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Enhancing thermo-induced recombinant protein production in Escherichia coli by temperature oscillations and post-induction nutrient feeding strategies.

Abstract：

:Traditional strategies for production of thermo-induced recombinant protein in Escherichia coli consist of a two-phase culture, with an initial growth stage at low temperature (commonly 30°C) followed by a production stage where temperature is increased stepwise (commonly up to 42°C). A disadvantage of such strategies is that growth is inhibited upon temperature increase, limiting the duration of the production stage and consequently limiting recombinant protein production. In this work, a novel oscillatory thermo-induction strategy, consisting on temperature fluctuations between 37 and 42°C or 30 and 42°C, was tested for improving recombinant protein production. In addition, the induction schemes were combined with one of three different nutrient feeding strategies: two exponential and one linear. Recombinant human preproinsulin (HPPI), produced under control of the λP(L)-cI857 system in the E. coli BL21 strain, was used as the model protein. Compared to the conventional induction scheme at constant temperature (42°C), longer productive times were attained under oscillatory induction, which resulted in a 1.3- to 1.7-fold increase in maximum HPPI concentration. Temperature oscillations led to a 2.3- to 4.0-fold increase in biomass accumulation and a decrease of 48-62% in the concentration of organic acids, compared to conventional induction. Under constant induction, growth ceased upon temperature increase and the maximum concentration of HPPI was 3.9 g/L, regardless of the post-induction feeding strategy used. In comparison, the combination of temperature oscillations and a high nutrient-feeding rate allowed sustained growth after induction and reaching up to 5.8 g/L of HPPI.

journal_name

J Biotechnol

journal_title

Journal of biotechnology

authors

Caspeta L,Lara AR,Pérez NO,Flores N,Bolívar F,Ramírez OT

doi

10.1016/j.jbiotec.2013.06.001

keywords:

["Bacteriophage lambda P(L) promoter","E. coli","Recombinant protein","Temperature induction","Temperature oscillations"]

subject

Has Abstract

pub_date

2013-08-10 00:00:00

pages

47-55

issue

eissn

0168-1656

issn

1873-4863

pii

S0168-1656(13)00249-6

journal_volume

167

pub_type

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Enhancing thermo-induced recombinant protein production in Escherichia coli by temperature oscillations and post-induction nutrient feeding strategies.