A novel recombinant system for functional expression of myonecrotic snake phospholipase A(2) in Escherichia coli using a new fusion affinity tag

Protein Expression and Purification
Minae SetoMotonori Ohno

Abstract

A novel recombinant expression system in Escherichia coli was developed using conger eel galectin, namely, congerin II, as an affinity tag. This system was applied for the functional expression of myotoxic lysine-49-phospholipase A(2) ([Lys(49)]PLA(2)), termed BPII and obtained from Protobothrops flavoviridis (Pf) venom. Recombinant Pf BPII fused with a congerin tag has been successfully expressed as a soluble fraction and showed better quantitative yield when folded correctly. The solubility of the recombinant congerin II-tagged BPII increased up to >90% in E. coli strain JM109 when coexpressed with the molecular chaperones GroEL, GroES, and trigger factor (Tf). The tag protein was cleaved by digestion with restriction protease, such as alpha-thrombin or Microbacterium liquefaciens protease (MLP), to obtain completely active recombinant BPII. Thus, the congerin-tagged fusion systems containing the cleavage recognition site for alpha-thrombin or MLP were demonstrated to be highly efficient and useful for producing proteins of desired solubility and activity.

References

Sep 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·T OgawaM Ohno
Apr 22, 1992·Biochimica Et Biophysica Acta·K Muramoto, H Kamiya
Jan 1, 1994·Annual Review of Biophysics and Biomolecular Structure·S J Landry, L M Gierasch
Nov 15, 1993·The Biochemical Journal·A Sivaprasadarao, J B Findlay
Jan 17, 1997·The Journal of Biological Chemistry·O KandrorA L Goldberg
Aug 5, 1998·Toxicon : Official Journal of the International Society on Toxinology·H PanX F Wu
Oct 29, 1998·Toxicon : Official Journal of the International Society on Toxinology·I H Tsai, Y M Wang
Jul 30, 1999·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·K MuramotoH Kamiya
Mar 4, 2000·Applied and Environmental Microbiology·K NishiharaT Yura
Aug 2, 2001·Toxicon : Official Journal of the International Society on Toxinology·C D GiulianiH S Selistre-de-Araujo
Nov 28, 2001·Protein Expression and Purification·D SareenR M Vohra
May 10, 2002·Bioscience, Biotechnology, and Biochemistry·Tomohisa OgawaKoji Muramoto
Apr 1, 1958·Archives of Biochemistry and Biophysics·G L ELLMAN
Mar 17, 2004·Toxicon : Official Journal of the International Society on Toxinology·Lucimara Chioato, Richard J Ward
May 26, 2005·Bioscience, Biotechnology, and Biochemistry·Yoshitaka Kanayama, Yasuo Sakai
Jun 1, 2005·Trends in Biotechnology·David S Waugh
Jan 24, 2006·Protein Expression and Purification·José ArnauJohn Pedersen

❮ Previous
Next ❯

Citations

Jul 20, 2012·The Journal of Biological Chemistry·Mizuki WatanabeTomohisa Ogawa
May 11, 2012·Journal of Pharmaceutical and Biomedical Analysis·Lucas B SilveiraAndreimar M Soares
Oct 25, 2011·Protein Expression and Purification·Daiki TakemoriYugo Iwasaki
Jun 26, 2015·Journal of Bioscience and Bioengineering·Hui WangGuiyang Shi

❮ 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.