Evidence map›Paper›PMID 37405436›Full record

ArticleApplied microbiology and biotechnology2023

A step forward to the optimized HlyA type 1 secretion system through directed evolution.

Zohreh N Pourhassan, Haiyang Cui, Neele Muckhoff, Mehdi D Davari, Sander H J Smits, Ulrich Schwaneberg, Lutz Schmitt

Open access · hybridAbstract read
In one paragraph

Article in Applied microbiology and biotechnology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
1.5field-weighted citation impact, top 18% of its field
1 · What the graph read from it

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.

2 · The registry

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.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed, 6 citations in OpenAlex.

  1. Article
  2. Review
  3. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors at 4 institutions in 2 countries.

Zohreh N PourhassanInstitute of Biochemistry, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.ORCID http://orcid.org/0000-0002-9340-9619
Haiyang CuiInstitute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074, Aachen, Germany.ORCID http://orcid.org/0000-0001-8360-0447
Neele MuckhoffInstitute of Biochemistry, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.ORCID http://orcid.org/0000-0002-2471-7116
Mehdi D DavariDepartment of Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle, Germany.ORCID http://orcid.org/0000-0003-0089-7156
Sander H J SmitsInstitute of Biochemistry, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany.ORCID http://orcid.org/0000-0003-0780-9251
Ulrich SchwanebergInstitute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074, Aachen, Germany.ORCID http://orcid.org/0000-0003-4026-701X
Lutz SchmittInstitute of Biochemistry, Heinrich Heine University, Universitätsstr. 1, 40225, Düsseldorf, Germany. lutz.schmitt@hhu.de.ORCID http://orcid.org/0000-0002-1167-9819
Heinrich Heine University Düsseldorf · DEDWI – Leibniz Institute for Interactive Materials · DELeibniz Institute of Plant Biochemistry · DEUniversity of Illinois Urbana-Champaign · US

Funding

European Regional Development Fund 34.EFRE-0300096
6 · The paper itself

Abstract

Secretion of proteins into the extracellular space has great advantages for the production of recombinant proteins. Type 1 secretion systems (T1SS) are attractive candidates to be optimized for biotechnological applications, as they have a relatively simple architecture compared to other classes of secretion systems. A paradigm of T1SS is the hemolysin A type 1 secretion system (HlyA T1SS) from Escherichia coli harboring only three membrane proteins, which makes the plasmid-based expression of the system easy. Although for decades the HlyA T1SS has been successfully applied for secretion of a long list of heterologous proteins from different origins as well as peptides, but its utility at commercial scales is still limited mainly due to low secretion titers of the system. To address this drawback, we engineered the inner membrane complex of the system, consisting of HlyB and HlyD proteins, following KnowVolution strategy. The applied KnowVolution campaign in this study provided a novel HlyB variant containing four substitutions (T36L/F216W/S290C/V421I) with up to 2.5-fold improved secretion for two hydrolases, a lipase and a cutinase. KEY POINTS: • An improvement in protein secretion via the use of T1SS • Reaching almost 400 mg/L of soluble lipase into the supernatant • A step forward to making E. coli cells more competitive for applying as a secretion host.

Indexed as

Escherichia coliEscherichia coli ProteinsBacterial ProteinsHemolysin ProteinsLipaseMembrane ProteinsType I Secretion SystemsBacterial ProteinsEscherichia coli ProteinsHemolysin ProteinsLipaseMembrane ProteinsType I Secretion SystemsBacterial secretion systemDirected evolutionGram-negative bacteriaKnowVolutionProtein engineeringProtein secretion

Identifiers

PMID37405436
PMCPMC10386944
OpenAlexW4383186615

What OpenQuestion holds

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LicenceCC BY
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Registered trials

None linked

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.