Evidence map›Paper›PMID 35089557›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2022

Two-Tiered Selection and Screening Strategy to Increase Functional Enzyme Production in E. coli.

Jason T Boock, May Taw, Brian C King, Robert J Conrado, Donna M Gibson, Matthew P DeLisa

Abstract read
PubMed Publisher
In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
0.5field-weighted citation impact, top 47% 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

0 citing papers in PubMed, 1 citations in OpenAlex.

No citing paper in PubMed yet.

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

6 authors at 2 institutions in 1 country.

Jason T BoockRobert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA. boockj@miamioh.edu.
May TawDepartment of Microbiology, Cornell University, Ithaca, NY, USA.
Brian C KingDepartment of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, NY, USA.
Robert J ConradoRobert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA.
Donna M GibsonDepartment of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, NY, USA.
Matthew P DeLisaRobert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY, USA.
Cornell University · USPlant (United States) · US

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Development of recombinant enzymes as industrial biocatalysts or metabolic pathway elements requires soluble expression of active protein. Here we present a two-step strategy, combining a directed evolution selection with an enzyme activity screen, to increase the soluble production of enzymes in the cytoplasm of E. coli. The directed evolution component relies on the innate quality control of the twin-arginine translocation pathway coupled with antibiotic selection to isolate point mutations that promote intracellular solubility. A secondary screen is applied to ensure the solubility enhancement has not compromised enzyme activity. This strategy has been successfully applied to increase the soluble production of a fungal endocellulase by 30-fold in E. coli without change in enzyme specific activity through two rounds of directed evolution.

Indexed as

Escherichia coliSolubilityDirected evolutionEnzyme expressionHigh-throughput screenTwin-arginine translocation pathway

Identifiers

PMID35089557
OpenAlexW4210275800

What OpenQuestion holds

Textmetadata
Read underepoch 390

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.