Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β

Denèfle, Patrice; S, Kovarik; Ciora, Thierry; Gosselet, N; Bénichou, Jean-Claude; Latta, Martine; Guinet, Françoise; Ryter, A; Mayaux, Jean‐François

doi:10.1016/0378-1119(89)90444-7

Cited by 32 publications

(16 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The hIL-2 was not found in the PS fraction, probably because it could form periplasmic inclusion bodies associated to cellmembrane (Betton and Hofnung 1996). Similar to the results obtained here, Denèfle et al (1989) found that hIL-1b transported to periplasm using OmpA and PhoA signal sequences, led the formation of periplasmic inclusion bodies associated to cell-membrane. The post-induction temperature had not effect in the processing of mSPpac-hIL-2 (data non-shown).…”

Section: Cell Fractionation Analysissupporting

confidence: 92%

Modified penicillin acylase signal peptide allows the periplasmic production of soluble human interferon-γ but not of soluble human interleukin-2 by the Tat pathway in Escherichia coli

et al. 2007

View full text Add to dashboard Cite

Production of periplasmic human interferon-gamma (hINF-gamma) and human interleukin-2 (hIL-2) by the Tat translocation pathway in Escherichia coli BL21-SI was evaluated. The expression was obtained using the pEMR vector which contains the Tat-dependent modified penicillin acylase signal peptide (mSPpac) driven by the T7 promoter. The mSPpac-hINF-gamma was processed and the protein was transported to periplasm. Up to 30.1% of hINF-gamma was found in the periplasmic soluble fraction, whereas only 15% of the mSPpac-hIL-2 was processed, but hIL-2 was not found in the periplasmic soluble fraction.

show abstract

Section: Cell Fractionation Analysissupporting

confidence: 92%

Modified penicillin acylase signal peptide allows the periplasmic production of soluble human interferon-γ but not of soluble human interleukin-2 by the Tat pathway in Escherichia coli

et al. 2007

View full text Add to dashboard Cite

show abstract

“…The OmpA delivery system has been successfully demonstrated in a number of studies, e.g., studies using streptokinase (18) and TolAII (35). In addition, incomplete cleavage of the PhoA leader peptide was used to produce human interleukin-1␤ (9,11). This can affect the acquisition of the mature and active form of the heterologous protein.…”

Section: Discussionmentioning

confidence: 99%

Secretion of Active Recombinant Human Tissue Plasminogen Activator Derivatives in Escherichia coli

Manosroi¹,

Tayapiwatana

Götz

et al. 2001

Appl Environ Microbiol

View full text Add to dashboard Cite

The DNA fragment coding for kringle 2 plus serine protease domains (K2S) of tissue plasminogen activator (tPA) was inserted into a phagemid vector, pComb3HSS. In the recombinant vector, pComb3H-K2S, the K2S gene was fused to gpIII of ⌽M13 and linked to the OmpA signal sequence. The resulting gene, rK2S-gpIII, was inducibly expressed in Escherichia coli XL-1 Blue. The protein was presented on the phage particle. To stop the expression of gpIII, a stop codon between K2S and the gpIII gene was inserted by site-directed mutagenesis. This mutated vector, MpComb3H-K2S, was transformed in XL-1 Blue. After induction with IPTG (isopropyl-␤-Dthiogalactopyranoside), rK2S was found both in the periplasm as an inactive form of approximately 32% and in the culture supernatant as an active form of approximately 68%. The secreted form of rK2S was partially purified by ammonium sulfate (55%) precipitation. The periplasmic form was isolated from whole cells by chloroform extraction. The fibrin binding site of kringle 2 was demonstrated in all expressed versions (phage-bound, periplasmic, and secreted forms) using the monoclonal anti-kringle 2 antibody (16/B). Only the secreted form of rK2S revealed a fibrinogen-dependent amidolytic activity with the specific activity of 236 IU/g. No amidolytic activity of rK2S was observed in either the periplasmic or the phage-bound form. The secretion of rK2S as an active enzyme offers a novel approach for the production of the active-domain deletion mutant tPA, rK2S, without any requirements for bacterial compartment preparation and in vitro refolding processes. This finding is an important technological advance in the development of large-scale, bacteriumbased tPA production systems.

show abstract

“…We provide further evidence that the primary sequence of the signal peptide must also be compatible with the mature polypeptide to which it is linked [20,21]. This observation may be explained by thermodynamic considerations of the conformation of the precursors, a matter which really concerns the whole protein and not the signal peptide only [20].…”

Section: Discussionmentioning

confidence: 90%

Expression of the human UDP‐glucuronosyltransferase UGT1¹6 in Escherichia coli

et al. 1994

View full text Add to dashboard Cite

The membrane-bound human liver UDP-glucuronosyltransferase UGTl*6 was expressed in Escherichia colt'. Exchange of the natural signal peptide by the bacterial signal peptides of pelB or OmpT proteins considerably increased the level of expression and, as the natural signal peptide, targeted the protein to the membranes. The extent of maturation of "pelB-UGTl*6 precursor was about 30%. No processing of 'OmpT-UGTl*6 occurred but the processing rate of this precursor could be significantly increased by mutagenesis of the first two amino acid residues of the mature sequence. These expression vectors allowed us to produce high levels of recombinant mature UGTl*6 required for further structural studies.

show abstract

Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β

Cited by 32 publications

References 33 publications

Modified penicillin acylase signal peptide allows the periplasmic production of soluble human interferon-γ but not of soluble human interleukin-2 by the Tat pathway in Escherichia coli

Modified penicillin acylase signal peptide allows the periplasmic production of soluble human interferon-γ but not of soluble human interleukin-2 by the Tat pathway in Escherichia coli

Secretion of Active Recombinant Human Tissue Plasminogen Activator Derivatives in Escherichia coli

Expression of the human UDP‐glucuronosyltransferase UGT1¹6 in Escherichia coli

Contact Info

Product

Resources

About

Heterologous protein export in Escherichia coli: influence of bacterial signal peptides on the export of human interleukin 1β

Cited by 32 publications

References 33 publications

Modified penicillin acylase signal peptide allows the periplasmic production of soluble human interferon-γ but not of soluble human interleukin-2 by the Tat pathway in Escherichia coli

Modified penicillin acylase signal peptide allows the periplasmic production of soluble human interferon-γ but not of soluble human interleukin-2 by the Tat pathway in Escherichia coli

Secretion of Active Recombinant Human Tissue Plasminogen Activator Derivatives in Escherichia coli

Expression of the human UDP‐glucuronosyltransferase UGT116 in Escherichia coli

Contact Info

Product

Resources

About

Expression of the human UDP‐glucuronosyltransferase UGT1¹6 in Escherichia coli