Direct measurements of IPTG enable analysis of the induction behavior of E. coli in high cell density cultures.

Microbial Cell Factories
Alfred Fernández-CastanéJosep López-Santín

Abstract

The E. coli lac operon and its components have been studied for decades, and lac-derived systems are widely used for recombinant protein production. However, lac operon dynamics and induction behavior remain the paradigm of gene regulation. Recently, an HPLC-MS-based method to quantify IPTG in the medium and inside the biomass has been established, and this tool may be useful to uncover the lack of knowledge and allow optimization of biotechnological processes. The results obtained from the study of IPTG distribution profiles in fed-batch, high cell density cultures allowed discrimination between two different depletion patterns of an inducer from the medium to the biomass in E. coli-expressing rhamnulose-1-phosphate aldolase (RhuA). Moreover, we could demonstrate that active transport mediates the uptake of this gratuitous inducer. Additionally, we could study the induction behaviors of this expression system by taking into account the biomass concentration at the induction time. In the bistable range, partial induction occurred, which led to intermediate levels of RhuA activity. There was a direct relationship between the initial inducer concentrations and the initial inducer transport rate together with the specific activity...Continue Reading

References

Oct 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·Y Kao-HuangP H von Hippel
Mar 1, 1996·Trends in Biotechnology·S Y Lee
Dec 5, 1998·Journal of Molecular Biology·J MüllerB Müller-Hill
Aug 3, 2001·Nature Reviews. Molecular Cell Biology·H R KabackA B Weinglass
May 14, 2003·The Journal of Cell Biology·José M G VilarStanislas Leibler
Feb 1, 1959·Biochimica Et Biophysica Acta·L A HERZENBERG
Feb 20, 2004·Nature·Ertugrul M OzbudakAlexander Van Oudenaarden
Mar 2, 2005·Biotechnology and Bioengineering·Anna Maria SandénGen Larsson
Jun 14, 2005·Comptes rendus biologies·Mitchell Lewis
Jan 25, 2006·Nucleic Acids Research·Stefan OehlerBenno Müller-Hill
Jul 15, 1957·Proceedings of the National Academy of Sciences of the United States of America·A Novick, M Weiner
May 11, 2007·Journal of Theoretical Biology·Atul Narang
Aug 21, 2008·Nature Protocols·Bryson D BennettJoshua D Rabinowitz
Feb 27, 2009·Microbial Cell Factories·Frank DelvignePhilippe Thonart
Oct 26, 2010·Carbohydrate Research·Mari Ishikawa, Satoshi Koizumi
Jul 2, 2011·Microbial Cell Factories·Federica VolontèLoredano Pollegioni
Oct 26, 2011·Microbial Cell Factories·Willem M de Vos
Dec 29, 2011·Journal of Biotechnology·Alfred Fernández-CastanéJosep López-Santín

❮ Previous
Next ❯

Citations

Feb 4, 2014·Journal of Biological Engineering·Nicolo' PolitiPaolo Magni
Oct 7, 2015·Bioprocess and Biosystems Engineering·Sushma ChityalaReddy Shetty Prakasham
Aug 1, 2015·Molecular BioSystems·Huy TranAndre S Ribeiro
Nov 5, 2016·Preparative Biochemistry & Biotechnology·Pouria Gholami TilkoHamid Moghimi
Jan 23, 2020·International Journal of Molecular Sciences·Luciana GomesFilipe Mergulhão
Sep 19, 2020·Applied Microbiology and Biotechnology·Balaram MishraMukesh Kumar Gupta

❮ Previous
Next ❯

Software Mentioned

Lab Solutions

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.