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

页码：3433 / 3436

页数：4

共 14 条

[1] The cold-active lipase of Pseudomonas fragi -: Heterologous expression, biochemical characterization and molecular modeling [J].

Alquati, C ;

De Gioia, L ;

Santarossa, G ;

Alberghina, L ;

Fantucci, P ;

Lotti, M .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2002, 269 (13) :3321-3328

[2] FT-IR study of heterologous protein expression in recombinant Escherichia coli strains [J].

Ami, D ;

Bonecchi, L ;

Calì, S ;

Orsini, G ;

Tonon, G ;

Doglia, SM .

BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2003, 1624 (1-3) :6-10

[3] Protein aggregation as bacterial inclusion bodies is reversible [J].

Carrió, MM ;

Villaverde, A .

FEBS LETTERS, 2001, 489 (01) :29-33

[4] Fine architecture of bacterial inclusion bodies [J].

Carrió, MM ;

Cubarsi, R ;

Villaverde, A .

FEBS LETTERS, 2000, 471 (01) :7-11

[5] Dynamics of in vivo protein aggregation:: building inclusion bodies in recombinant bacteria [J].

Carrió, MM ;

Corchero, JL ;

Villaverde, A .

FEMS MICROBIOLOGY LETTERS, 1998, 169 (01) :9-15

[6]

DEBERNARDEZ CE, 2001, CURR OPIN BIOTECH, V12, P202

[7]

Haris P I, 1994, Methods Mol Biol, V22, P183

[8] FTIR spectroscopic characterization of protein structure in aqueous and non-aqueous media [J].

Haris, PI ;

Severcan, F .

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 1999, 7 (1-4) :207-221

[9] Secondary structure, conformational stability and glycosylation of a recombinant Candida rugosa lipase studied by Fourier-transform infrared spectroscopy [J].

Natalello, A ;

Ami, D ;

Brocca, S ;

Lotti, M ;

Doglia, SM .

BIOCHEMICAL JOURNAL, 2005, 385 :511-517

[10] NATIVE-LIKE SECONDARY STRUCTURE IN INTERLEUKIN-1-BETA INCLUSION-BODIES BY ATTENUATED TOTAL REFLECTANCE FTIR [J].

OBERG, K ;

CHRUNYK, BA ;

WETZEL, R ;

FINK, AL .

BIOCHEMISTRY, 1994, 33 (09) :2628-2634

← 1 2 →