Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Restriction Enzymes01:11

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...
Ligand Binding and Linkage00:49

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...

You might also read

Related Articles

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

Sort by
Same author

Directed evolution of a cytochrome P450 monooxygenase for improved perillyl alcohol biosynthesis <i>via</i> a tailored genetically encoded biosensor.

RSC advances·2026
Same author

Engineering an Acyl-CoA Ligase With Enhanced Activity Toward Synthetic CoA Alternatives.

Chembiochem : a European journal of chemical biology·2026
Same author

Development of a transcription factor-based biosensor strain for reporting α-terpineol production <i>via</i> the alcohol-dependent hemiterpene pathway in <i>Escherichia coli</i>.

RSC chemical biology·2026
Same author

Engineering the Specificity of Acetyl-CoA Synthetase for Diverse Acyl-CoA Thioester Generation.

ACS chemical biology·2025
Same author

Promiscuity of an Alcohol-Dependent Hemiterpene Pathway for the In Vivo Production of a Non-Natural Alkylated Tryptophan Derivative.

ACS synthetic biology·2025
Same author

Directed Evolution of a Macrolide-Sensing Transcription Factor Biosensor for the Detection of Macrolactone Aglycones via "Effector Walking" and Efflux Pump Deletion.

Biochemistry·2025

Related Experiment Video

Updated: Jul 12, 2026

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes T&#252;6028
09:08

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028

Published on: January 13, 2017

Dissection and Engineering of Modular Polyketide Synthase Extender Unit Specificity Motifs.

Sydney Welch1, Gavin J Williams1,2

  • 1Department of Chemistry, NC State University, Raleigh, North Carolina, USA.

Chembiochem : a European Journal of Chemical Biology
|July 10, 2026
PubMed
Summary

Redesigning polyketide synthases (PKSs) for new natural products is challenging. This study identifies key sequence motifs in acyltransferase (AT) domains that control extender unit specificity, enabling novel polyketide generation.

Keywords:
acyl‐CoAenzymespolyketide synthasepolyketidessubstrate specificity

More Related Videos

Optimization of Synthetic Proteins: Identification of Interpositional Dependencies Indicating Structurally and/or Functionally Linked Residues
07:08

Optimization of Synthetic Proteins: Identification of Interpositional Dependencies Indicating Structurally and/or Functionally Linked Residues

Published on: July 14, 2015

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
11:27

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

Published on: May 13, 2020

Related Experiment Videos

Last Updated: Jul 12, 2026

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes T&#252;6028
09:08

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028

Published on: January 13, 2017

Optimization of Synthetic Proteins: Identification of Interpositional Dependencies Indicating Structurally and/or Functionally Linked Residues
07:08

Optimization of Synthetic Proteins: Identification of Interpositional Dependencies Indicating Structurally and/or Functionally Linked Residues

Published on: July 14, 2015

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050
11:27

X-Ray Crystallography to Study the Oligomeric State Transition of the Thermotoga maritima M42 Aminopeptidase TmPep1050

Published on: May 13, 2020

Area of Science:

  • Biochemistry
  • Synthetic Biology
  • Natural Product Discovery

Background:

  • Polyketide synthases (PKSs) are crucial for natural product biosynthesis.
  • Acyltransferase (AT) domains dictate extender unit specificity in PKSs.
  • Understanding AT domain sequence-function relationships is key for engineering PKSs.

Purpose of the Study:

  • To identify sequence elements governing extender unit specificity in the EryAT6 acyltransferase domain.
  • To explore the modularity and engineering potential of conserved motifs within AT domains.
  • To develop strategies for accessing diverse polyketides through PKS redesign.

Main Methods:

  • Site-directed mutagenesis of conserved motifs (LSM and SSM) in EryAT6.
  • Functional reconstitution of engineered EryAT6 variants in the Ery6TE system.
  • Analysis of extender unit incorporation using non-native substrates like butylmalonyl-CoA.

Main Results:

  • Specific residue substitutions within LSM and SSM significantly altered extender unit selectivity.
  • A triple-residue mutation enhanced butylmalonyl-CoA incorporation by 42-fold.
  • Exchanging LSM and SSM motifs reprogrammed AT selectivity, enabling the formation of butyl-substituted pyrones.

Conclusions:

  • Compact sequence motifs within AT domains are critical determinants of extender unit specificity.
  • Motif-level engineering of PKS AT domains is a viable strategy for generating novel polyketides.
  • This work provides insights into the rational design of PKS for diverse natural product synthesis.