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Study · Not a test of the peptide alone · 2011

5 results

Production of Nα-acetylated thymosin α1 in Escherichia coli.

Ren Y, Yao X, Dai H, Li S, Fang H, Chen H, Zhou C

Microbial cell factories22 Apr 2011Cited by 18 (Europe PMC)

PMID21513520 (opens in a new tab)PMCIDPMC3103413 (opens in a new tab)DOI10.1186/1475-2859-10-26 (opens in a new tab)PepBankPUB-21513520
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Summary

BackgroundThymosin α1 (Tα1), a 28-amino acid Nα-acetylated peptide, has a powerful general immunostimulating activity. Although biosynthesis is an attractive means of large-scale manufacture, to date, Tα1 can only be chemosynthesized because of two obstacles to its biosynthesis: the difficulties in expressing small peptides and obtaining Nα-acetylation. In this study, we describe a novel production process for Nα-acetylated Tα1 in Escherichia coli.ResultsTo obtain recombinant Nα-acetylated Tα1 efficiently, a fusion protein, Tα1-Intein, was constructed, in which Tα1 was fused to the N-terminus of the smallest mini-intein, Spl DnaX (136 amino acids long, from Spirulina platensis), and a His tag was added at the C-terminus. Because Tα1 was placed at the N-terminus of the Tα1-Intein fusion protein, Tα1 could be fully acetylated when the Tα1-Intein fusion protein was co-expressed with RimJ (a known prokaryotic Nα-acetyltransferase) in Escherichia coli. After purification by Ni-Sepharose affinity chromatography, the Tα1-Intein fusion protein was induced by the thiols β-mercaptoethanol or d,l-dithiothreitol, or by increasing the temperature, to release Tα1 through intein-mediated N-terminal cleavage. Under the optimal conditions, more than 90% of the Tα1-Intein fusion protein was thiolyzed, and 24.5 mg Tα1 was obtained from 1 L of culture media. The purity was 98% after a series of chromatographic purification steps. The molecular weight of recombinant Tα1 was determined to be 3107.44 Da by mass spectrometry, which was nearly identical to that of the synthetic version (3107.42 Da). The whole sequence of recombinant Tα1 was identified by tandem mass spectrometry and its N-terminal serine residue was shown to be acetylated.ConclusionsThe present data demonstrate that Nα-acetylated Tα1 can be efficiently produced in recombinant E. coli. This bioprocess could be used as an alternative to chemosynthesis for the production of Tα1. The described methodologies may also be helpful for the biosynthesis of similar peptides.
CC BYText from Europe PMC. Highlighted sentences are the source of results.

Results from this study5

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

Studies that did not test the peptide alone5

In this study, Thymosin alpha-1 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 Thymosin alpha-1.

  1. 01LabNot a test of Thymosin alpha-1 aloneIn vitro (cell-free)Other

    Under optimal conditions, more than 90% of the Tα1-Intein fusion protein was thiolyzed to release thymosin alpha‑1.

    In plain wordsIn lab production, over 90% of the fusion protein was cleaved to release thymosin alpha‑1.

    Peptide
    Thymosin alpha-1
    Outcome
    Not applicableIntein-mediated cleavage of Tα1-Intein
    Study

    n: not reported in source

    Recombinant expression in Escherichia coli
    Outcome group
    Synthesis and manufacturing · Pharmacology

    Measurements

    Fusion protein thiolyzed
    > 90%
    optimal conditions

    Source sentences

    Summary
    Under the optimal conditions, more than 90% of the Tα1-Intein fusion protein was thiolyzed
    1 context sentence
    Summary · context
    In this study, we describe a novel production process for Nα-acetylated Tα1 in Escherichia coli.

    In this study, Thymosin alpha-1 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 Thymosin alpha-1.

  2. 02LabNot a test of Thymosin alpha-1 aloneIn vitro (cell-free)Other

    The process gave 24.5 mg of thymosin alpha‑1 from 1 L of culture media.

    In plain wordsIn lab production, 24.5 mg of thymosin alpha‑1 came from 1 litre of culture.

    Peptide
    Thymosin alpha-1
    Outcome
    Not applicableTα1 yield from E. coli culture
    Study

    n: not reported in source

    Recombinant expression in Escherichia coli
    Outcome group
    Synthesis and manufacturing · Pharmacology

    Measurements

    Yield
    24.5 mg/L
    culture media

    Source sentences

    Summary
    24.5 mg Tα1 was obtained from 1 L of culture media.
    1 context sentence
    Summary · context
    In this study, we describe a novel production process for Nα-acetylated Tα1 in Escherichia coli.

    In this study, Thymosin alpha-1 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 Thymosin alpha-1.

  3. 03LabNot a test of Thymosin alpha-1 aloneIn vitro (cell-free)Other

    Recombinant thymosin alpha‑1 reached 98% purity after chromatographic purification.

    In plain wordsIn lab production, the purified thymosin alpha‑1 was 98% pure.

    Peptide
    Thymosin alpha-1
    Outcome
    Not applicablePurity of recombinant Tα1
    Study

    n: not reported in source

    Recombinant expression in Escherichia coli
    Outcome group
    Synthesis and manufacturing · Pharmacology

    Measurements

    Purity
    98%
    recombinant Tα1

    Source sentences

    Summary
    The purity was 98% after a series of chromatographic purification steps.
    1 context sentence
    Summary · context
    In this study, we describe a novel production process for Nα-acetylated Tα1 in Escherichia coli.

    In this study, Thymosin alpha-1 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 Thymosin alpha-1.

  4. 04LabNot a test of Thymosin alpha-1 aloneIn vitro (cell-free)Other

    Mass spectrometry gave 3107.44 Da for recombinant thymosin alpha‑1, nearly identical to the synthetic version (3107.42 Da).

    In plain wordsIn lab tests, recombinant thymosin alpha‑1 had almost the same mass as the synthetic peptide.

    Peptide
    Thymosin alpha-1
    Outcome
    Not applicableMolecular weight of recombinant Tα1
    Study

    n: not reported in source

    Recombinant expression in Escherichia coli
    Outcome group
    Synthesis and manufacturing · Pharmacology

    Measurements

    Molecular weight
    3107.44 Da
    recombinant Tα1
    Molecular weight
    3107.42 Da
    synthetic Tα1

    Source sentences

    Summary
    The molecular weight of recombinant Tα1 was determined to be 3107.44 Da by mass spectrometry, which was nearly identical to that of the synthetic version (3107.42 Da).
    1 context sentence
    Summary · context
    In this study, we describe a novel production process for Nα-acetylated Tα1 in Escherichia coli.

    In this study, Thymosin alpha-1 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 Thymosin alpha-1.

  5. 05LabNot a test of Thymosin alpha-1 aloneIn vitro (cell-free)Other

    Tandem mass spectrometry identified the full sequence of recombinant thymosin alpha‑1 and showed an acetylated N-terminal serine.

    In plain wordsIn lab tests, recombinant thymosin alpha‑1 carried the expected acetyl group on its first amino acid.

    Peptide
    Thymosin alpha-1
    Outcome
    Not applicableNα-acetylation of recombinant Tα1
    Study

    n: not reported in source

    Recombinant expression in Escherichia coli
    Outcome group
    Synthesis and manufacturing · Pharmacology

    Source sentences

    Summary
    The whole sequence of recombinant Tα1 was identified by tandem mass spectrometry and its N-terminal serine residue was shown to be acetylated.
    1 context sentence
    Summary · context
    In this study, we describe a novel production process for Nα-acetylated Tα1 in Escherichia coli.

    In this study, Thymosin alpha-1 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 Thymosin alpha-1.

AI wrote and checked these results. AI can make mistakes, so check important results against the source.

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.

  1. Version 3Current

    10 Oct 2026, 08:55 UTC

    Study results updated

  2. Version 2

    9 Oct 2026, 03:03 UTC

    Study data updated · source update from Europe PMC

  3. Version 1

    9 Oct 2026, 03:02 UTC

    First added · source update from Europe PMC

Subjects

Terms as the source lists them.

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