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Related Concept Videos

Amines to Amides: Acylation of Amines01:19

Amines to Amides: Acylation of Amines

Various carboxylic acid derivatives (such as acid chlorides, esters, and anhydrides) can be used for the acylation of amines to yield amides. The reaction requires two equivalents of amines. The first amine molecule functions as a nucleophile and attacks the carbonyl carbon to produce a tetrahedral intermediate. This is followed by the loss of the leaving group and restoration of the C=O bond.
Next, the second equivalent of amine serves as a Brønsted base and deprotonates the quaternary amide...
Preparation of Amines: Reductive Amination of Aldehydes and Ketones01:38

Preparation of Amines: Reductive Amination of Aldehydes and Ketones

Carbonyl compounds and primary amines undergo reductive amination first to produce imines, followed by secondary amines in the same reaction mixture, using selective reducing agents like sodium cyanoborohydride or sodium triacetoxyborohydride. Reductive amination produces different degrees of substitution of amines depending on the starting amine substrate.
Preparation of 1° Amines: Gabriel Synthesis01:28

Preparation of 1° Amines: Gabriel Synthesis

Direct alkylation is not a suitable method for synthesizing amines because it produces polyalkylated products. Gabriel synthesis is the most preferred method to exclusively make primary amines. The method uses phthalimide, which contains a protected form of nitrogen that participates in alkylation only once to predominantly give primary amines.
Strong bases like NaOH or KOH deprotonate the phthalimide to form the corresponding anion, which acts as a nucleophile. Further, the anion attacks an...
Amines to Alkenes: Hofmann Elimination01:16

Amines to Alkenes: Hofmann Elimination

Alkenes can be obtained from amines via an E2 elimination. The amine is first converted into a good leaving group, such as a quaternary ammonium salt. This is accomplished by treating the amine with an excess of alkyl halide, which results in a halide salt. Next, the halide salt is transformed into a hydroxide salt that functions as a base to enable elimination.
Under thermal conditions, the hydroxide can abstract a proton from the β carbon; this generates an alkene with the simultaneous...
Carboxylic Acids to Esters: Acid-Catalyzed (Fischer) Esterification Mechanism01:13

Carboxylic Acids to Esters: Acid-Catalyzed (Fischer) Esterification Mechanism

Carboxylic acids react with alcohols to yield esters via an acid-catalyzed condensation reaction called Fischer esterification. This is a nucleophilic acyl substitution reaction that proceeds via a tetrahedral intermediate, where a water molecule is eliminated as the leaving group.
Preparation of 1° Amines: Hofmann and Curtius Rearrangement Mechanism01:26

Preparation of 1° Amines: Hofmann and Curtius Rearrangement Mechanism

The Hofmann and Curtius rearrangement reactions can be applied to synthesize primary amines from carboxylic acid derivatives such as amides and acyl azides. In the Hofmann rearrangement, a primary amide undergoes deprotonation in the presence of a base, followed by halogenation to generate an N-haloamide. A second proton abstraction produces a stabilized anionic species, which rearranges to an isocyanate intermediate via an alkyl group migration from the carbonyl carbon to the neighboring...

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Related Experiment Video

Updated: Jun 19, 2026

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
10:21

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

Published on: June 20, 2019

A programmable bifunctional flavoenzyme for direct amine-to-ester conversion.

Yaoyun Wu1,2,3, Wei Song1,3, Wanqing Wei2,3

  • 1School of Life Sciences and Health Engineering, Jiangnan University, Wuxi 214122, China.

Science Advances
|June 17, 2026
PubMed
Summary
This summary is machine-generated.

Researchers engineered a novel ancestral flavoenzyme, AncFO-221, to merge amine oxidation and Baeyer-Villiger oxidation (AO-BVO) in one active site. This bifunctional biocatalyst enables direct amine-to-ester conversion, overcoming a major challenge in enzyme design.

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

Published on: February 16, 2018

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Last Updated: Jun 19, 2026

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

Published on: June 20, 2019

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade
09:50

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade

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Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine
09:14

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

Published on: February 16, 2018

Area of Science:

  • Biocatalysis
  • Enzyme Engineering
  • Ancestral Protein Reconstruction

Background:

  • Unifying mechanistically distinct oxidative transformations in a single enzyme active site is a significant challenge.
  • Flavin-dependent oxidative deamination and Baeyer-Villiger oxidation have historically been mechanistically segregated.
  • Integrating these reactions raises questions about potential interference within a single active site.

Purpose of the Study:

  • To design and reconstruct a bifunctional ancestral flavoenzyme capable of direct amine-to-ester conversion.
  • To investigate the mechanistic basis for integrating amine oxidation and Baeyer-Villiger oxidation.
  • To engineer an enhanced variant with improved catalytic efficiency and tunable regioselectivity.

Main Methods:

  • Function-oriented ancestral sequence reconstruction to create AncFO-221.
  • Combined experimental and computational analyses to elucidate the catalytic mechanism.
  • Modular protein engineering to optimize enzyme performance and regioselectivity.

Main Results:

  • Successfully reconstructed AncFO-221, a bifunctional enzyme enabling direct amine-to-ester conversion.
  • Revealed a unified catalytic framework coupling amine oxidation and Baeyer-Villiger oxidation (AO-BVO) via histidine-assisted proton/hydride transfer and C4a-peroxyflavin.
  • Engineered variant M15 showed an 18-fold increase in efficiency, 93% lactone yield, and programmable regioselectivity overriding the classical migratory rule.

Conclusions:

  • Ancestral reconstruction can merge evolutionarily segregated chemistries into a single, tunable catalytic platform.
  • The study provides a blueprint for designing multistep oxidative biocatalysts.
  • Engineered enzymes can exhibit programmable and unconventional regioselectivity, expanding biocatalytic possibilities.