The solubility of the ribotoxin α‐sarcin, produced as a recombinant protein in Escherichia coli, is increased in the presence of thioredoxin

• Published in: Letters in applied microbiology Vol. 30; no. 4; pp. 298 - 302
• Main Authors: García‐Ortega, L., Lacadena, J., Lacadena, V., Masip, M., De Antonio, C., Martínez‐Ruiz, A., Martínez del Pozo, Á.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.04.2000; Blackwell Science
• Subjects: a-Sarcin; Aspergillus - genetics; Aspergillus - metabolism; Biological and medical sciences; Biotechnology; Endoribonucleases - chemistry; Endoribonucleases - genetics; Endoribonucleases - metabolism; Escherichia coli; Escherichia coli - genetics; Escherichia coli - growth & development; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Fungal Proteins; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; Modification of gene expression level; Plasmids - genetics; Protein Conformation; Protein Synthesis Inhibitors - chemistry; Protein Synthesis Inhibitors - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; ribotoxins; Solubility; thioredoxin; Thioredoxins - metabolism; Enzyme; Gene coexpression; Escherichia coli; Solubility; Environmental factor; Esterases; Gene expression; Thioredoxin; Fungi; Toxin; rRNA endonuclease; Enzymatic activity; Aspergillus giganteus; Production; Microorganism culture; Bacteria; Hydrolases; Fungi Imperfecti; Recombinant protein; Enterobacteriaceae; Thallophyta
• ISSN: 0266-8254; 1472-765X; 1365-2673
• DOI: 10.1046/j.1472-765x.2000.00714.x

The yield of purified recombinant α‐sarcin increases approximately three‐ to fourfold when this toxin is co‐expressed in Escherichia coli with thioredoxin. This increased production is attributed to the existence, in the presence of thioredoxin, of a reducing environment which allows rearrangement of incorrect disulphide bonds to produce the soluble native conformation. The protein thus produced retains the structural, spectroscopic and enzymatic features of the natural fungal α‐sarcin.