比未标记的酶有所提高,70℃时完全失活。荧光标记的脂肪酶耐酸碱性能增强。在盐酸胍变性剂的条件下,荧光标记的脂肪酶活性下降趋势渐缓,荧光光谱能较好的反映脂肪酶的活性变化。FITC （fluorescein isothiocyanate） that can be linked to lipase as a fluorescence probe was used to study the relationships among the activity of lipase, conformation, and fluorescence spectrum. The changes of activity and fluorescence spectrum of the labeled and unlabeled lipase were examined under various temperatures, pH and different concentrations of guanidine hydrochloride. The results demonstrated that compared with the unlabeled lipase, the thermal stability of the labeled lipase was improved, and it was completely inactive at 70 ℃. The stability of the labelled lipase at different pH was also increased. The activity of the labeled lipase would decrease under different concentrations of guanidine hydroehloride, and fluorescence spectrum of the labeled lipase could reflect the change of the activity.

关 键 词：脂肪酶 异硫氰基荧光素(FITC) 荧光光谱 稳定性 

分 类 号：O641[理学—物理化学]