ArticleNature methods2017
Nontargeted in vitro metabolomics for high-throughput identification of novel enzymes in Escherichia coli.
Article in Nature methods, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 65 papers.
What it found
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Who cites it
65 citing papers in PubMed.
- Revisiting vitamin B6 metabolism: The emerging role of pyridoxal reductase in the pyridoxal 5'-phosphate salvage pathway.The FEBS journal · 2026Review
- Systematic discovery of enzyme promiscuity in Escherichia coli using in vitro metabolomics.Communications biology · 2026Article
- Computational approaches to enzymatic reaction assignment: a review of methods, validations, and future directions.Briefings in bioinformatics · 2025Review
- A combined pseudouridine biomanufacturing platform enabled by a streamlined designer pathway.Nature communications · 2025Article
- Enzyme promiscuity in the field of synthetic biology applied to white biotechnology: Opportunities and weaknesses.Biodesign research · 2025Review
- N- to C-Glycoside Rearrangement of Uridine 5'-Phosphate in Two Enzymatic Steps for the Production of Pseudouridine 5'-Phosphate.Biotechnology and bioengineering · 2025Article
- Coapplication of Adenine with Inosine or Guanosine Supports Rapid ATP Restoration by ATP-deprived Cultured Primary Astrocytes.Neurochemical research · 2025Article
- Inhibitory Mechanisms of Vine Tea Extract and Dihydromyricetin AgainstFoods (Basel, Switzerland) · 2025Article
- Charting unknown metabolic reactions by mass spectrometry-resolved stable-isotope tracing metabolomics.Nature communications · 2025Article
- Activating the d-Tagatose Production Capacity ofJournal of agricultural and food chemistry · 2025Article
- MtlD as a therapeutic target for intestinal and systemic bacterial infections.Journal of bacteriology · 2025Article
- Plasmid Library Construction From Genomic DNA.Current protocols · 2025Article
- Review
- The Y-ome Conundrum: Insights into Uncharacterized Genes and Approaches for Functional Annotation.Molecular and cellular biochemistry · 2024Review
- Emerging tools and best practices for studying gut microbial community metabolism.Nature metabolism · 2024Review
- Back to the Future of Metabolism-Advances in the Discovery and Characterization of Unknown Biocatalytic Functions and Pathways.Life (Basel, Switzerland) · 2024Review
- Systematic Engineering ofACS omega · 2024Article
- Functional and structural properties of pyridoxal reductase (PdxI) from Escherichia coli: a pivotal enzyme in the vitamin B6 salvage pathway.The FEBS journal · 2023Article
- Discovery of a non-canonical prototype long-chain monoacylglycerol lipase through a structure-based endogenous reaction intermediate complex.Nature communications · 2023Article
- UHPLC/MS-Based Untargeted Metabolomics Reveals Metabolic Characteristics of Clinical Strain ofMicroorganisms · 2023Article
5 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Our understanding of metabolism is limited by a lack of knowledge about the functions of many enzymes. Here, we develop a high-throughput mass spectrometry approach to comprehensively profile proteins for in vitro enzymatic activity. Overexpressed or purified proteins are incubated in a supplemented metabolome extract containing hundreds of biologically relevant candidate substrates, and accumulating and depleting metabolites are determined by nontargeted mass spectrometry. By combining chemometrics and database approaches, we established an automated pipeline for unbiased annotation of the functions of novel enzymes. In screening all 1,275 functionally uncharacterized Escherichia coli proteins, we discovered 241 potential novel enzymes, 12 of which we experimentally validated. Our high-throughput in vitro metabolomics method is generally applicable to any purified protein or crude cell lysate of its overexpression host and enables performing up to 1,200 nontargeted enzyme assays per working day.
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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.