Related Experiment Video

Updated: Apr 15, 2026

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Published on: June 20, 2019

α-Carbonic Anhydrases Possess Thioesterase Activity

Muhammet Tanc¹, Fabrizio Carta², Andrea Scozzafava²

¹Laboratorio di Chimica Bioinorganica, Università degli Studi di Firenze, Polo Scientifico , Room 188, Via della Lastruccia 3, 50019 Sesto Fiorentino, Florence, Italy ; NEUROFARBA Department, Sezione di Scienze Farmaceutiche e Nutraceutiche, Università degli Studi di Firenze , Via Ugo Schiff 6, 50019 Sesto Fiorentino, Florence, Italy.

ACS Medicinal Chemistry Letters

|March 28, 2015

Summary

Alpha-carbonic anhydrases (CAs) exhibit esterase activity and, as shown here, also possess thioesterase activity. This newly discovered CA function may aid in developing novel enzyme inhibitors and drugs with multiple mechanisms of action.

Area of Science:

Biochemistry
Enzymology
Medicinal Chemistry

Background:

Alpha-carbonic anhydrases (CAs) are known for their esterase activity on various ester types.
Their full catalytic repertoire is continually being explored to uncover novel biochemical functions.

Purpose of the Study:

To investigate the potential thioesterase activity of alpha-carbonic anhydrases.
To characterize the hydrolysis products and mechanism of CA-mediated thioester hydrolysis.

Main Methods:

Kinetic studies
Spectroscopic analyses
Mass spectrometry (MS) to identify hydrolysis products.

Main Results:

Demonstrated that alpha-carbonic anhydrases possess thioesterase activity using a dithiocarbamate ester substrate (PhSO2NHCSSMe).
Identified benzenesulfonamide, methyl mercaptan, and COS as hydrolysis products.
Confirmed CA-mediated hydrolysis through kinetic, spectroscopic, and MS data.

Conclusions:

Alpha-carbonic anhydrases exhibit previously unrecognized thioesterase activity.
This activity could be leveraged for designing prodrug enzyme inhibitors.
CA thioesterase function may play a role in physiological/pathological processes, offering avenues for drug discovery targeting multiple mechanisms.

Keywords:

Carbonic anhydrase inhibitors prodrugs thioesterase

More Related Videos

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine

Published on: February 16, 2018

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Published on: November 23, 2016

Related Concept Videos

Amides to Carboxylic Acids: Hydrolysis

Amides to Carboxylic Acids: Hydrolysis

Amides can undergo either acid-catalyzed hydrolysis or base-promoted hydrolysis through a typical nucleophilic acyl substitution. Each hydrolysis requires severe conditions.
Acid-catalyzed hydrolysis:
Hydrolysis of amides under acidic conditions yields carboxylic acids. Since the reaction occurs slowly, hydrolysis requires the conditions of heat.
The mechanism begins with the protonation of the carbonyl oxygen by the acid catalyst. The protonation makes the amide carbonyl carbon more...

Regioselectivity and Stereochemistry of Acid-Catalyzed Hydration

Regioselectivity and Stereochemistry of Acid-Catalyzed Hydration

The rate of acid-catalyzed hydration of alkenes depends on the alkene's structure, as the presence of alkyl substituents at the double bond can significantly influence the rate.

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...

Nitriles to Carboxylic Acids: Hydrolysis

Nitriles to Carboxylic Acids: Hydrolysis

Nitriles undergo acid-catalyzed hydrolysis or base-catalyzed hydrolysis to form a carboxylic acid. These reactions proceed via an amide intermediate.

Acid Halides to Carboxylic Acids: Hydrolysis

Acid Halides to Carboxylic Acids: Hydrolysis

Hydrolysis of acid halides is a nucleophilic acyl substitution reaction in which acid halides react with water to give carboxylic acids. The reaction occurs readily and does not require acid or a base catalyst.
As shown below, the mechanism involves a nucleophilic attack by water at the carbonyl carbon to form a tetrahedral intermediate. This is followed by the reformation of the carbon–oxygen π bond along with the departure of a halide ion. A final proton transfer step yields...

Loss of Carboxy Group as CO2: Decarboxylation of β-Ketoacids

Loss of Carboxy Group as CO₂: Decarboxylation of β-Ketoacids

Carboxylic acids, upon heating, undergo a decarboxylation reaction by releasing carbon dioxide gas. Monocarboxylic acids do not undergo decarboxylation easily. However, a silver salt of carboxylic acid reacts with bromine or iodine under high temperature to release carbon dioxide gas and forms halide with one less carbon. This reaction is called the Hunsdiecker reaction.

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

Treatment of NSAPs-rich petrochemical wastewaters using a two-stage combined process of fungi and activated sludge.

Environmental technology·2020

Dioxygen, an unexpected carbonic anhydrase ligand.

Journal of enzyme inhibition and medicinal chemistry·2018

Discovery of Novel Nonsteroidal Anti-Inflammatory Drugs and Carbonic Anhydrase Inhibitors Hybrids (NSAIDs-CAIs) for the Management of Rheumatoid Arthritis.

Journal of medicinal chemistry·2018

Sulphonamide inhibition studies of the β-carbonic anhydrase from the bacterial pathogen Clostridium perfringens.

Journal of enzyme inhibition and medicinal chemistry·2017

1,3-Oxazole-based selective picomolar inhibitors of cytosolic human carbonic anhydrase II alleviate ocular hypertension in rabbits: Potency is supported by X-ray crystallography of two leads.

Bioorganic & medicinal chemistry·2017

Carbonic anhydrase activation enhances object recognition memory in mice through phosphorylation of the extracellular signal-regulated kinase in the cortex and the hippocampus.

Neuropharmacology·2017

See all related articles