Evidence map›Paper›PMID 40368116›Full record

ReviewBiotechnology advances2025

Protease engineering: Approaches, tools, and emerging trends.

Samantha G Martinusen, Sage E Nelson, Ethan W Slaton, Lawton F Long, Raymond Pho, Seyednima Ajayebi, Carl A Denard

Abstract readReview
PubMed Full text at PMC Publisher
In one paragraph

Review in Biotechnology advances, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.

0numbers the graph read from it
0cells of the map it votes in
7citing papers in PubMed
–field-weighted citation impact
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

7 citing papers in PubMed.

  1. Review
  2. Article
  3. Review
  4. Multicyclic D-Stereospecific Hydrolase Dimer With High Sustained Activity.Angewandte Chemie (International ed. in English) · 2026
    Article
  5. Review
  6. Review
  7. 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.

Samantha G MartinusenDepartment of Chemical Engineering, University of Florida, Gainesville 32611, USA.
Sage E NelsonDepartment of Chemical Engineering, University of Florida, Gainesville 32611, USA.
Ethan W SlatonDepartment of Chemical Engineering, University of Florida, Gainesville 32611, USA.
Lawton F LongDepartment of Chemical Engineering, University of Florida, Gainesville 32611, USA.
Raymond PhoDepartment of Chemical Engineering, University of Florida, Gainesville 32611, USA.
Seyednima AjayebiDepartment of Chemical Engineering, University of Florida, Gainesville 32611, USA.
Carl A DenardDepartment of Chemical Engineering, University of Florida, Gainesville 32611, USA; UF Health Cancer Center, University of Florida, Gainesville, 32611, USA. Electronic address: cdenard@ufl.edu.

Funding

Reprogramming proteases: tackling human diseases with next-generation modulatorsR35GM146821 · NIGMS · UNIVERSITY OF FLORIDA · PI Carl Denard · 2022 to 2026
$1.6M
NIGMS NIH HHS R35 GM146821
6 · The paper itself

Abstract

Engineered proteases with bespoke substrate specificities and activities can empower broad and innovative applications in biomedicine, mass spectrometry-based proteomics, and chemical and synthetic biology. This review provides an authoritative, topical, and detailed description and discussion of the directed evolution and high-throughput strategies designed to engineer the substrate specificity of proteases in E. coli, yeast, phage, and cell-free systems. Second, we discuss emerging protease engineering strategies that complement directed evolution, including antibody-protease fusions that enable proximity catalysis, and protease substrate specificity switching driven by exogenous protein-protein interactions. Lastly, we discuss principles for engineering split and autoinhibited proteases, which are key signal-processing modules in protein circuits. Overall, readers will gain a valuable understanding of the latest advances in protease engineering, focusing on methodologies and strategies that enable precise control of protease activity and specificity.

Indexed as

Peptide HydrolasesProtein EngineeringDirected Molecular EvolutionHumansSubstrate SpecificityPeptide HydrolasesCell-freeHigh-throughput screeningMachine learning learningProtease substrate specificity engineering directed evolution antibody

Identifiers

PMID40368116
PMCPMC12381794

What OpenQuestion holds

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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.