ArticleApplied and environmental microbiology2003
Improvement of posttranslational bottlenecks in the production of penicillin amidase in recombinant Escherichia coli strains.
Article in Applied and environmental microbiology, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
15 citing papers in PubMed, 53 citations in OpenAlex.
- Systematic mining and engineering of signal peptides to achieve efficient secretion of cephalosporin C acylase inSynthetic and systems biotechnology · 2026Article
- Cellulase secretion by engineered Pseudomonas putida enables growth on cellulose oligomers.Applied microbiology and biotechnology · 2025Article
- Optimizing the Expression and Solubilization of an E. coli-Produced Leukemia Inhibitory Factor for Anti-LIF Antibody Production and Use Thereof for Contraception in Mice.Molecular biotechnology · 2021Article
- Exploitation of E. coli for the production of penicillin G amidase: a tool for the synthesis of semisynthetic β-lactam antibiotics.Journal, genetic engineering & biotechnology · 2021Review
- Developing Gram-negative bacteria for the secretion of heterologous proteins.Microbial cell factories · 2018Review
- Secretion of recombinant proteins fromEngineering in life sciences · 2018Review
- Expression of a fungal manganese peroxidase in Escherichia coli: a comparison between the soluble and refolded enzymes.BMC biotechnology · 2016Article
- High-throughput strategies for penicillin G acylase production in rE. coli fed-batch cultivations.BMC biotechnology · 2014Article
- Recombinant expression and functional analysis of proteases from Streptococcus pneumoniae, Bacillus anthracis, and Yersinia pestis.BMC biochemistry · 2011Article
- Improvement of penicillin G acylase expression in Escherichia coli through UV induced mutations.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2010Article
- Protein engineering of penicillin acylase.Acta naturae · 2010Article
- Strategies for successful recombinant expression of disulfide bond-dependent proteins in Escherichia coli.Microbial cell factories · 2009Article
- Use of folding modulators to improve heterologous protein production in Escherichia coli.Microbial cell factories · 2009Article
- Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies.Molecular and cellular biochemistry · 2008Review
- Improvement of an unusual twin-arginine transporter leader peptide by a codon-based randomization approach.Applied and environmental microbiology · 2006Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Using periplasmic penicillin amidase (PA) from Escherichia coli ATCC 11105 as a model recombinant protein, we reviewed the posttranslational bottlenecks in its overexpression and undertook attempts to enhance its production in different recombinant E. coli expression hosts. Intracellular proteolytic degradation of the newly synthesized PA precursor and translocation through the plasma membrane were determined to be the main posttranslational processes limiting enzyme production. Rate constants for both intracellular proteolytic breakdown (k(d)) and transport (k(t)) were used as quantitative tools for selection of the appropriate host system and cultivation medium. The production of mature active PA was increased up to 10-fold when the protease-deficient strain E. coli BL21(DE3) was cultivated in medium without a proteinaceous substrate, as confirmed by a decrease in the sum of the constants k(d) and k(t). The original signal sequence of pre-pro-PA was exchanged with the OmpT signal peptide sequence in order to increase translocation efficiency; the effects of this change varied in the different E. coli host strains. Furthermore, we established that simultaneous coexpression of the OmpT pac gene with some proteins of the Sec export machinery of the cell resulted in up to threefold-enhanced PA production. In parallel, we made efforts to increase PA flux via coexpression with the kil gene (killing protein). The primary effects of the kil gene were the release of PA into the extracellular medium and an approximately threefold increase in the total amount of PA produced per liter of bacterial culture.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.