Proteome-based identification of fusion partner for high-level extracellular production of recombinant proteins in Escherichia coli

Biotechnology and Bioengineering
Zhi-Gang QianSang Yup Lee

Abstract

Extracellular production of recombinant proteins in Escherichia coli has several advantages over cytoplasmic or periplasmic production. However, nonpathogenic laboratory strains of E. coli generally excrete only trace amounts of proteins into the culture medium under normal growth conditions. Here we report a systematic proteome-based approach for developing a system for high-level extracellular production of recombinant proteins in E. coli. First, we analyzed the extracellular proteome of an E. coli B strain, BL21(DE3), to identify naturally excreted proteins, assuming that these proteins may serve as potential fusion partners for the production of recombinant proteins in the medium. Next, overexpression and excretion studies were performed for the 20 selected fusion partners with molecular weights below 40 kDa. Twelve of them were found to allow fused proteins to excrete into the medium at considerable levels. The most efficient excreting fusion partner, OsmY, was used as a carrier protein to excrete heterologous proteins into the medium. E. coli alkaline phosphatase, Bacillus subtilis alpha-amylase, and human leptin used as model proteins could all be excreted into the medium at concentrations ranging from 5 to 64 mg/L durin...Continue Reading

References

Apr 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·Y HirotaS Yasuda
Dec 1, 1995·Protein Expression and Purification·S W AltmannR A Kastelein
Mar 1, 1996·Trends in Biotechnology·S Y Lee
Jun 6, 1998·Cellular and Molecular Life Sciences : CMLS·A P Pugsley, O Francetic
Oct 3, 1999·Applied and Environmental Microbiology·K OhdanS Okada
Jan 6, 2000·Biotechnology and Bioengineering·K J Jeong, S Y Lee
Sep 13, 2002·Applied Microbiology and Biotechnology·G MikschE Flaschel
Mar 29, 2003·Applied Microbiology and Biotechnology·A ShokriG Larsson
Feb 18, 2004·Applied Microbiology and Biotechnology·J H Choi, S Y Lee
Mar 15, 2005·Biotechnology Advances·F J M MergulhãoG A Monteiro
Apr 5, 2005·Nature Biotechnology·Katariina MajanderBenita Westerlund-Wikström
May 6, 2006·Journal of Proteome Research·M P NandakumarMark R Marten
Jun 8, 2006·Proteomics·Jeong Wook LeeJong-Shin Yoo
Jan 16, 2007·The Journal of Biological Chemistry·Danielle Tullman-ErcekGeorge Georgiou

❮ Previous
Next ❯

Citations

Jun 25, 2010·Applied Microbiology and Biotechnology·Xiang-Yang Fu
Mar 27, 2013·Applied Microbiology and Biotechnology·Kheng Oon LowRosli Md Illias
Jan 19, 2012·Journal of Industrial Microbiology & Biotechnology·Chung-Jr HuangXiaoming Yang
Mar 13, 2013·Journal of Industrial Microbiology & Biotechnology·Iván Muñoz-Gutiérrez, Alfredo Martinez
Jul 15, 2009·Biotechnology Letters·Ye Ni, Rachel Chen
Nov 30, 2011·Proceedings of the National Academy of Sciences of the United States of America·Gregory BokinskyJay D Keasling
Jan 17, 2014·Applied Microbiology and Biotechnology·Shuchi H DesaiShota Atsumi
Oct 25, 2011·Protein Expression and Purification·Daiki TakemoriYugo Iwasaki
Oct 16, 2015·Proteomics·Malvina PapanastasiouAnastassios Economou
Dec 7, 2010·Protein Expression and Purification·Erik VernetMichael Sundström
Jan 5, 2010·Journal of Biotechnology·Mee-Jung HanWon Hee Han
Aug 4, 2015·International Journal of Biological Macromolecules·Hena DharArvind Gulati
May 25, 2015·Journal of Molecular Biology·Christopher W LennonJames C A Bardwell
Feb 11, 2011·Protein Science : a Publication of the Protein Society·Alexander KotzschMichael Sundström
Jun 14, 2015·Biotechnology Advances·Tsutomu Tanaka, Akihiko Kondo
Apr 22, 2014·Bioresource Technology·Vinuselvi ParisuthamSung Kuk Lee
Dec 4, 2012·Protein Expression and Purification·Shefali GuptaSyed Shams Yazdani
Apr 19, 2015·Microbial Cell Factories·Shuchi H DesaiShota Atsumi
Apr 4, 2009·FEMS Yeast Research·Alexandra GrafDiethard Mattanovich
Oct 28, 2016·Biotechnology and Bioengineering·Dongfang GaoQingsheng Qi
Aug 29, 2018·Protein Science : a Publication of the Protein Society·Garry W BuchkoChristopher D Bahl
Aug 2, 2019·Molecular Microbiology·Zhen YanJames C A Bardwell
Aug 9, 2018·Toxins·Victor DavidRussolina Benedeta Zingali
Dec 24, 2018·Microbial Cell Factories·Lisa Ann BurdetteDanielle Tullman-Ercek
Aug 24, 2017·International Journal of Biological Macromolecules·Jashandeep KaurJagdeep Kaur
May 26, 2016·EcoSal Plus·Kyeong Rok ChoiSang Yup Lee

❮ Previous
Next ❯

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.