Design and implementation of a high yield production system for recombinant expression of peptides.

Rodríguez V, Asenjo JA, Andrews BA

Microbial cell factories7 May 2014Cited by 28 (Europe PMC)

PMID24885242PMCIDPMC4022411DOI10.1186/1475-2859-13-65PepBankPUB-24885242

Summary

BackgroundMaking peptide pharmaceuticals involves challenging processes where many barriers, which include production and manufacture, need to be overcome. A non common but interesting research area is related to peptides with intracellular targets, which opens up new possibilities, allowing the modulation of processes occurring within the cell or interference with signaling pathways. However, if the bioactive sequence requires fusion to a carrier peptide to allow access into the cell, the resulting peptide could be such a length that traditional production could be difficult. The goal of the present study was the development of a flexible recombinant expression and purification system for peptides, as a contribution to the discovery and development of these potentially new drugs.ResultsIn this work, a high throughput recombinant expression and purification system for production of cell penetrating peptides in Escherichia coli has been designed and implemented. The system designed produces target peptides in an insoluble form by fusion to a hexahistidine tagged ketosteroid isomerase which is then separated by a highly efficient thrombin cleavage reaction procedure. The expression system was tested on the anticancer peptides p53pAnt and PNC27. These peptides comprise the C-terminal region and the N-terminal region of the protein p53, respectively, fused by its carboxyl terminal extreme to the cell penetrating peptide Penetratin. High yields of purified recombinant fused peptides were obtained in both cases; nevertheless, thrombin cleavage reaction was successful only for p53pAnt peptide release. The features of the system, together with the procedure developed, allow achievement of high production yields of over 30 mg of highly pure p53pAnt peptide per g of dry cell mass. It is proposed that the system could be used for production of other peptides at a similar yield.ConclusionsThis study provides a system suitable for recombinant production of peptides for scientific research, including biological assays.

CC BYText from Europe PMC. Highlighted sentences are the source of results.

Results from this study2

No result from this study comes from a test of the peptide.

Studies that did not test the peptide alone2

In this study, PNC-27 was a lab tool or a carrier, part of a mix, or the subject of a lab method. These results are not evidence about taking PNC-27.

01LabNot a test of PNC-27 aloneIn vitro (cell-free)Other
In recombinant production tests, the PNC27 fusion peptide was obtained at high yield.
In plain wordsIn lab tests, the system produced a high yield of the PNC27 fusion peptide.
Peptide
PNC-27
Outcome
Not applicablePurified recombinant fusion yield
Study
n: not reported in source
Outcome group
Synthesis and manufacturing · Pharmacology
Source sentences
Summary
High yields of purified recombinant fused peptides were obtained in both cases; nevertheless, thrombin cleavage reaction was successful only for p53pAnt peptide release.
2 context sentences(design, population, supporting numbers)
Summary · context
In this work, a high throughput recombinant expression and purification system for production of cell penetrating peptides in Escherichia coli has been designed and implemented.
Summary · context
The expression system was tested on the anticancer peptides p53pAnt and PNC27.
In this study, PNC-27 was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking PNC-27.
Show in the study text Report a problemFND-2488524201
02LabNot a test of PNC-27 aloneIn vitro (cell-free)Other
In recombinant production tests, thrombin cleavage did not successfully release PNC27, although it released p53pAnt.
In plain wordsIn lab tests, thrombin cleavage released p53pAnt but did not successfully release PNC27.
Peptide
PNC-27
Outcome
Not applicableThrombin cleavage release
Study
n: not reported in source
Outcome group
Synthesis and manufacturing · Pharmacology
Source sentences
Summary
High yields of purified recombinant fused peptides were obtained in both cases; nevertheless, thrombin cleavage reaction was successful only for p53pAnt peptide release.
2 context sentences(design, population, supporting numbers)
Summary · context
In this work, a high throughput recombinant expression and purification system for production of cell penetrating peptides in Escherichia coli has been designed and implemented.
Summary · context
The expression system was tested on the anticancer peptides p53pAnt and PNC27.
In this study, PNC-27 was a lab tool or a carrier, part of a mix, or the subject of a lab method. This result is not evidence about taking PNC-27.
Show in the study text Report a problemFND-2488524202

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Version history

PepBank keeps every version of the data on this study. Status changes from Retraction Watch, Europe PMC and Crossref are listed with their source.

Version 2Current
21 Oct 2026, 15:58 UTC
Study results updated
Version 1
21 Oct 2026, 12:50 UTC
First added · source update from Europe PMC

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