Escherichia coli expression systems

Anakinra is a nonglycosylated form of the human IL-1 receptor antagonist (IL-lra). It is produced in a recombinant Escherichia coli expression system and has an additional methionine residue at its amino terminus. In rheumatoid arthritis patients, the amount of naturally occurring IL-lra in the synovial fluid is not sufhcient to counteract the high levels of locally produced IL-1. Anakinra acts as a competitive antagonist of the type 1 IL-1 receptor and decreases the pain and inflammation produced by IL-1. It is administered as a daily subcutaneous injection. 

Pliickthun, A. (1991) Antibody engineering advances from the use of Escherichia coli expression systems. Bio/Technology 9,545-551. 

Maruyama, N., Ichise, K., Katsube, T., Kishimoto, T., Kawase, S., Matsumura, Y., Takeuchi, Y., Sawada, T., Utsumi, S. 1998. Identification of major wheat allergens by means of Escherichia coli expression system. EurJBiochem 255 739-745. 

In an Escherichia coli expression system for the aqualysin I precursor, the precursor is processed autoproteolytically into the mature 28-kDa enzyme by treatment at 65 ° C.23) In this case, the N-terminal pro-sequence is required for the production of active enzyme and functions to stabilize the precursor structure.283 The C-terminal pro-sequence is not essential for the production of active aqualysin 1,293 but seems to be involved in the translocation of the precursor across the cytoplasmic membrane.303 In a Thermus thermophilus expression system,313 the C-terminal pro-sequence is required for the production and extracellular secretion of active aqualysin I.323 In an E. coli expression system for the subtilisin E gene, the N-terminal pro-sequence is essential for the production of active enzyme,333 as in the case of aqualysin I. The requirement of the pro-sequence is also shown in vitro for the refolding of the inactive mature protein to an active enzyme.34 353 The functions of the N-terminal pro-sequences of aqualysin I and subtilisin E seem to be similar. 

A breakthrough in the study of HK97 was achieved when an Escherichia coli expression system was developed in which gene 4 and gene 5 products alone were shown to assemble and mature in a manner identical with the authentic virus (Xie and Hendrix, 1995). Expansion was triggered by... 

The cDNA and corresponding primary amino acid sequences of several CPRs including rat , rabbit- , and human were obtained by the mid-1980s, and the development of Escherichia coli expression systems paved the way for detailed molecular characterization of the polypeptide through site-directed mutagenesis. The three-dimensional structure of rat CPR was determined by X-ray crystallography in 1997 by Kim and coworkers -, providing the structural prototype for dual flavin oxidoreductases. 

Escherichia coli expression systems where the target proteins are secreted into inclusion bodies and have to be refolded to obtain their full biological activity. 

Li, Z., Kessler, W., van den Henvel, J., Rinas, U. (2011). Simple defined antoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems. Applied Microbiology and Biotechnology, 91, 1203—1213. 

Agematu H, Matsumoto N, Fuji Y, Kabumoto H, Doi S, Machida K, Ishikawa J, Arisawa A (2006) Hydroxylation of testosterone by bacterial cytochromes P450 using the Escherichia coli expression system. Biosci Biotechnol Biochem 70 307-311... 

Kitazume T, Tanaka A, Takaya N, Nakamura A, Matsuyama S, Suzuki T, Shoun H (2002) Kinetic analysis of hydroxylation of saturated fatty acids by recombinant P450foxy produced by an Escherichia coli expression system. Eur J Biochem 269 2075-2082... 

Zelasko S, PalariaA, Das A (2013) Optimizations to achieve high-level expression of cytochrome P450 proteins using Escherichia coli expression systems. Protein Expr Purif20 00143-00145... 

A thermally stable NHase from Comamonas testosteroni 5-MGAM-4D (ATCC 55 744) [22] was recombinantly expressed in Escherichia coli, and the resulting transformant cells immobilized in alginate beads that were subsequently chemically cross-linked with glutaraldehyde and polyethylenimine. This immobilized cell catalyst (at 0.5 % dew per reaction volume) was added to an aqueous reaction mixture containing 32wt% 3-cyanopyridine at 25 °C, and a quantitative conversion to nicotinamide was obtained. The versatility of this catalyst system was further illustrated by a systematic study of substrates, which included... 

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