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

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...
Export of Misfolded Proteins out of the ER01:32

Export of Misfolded Proteins out of the ER

After folding, the ER assesses the quality of secretory and membrane proteins. The correctly folded proteins are cleared by the calnexin cycle for transport to their final destination, while misfolded proteins are held back in the ER lumen. The ER chaperones attempt to unfold and refold the misfolded proteins but sometimes fail to achieve the correct native conformation. Such terminally misfolded proteins are then exported to the cytosol by ER-associated degradation or ERAD pathway for...
Production of Pharmaceuticals01:30

Production of Pharmaceuticals

Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
Directing Proteins to the Rough Endoplasmic Reticulum01:34

Directing Proteins to the Rough Endoplasmic Reticulum

The organelle-specific signaling sequences direct proteins synthesized in the cytosol to their final destination like ER, mitochondria, peroxisomes, etc. Some of the proteins directed to ER are then trafficked via vesicles to other organelles within the cell or the extracellular environment through the Golgi complex. For example, the rough ER synthesizes soluble proteins for transportation to the lysosomes or secretion out of the cell. It can also synthesize transmembrane proteins that can...
The Unfolded Protein Response01:37

The Unfolded Protein Response

The ER is the hub of protein synthesis in a cell. It has robust systems to quality control protein folding and also for degradation of terminally misfolded proteins. Under normal conditions, a small proportion of misfolded proteins that cannot be salvaged need to be transported to the cytoplasm by the ER-associated degradation or ERAD pathways. However, if the ERAD cannot handle the misfolded proteins, the cell activates the unfolded protein response or UPR to adjust the protein folding...

You might also read

Related Articles

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

Sort by
Same author

Ribavirin mitigates Alzheimer's disease model phenotypes by enhancing lysosomal function and suppressing ISR.

Bioorganic chemistry·2026
Same author

Biosensor-driven adaptive laboratory evolution and modular pathway engineering for high-level production of nicotinamide mononucleotide in yeast.

Bioresource technology·2026
Same author

Intercropping with Morus alba L. affects Cd accumulation in maize via rhizosphere regulation: Contrasting effects of two Morus alba L. cultivars.

Ecotoxicology and environmental safety·2026
Same author

Engineering Cellobiose Phosphorylase toward Lactose with Its Application to the Biosynthesis of Lacto-<i>N</i>-Biose.

Journal of agricultural and food chemistry·2026
Same author

PDRG1 induced by SP1 facilitates the proliferation and metastasis of hepatocellular carcinoma by activating the Wnt/β-catenin pathway.

MedScience·2026
Same author

<i>Planococcus dechangensis</i> NEAU-ST10-9<sup>T</sup> Promotes Maize Seedling Root Development: Evidence from Effective Fluorescence Tracking.

Microorganisms·2026

Related Experiment Video

Updated: Jul 3, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

Reprogramming the enduracidin NRPS assembly line via thioesterase domain relocation.

Yanan Sun1, Guibin Tu1, Xiaocan Sun1

  • 1Key Laboratory of Industrial Fermentation Microbiology, College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, PR China.

Synthetic and Systems Biotechnology
|July 2, 2026
PubMed
Summary
This summary is machine-generated.

Repositioning a thioesterase (TE) domain in large nonribosomal peptide synthetase (NRPS) assembly lines enables the creation of novel macrocyclic peptides. This engineering strategy allows for programmed backbone lengths and tailored ring sizes in cyclic peptide libraries.

Keywords:
EnduracidinMacrocyclic peptidesNonribosomal peptide synthetase (NRPS)Thioesterase (TE) domain

More Related Videos

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit
08:25

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

Published on: October 31, 2019

Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability
10:31

Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability

Published on: February 3, 2022

Related Experiment Videos

Last Updated: Jul 3, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit
08:25

Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

Published on: October 31, 2019

Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability
10:31

Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability

Published on: February 3, 2022

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Synthetic Biology

Background:

  • Engineering large multi-megadalton nonribosomal peptide synthetase (NRPS) assembly lines is challenging.
  • Module deletion often disrupts macrocyclic peptide formation, limiting analogue generation.

Purpose of the Study:

  • To investigate the potential of thioesterase (TE) domain repositioning for engineering NRPS assembly lines.
  • To generate macrocyclic peptides with programmed backbone lengths by relocating the EndC_TE domain.

Main Methods:

  • Identification of an unusual tri-thioesterase (tri-TE) architecture in the enduracidin NRPS.
  • Systematic relocation of the terminal thioesterase domain (EndC_TE) within the 2.0 MDa enduracidin NRPS.
  • MS/MS analyses to confirm regioselectivity and macrocyclization patterns.

Main Results:

  • Engineered NRPS assembly lines produced macrocyclic peptides with programmed backbone lengths.
  • EndC_TE retained conserved regioselectivity across derivatives, enabling cyclization between specific residues.
  • Successful generation of macrocyclic derivatives with varying lengths.

Conclusions:

  • Thioesterase domain relocation is a viable strategy for reprogramming complex NRPS assembly lines.
  • This approach facilitates the generation of macrocyclic derivatives with tailored ring sizes.
  • EndC_TE is a valuable catalytic domain for constructing cyclic peptide libraries.