Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Alkylation of β-Diester Enolates: Malonic Ester Synthesis

Alkylation of β-Diester Enolates: Malonic Ester Synthesis

Malonic ester synthesis is a method to obtain α substituted carboxylic acids from ꞵ-diesters such as diethyl malonate and alkyl halides.

Phosphodiester Linkages

Phosphodiester Linkages

Overview
Phosphodiester bond forms when a phosphoric acid molecule (H3PO4) links with two hydroxyl groups (–OH) of two other molecules, forming two ester bonds. Two water molecules are released in this process. The phosphodiester bond is commonly found in nucleic acids (DNA and RNA) and plays a critical role in their structure and function.
Phosphodiester Bonds Link Nucleotides Together
DNA and RNA are polynucleotides or long chains of nucleotides that are linked together. A nucleotide is...

Restriction Enzymes

Restriction Enzymes

Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...

Esters to Carboxylic Acids: Acid-Catalyzed Hydrolysis

Esters to Carboxylic Acids: Acid-Catalyzed Hydrolysis

Hydrolysis of esters under acidic conditions proceeds through a nucleophilic acyl substitution. In the presence of excess water, the reaction proceeds in a reversible manner, forming carboxylic acids and alcohols.
During hydrolysis, the ester is first activated towards nucleophilic attack through the protonation of the carboxyl oxygen atom by the acid catalyst. The protonation makes the ester carbonyl carbon more electrophilic. In the next step, water acts as a nucleophile and adds to the...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Elucidation of the Pathway for the Biosynthesis of the Undecorated Capsular Polysaccharide from the HS:19 Serotype of the Human Pathogen <i>Campylobacter jejuni</i>.

Biochemistry·2026

Same author

Metabolic thermodynamics: pertinent reference state and energy potentials.

The FEBS journal·2026

Same author

Light-Triggered Accelerated Degradation of Surface-Bound Chemical Agents.

Journal of the American Chemical Society·2026

Same author

Enzymatic Synthesis of an Undecorated Capsular Polysaccharide from <i>Campylobacter jejuni</i>.

Biochemistry·2025

Same author

Biosynthesis of CDP-α-d-fucofuranose and CDP-β-l-6-deoxy-altrofuranose for the Capsular Polysaccharides of <i>Campylobacter jejuni</i>.

Biochemistry·2025

Same author

The use of phosphotriesterase in the synthesis of enantiomerically pure ProTide prodrugs.

Chemico-biological interactions·2025

Same journal

Aromatic Cage-Directed Azide-Methyllysine Photochemistry for Profiling Nonhistone Interacting Partners of the MeCP2 Methyl-CpG-Binding Domain.

Biochemistry·2026

Same journal

Differential Hydroxypyruvate Processing by <i>E. coli</i> and <i>P. aeruginosa</i> DXP Synthases Reveals Preferential Xylulose 5-Phosphate Formation by the <i>P. aeruginosa</i> Enzyme.

Biochemistry·2026

Same journal

Structural and Functional Characterization of Heterologous Nitrogenase Complexes.

Biochemistry·2026

Same journal

Discovery of Bacterial Unspecific Peroxygenases.

Biochemistry·2026

Same journal

Lactate Biology: Subcellular Routing and Chemical Form Define Function.

Biochemistry·2026

Same journal

Nature's Anaerobic Toolkit: Glycyl Radical Enzymes and Their Expanding Functional and Mechanistic Diversity.

Biochemistry·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Apr 14, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

Function discovery and structural characterization of a methylphosphonate esterase.

Dao Feng Xiang, Yury Patskovsky¹, Venkatesh V Nemmara

¹‡Department of Biochemistry, Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York, 10461, United States.

|April 16, 2015

Summary

This summary is machine-generated.

The enzyme Pmi1525 efficiently hydrolyzes organophosphonate and carboxylate esters, showing stereoselectivity for chiral methylphosphonate esters. Its catalytic properties offer potential for developing new enzymes for nerve agent detoxification.

More Related Videos

In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity

In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity

Published on: January 5, 2016

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Published on: August 8, 2016

Related Experiment Videos

Last Updated: Apr 14, 2026

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity

In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity

Published on: January 5, 2016

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Published on: August 8, 2016

Area of Science:

Biochemistry
Enzymology
Structural Biology

Background:

Pmi1525 is an enzyme from Proteus mirabilis HI4320 with an unknown function.
It belongs to the amidohydrolase superfamily, characterized by a (β/α)8-barrel fold.

Purpose of the Study:

To clone, purify, and functionally characterize Pmi1525.
To determine the three-dimensional structure of Pmi1525 with bound ligands.
To identify the substrate profile and catalytic properties of Pmi1525.

Main Methods:

Gene cloning and protein purification.
X-ray crystallography for structure determination (PDB IDs: 3RHG, 4QSF).
Bioinformatics, structural characterization, and focused library screening for substrate profiling.

Main Results:

Pmi1525 efficiently hydrolyzes organophosphonate and carboxylate esters.
Optimal substrates include ethyl 4-nitrophenylmethyl phosphonate and 4-nitrophenyl butyrate.
The enzyme exhibits stereoselectivity, hydrolyzing the (SP)-enantiomer of isobutyl 4-nitrophenyl methylphosphonate 14 times faster than the (RP)-enantiomer.

Conclusions:

Pmi1525 is a functional amidohydrolase with a defined substrate profile.
Its catalytic properties make it a promising template for evolving enzymes for organophosphonate detection and detoxification.
Structural insights provide a basis for understanding its mechanism and engineering new functionalities.