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

Preparation of 1° Amines: Azide Synthesis

Preparation of 1° Amines: Azide Synthesis

Direct alkylation of ammonia produces polyalkylated amines, along with a quaternary ammonium salt. To exclusively prepare primary amines, the azide synthesis method can be used.
Azide ions act as good nucleophiles and react with unhindered alkyl halides to form alkyl azides. Alkyl azides do not participate in further nucleophilic substitution reactions, thereby eliminating the chances of polyalkylated products. Alkyl azides are reduced by hydride-based reducing agents, like lithium aluminum...

Oxidative Cleavage of Alkenes: Ozonolysis

Oxidative Cleavage of Alkenes: Ozonolysis

In ozonolysis, ozone is used to cleave a carbon–carbon double bond to form aldehydes and ketones, or carboxylic acids, depending on the work-up.
Ozone is a symmetrical bent molecule stabilized by a resonance structure.

Synthetic Biology

Synthetic Biology

Synthetic biology is an interdisciplinary science that involves using principles from disciplines such as engineering, molecular biology, cell biology, and systems biology. It involves remodeling existing organisms from nature or constructing completely new synthetic organisms for applications such as protein or enzyme production, bioremediation, value-added macromolecule production, and the addition of desirable traits to crops, to name a few.
Golden rice
Golden rice is a genetically modified...

Exon Recombination

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon has three reading...

Production of Antibiotics

Production of Antibiotics

Penicillin, one of the earliest and most widely used antibiotics, is produced industrially by the filamentous fungus Penicillium chrysogenum. Large stirred-tank bioreactors ranging from tens to hundreds of thousands of liters maintain tightly controlled temperature, pH, and dissolved oxygen conditions to support fungal metabolism and maximize antibiotic yield. Penicillin is a secondary metabolite, synthesized primarily during the stationary growth phase, which requires a carefully managed...

Biosynthesis in Bacteria

Biosynthesis in Bacteria

Biosynthesis in bacteria is a fundamental anabolic process that generates essential macromolecules, including proteins, nucleic acids, lipids, and polysaccharides. These macromolecules are critical for cellular growth, replication, and function. The process is tightly regulated and energetically linked to catabolic pathways to ensure optimal resource utilization.Biosynthetic pathways begin with precursor metabolites such as pyruvate, acetyl-CoA, and glucose-6-phosphate derived from glycolysis,...

You might also read

Related Articles

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

Sort by

Same author

Structure-Guided Optimization of α-Conotoxins Yields Potent and Selective Inhibitors of the Human α6β4 nAChR for Nicotine Addiction.

Journal of medicinal chemistry·2026

Same author

Computational-Guided Protein Engineering of LaACS Coupled with SAM Synthase Screening Synergistically Boosts ACC Production in <i>E. coli</i>.

Journal of agricultural and food chemistry·2025

Same author

Biocatalytic and Biomimetic Catalysis for Intermolecular Cyclopropanation of Olefins: A Review.

Chemical record (New York, N.Y.)·2025

Same author

Bio-Inspired Consecutive Photocatalyzed C-H Nitration of Arenes.

The Journal of organic chemistry·2025

Same author

Recombinant <i>Escherichia coli</i> Utilizes Mild Hydrogen Sources for the Targeted Intracellular Synthesis of Palladium Nanoparticles and Whole-Cell-Catalyzed Aromatic Aldehyde Hydrogenation.

ACS applied materials & interfaces·2025

Same author

Design Strategy of PepNzymes-SH for an Emerging Catalyst with Serine Hydrolase-Like Functionality.

ACS applied materials & interfaces·2025

Search research articles

Related Experiment Video

Updated: Jun 5, 2026

Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

Published on: November 8, 2019

Developing High Performance N-Oxygenase for Azomycin Synthesis through Ancestral Sequence Reconstruction.

Jinming Dou¹, Bo Qin², Zongjiang Yu³

¹State Key Laboratory Base of Eco-Chemical Engineering, College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao 266101, China.

ACS Synthetic Biology

|June 3, 2026

Summary

Ancestral sequence reconstruction engineered a superior N-oxygenase enzyme, ASR-90, for green azomycin synthesis. This biocatalyst shows enhanced efficiency, stability, and a broad substrate scope, overcoming limitations of traditional methods.

Keywords:

Ancestral sequence reconstruction Azomycin Biocatalysis N-Oxygenase Nitro compounds

More Related Videos

Synthesis of 1,2-Azaborines and the Preparation of Their Protein Complexes with T4 Lysozyme Mutants

Synthesis of 1,2-Azaborines and the Preparation of Their Protein Complexes with T4 Lysozyme Mutants

Published on: March 25, 2017

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition

Published on: October 3, 2018

Related Experiment Videos

Last Updated: Jun 5, 2026

Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

Published on: November 8, 2019

Synthesis of 1,2-Azaborines and the Preparation of Their Protein Complexes with T4 Lysozyme Mutants

Synthesis of 1,2-Azaborines and the Preparation of Their Protein Complexes with T4 Lysozyme Mutants

Published on: March 25, 2017

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition

Published on: October 3, 2018

Area of Science:

Biocatalysis and Enzyme Engineering
Synthetic Biology
Green Chemistry

Background:

Traditional azomycin synthesis involves harsh conditions and low yields.
Biocatalytic routes using N-oxygenase are limited by enzyme efficiency and stability.

Purpose of the Study:

To overcome limitations in biocatalytic N-oxygenase synthesis.
To develop a high-performance N-oxygenase for azomycin production using ancestral sequence reconstruction.

Main Methods:

Resurrected and screened seven ancestral node proteins from 83 homologous sequences.
Utilized ancestral sequence reconstruction (ASR) to identify improved enzyme variants.
Performed molecular dynamics simulations to understand enhanced catalytic mechanisms.

Main Results:

Engineered enzyme ASR-90 demonstrated a 546-fold increase in catalytic efficiency (kcat/Km) for 2-aminoimidazole.
ASR-90 showed improved acid tolerance (optimal pH 4.5) and a 4-fold longer half-life at 50 °C.
Achieved 50.9% substrate conversion in 20 min and displayed catalytic ability on 16 non-natural substrates.

Conclusions:

Ancestral sequence reconstruction is an effective strategy for developing robust N-oxygenases.
ASR-90 facilitates the green biosynthesis of azomycin and its derivatives.
Enhanced substrate binding and active site modifications contribute to ASR-90's superior performance.