Kinetics of inclusion body formation studied in intact cells by FT-IR spectroscopy

被引：74
作者：
Ami, D [1 ]
Natalello, A [1 ]
Gatti-Lafranconi, P [1 ]
Lotti, M [1 ]
Doglia, SM [1 ]
机构：
[1] Univ Milan Bicocca, Dipartimento Biotecnol & Biosci, I-20126 Milan, Italy
来源：
FEBS LETTERS | 2005年 / 579卷 / 16期
关键词：
inclusion body; protein aggregation; kinetics of recombinant protein production; FT-IR microspectroscopy; lipase;
D O I：
10.1016/j.febslet.2005.04.085
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The aggregation of a recombinant lipase as inclusion bodies (IBs) was studied directly within intact Escherichia coli cells by FT-IR microspectroscopy. Through this approach, it was possible to monitor in real time the different kinetics of IB formation at 37 and 27 degrees C, in excellent agreement with the results of the SDS-PAGE analysis. Furthermore, insights on the residual native-like structure of the expressed protein within IB - both isolated and inside cells - were obtained by the secondary structure analysis of the Amide I band in the IB FT-IR spectra. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码：3433 / 3436
页数：4
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