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

Oligosaccharide Assembly01:24

Oligosaccharide Assembly

Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...
Protein Glycosylation01:25

Protein Glycosylation

Glycosylation, the most common post-translational modification for proteins, serves diverse functions. Adding sugars to proteins makes the proteins more resistant to proteolytic digestion. Glycosylated proteins can act as markers and receptors to promote cell-cell adhesion. Additionally, they have many essential quality control functions in the cell, such as correct protein folding and facilitating transport of misfolded proteins to the cytosol, which can be degraded.
Glycosylation occurs in...
Energy-requiring Steps of Glycolysis01:20

Energy-requiring Steps of Glycolysis

Glucose is the source of nearly all energy used by organisms. The first step of converting glucose into usable energy is called glycolysis. Glycolysis occurs in the cytosol of the cell over two phases: an energy-requiring phase and an energy-releasing phase. Over the first three steps, glucose is converted into different forms and attached to two phosphate groups donated by two ATP molecules, resulting in an unstable sugar. In the next two stages, the unstable sugar splits into two sugar...
Glycolysis: Preparatory Phase01:21

Glycolysis: Preparatory Phase

In cellular metabolism (the complete breakdown of glucose to extract energy),  glycolysis is the first step. Glycolysis takes place in the cytoplasm of both prokaryotic and eukaryotic cells. Glucose enters heterotrophic cells in two ways. One method is through secondary active transport, where the transport takes place against the glucose concentration gradient. The other mechanism uses a group of integral proteins called GLUT proteins, also known as glucose transporter proteins. These...
Bioreactor Controls-III01:22

Bioreactor Controls-III

Strain improvement is a foundational strategy in industrial microbiology aimed at maximizing microbial productivity, particularly because natural isolates typically yield commercially valuable products in very low concentrations. Although optimizing the culture medium and environmental conditions can improve yields, these adjustments are inherently limited by the organism’s genetic potential. As a result, the focus shifts toward genetic modifications to enhance biosynthetic capacity. The...
Protein Folding Quality Check in the RER01:29

Protein Folding Quality Check in the RER

ER is the primary site for the maturation and folding of soluble and transmembrane secretory proteins. The calnexin cycle is a specific chaperone system that folds and assesses the confirmation of N-glycosylated proteins before they can exit the ER lumen. The primary players of this quality check pipeline are the lectins, ER-resident chaperones, and a glucosyl transferase enzyme. In case the calnexin system in the lumen fails to salvage a misfolded protein, it is transported to the cytoplasm...

You might also read

Related Articles

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

Sort by
Same author

Evaluation of vector system in Saccharopolyspora erythraea and construction of new replicative vector.

Applied microbiology and biotechnology·2026
Same author

Rational construction of rpsL or/and rpoB merodiploids affects biosynthesis of secondary metabolite in Streptomyces.

Molecular biology reports·2026
Same author

Genetically engineered Streptomyces viridosporus ATCC 14672 strains for the discovery of novel moenomycins.

Scientific reports·2026
Same author

Genome Mining-Driven Isolation of New Gromomycins and Insights into Their Mode of Action.

ACS chemical biology·2026
Same author

A Hooker Oxygenase Archetype in Polyketide Biosynthesis Challenging the Baeyer-Villiger Monooxygenase Paradigm.

Journal of the American Chemical Society·2026
Same author

Heterologous biosynthetic crosstalk with the native mansouramycin cluster in <i>Streptomyces albus</i> Del14 reveals unexpected metabolites.

RSC chemical biology·2025
Same journal

The Hedgehog Pathway Effector Smoothened Exhibits Signaling Competency in the Absence of Ciliary Accumulation.

Chemistry & biology·2017
Same journal

DIVERSE System: De Novo Creation of Peptide Tags for Non-enzymatic Covalent Labeling by In Vitro Evolution for Protein Imaging Inside Living Cells.

Chemistry & biology·2015
Same journal

Differential Regulation of Specific Sphingolipids in Colon Cancer Cells during Staurosporine-Induced Apoptosis.

Chemistry & biology·2015
Same journal

Synthetic Peptides as cGMP-Independent Activators of cGMP-Dependent Protein Kinase Iα.

Chemistry & biology·2015
Same journal

Unraveling the B. pseudomallei Heptokinase WcbL: From Structure to Drug Discovery.

Chemistry & biology·2015
Same journal

Vitamin C as Cancer Destroyer, Investigating Sulfhydration, and the Variability in CFTR Interactome.

Chemistry & biology·2015
See all related articles

Related Experiment Video

Updated: Jun 26, 2026

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells
06:53

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells

Published on: June 2, 2022

Engineering a function into a glycosyltransferase.

Christine Krauth1, Marta Fedoryshyn, Christian Schleberger

  • 1Albert-Ludwigs-Universität, Institut für Pharmazeutische Wissenschaften, Freiburg, Germany.

Chemistry & Biology
|January 28, 2009
PubMed
Summary
This summary is machine-generated.

Engineering glycosyltransferases is crucial due to their specific substrate recognition. Researchers introduced new activity into a landomycin E glycosyltransferase by identifying key amino acids that dictate substrate specificity.

More Related Videos

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility
12:29

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

Published on: March 11, 2022

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles
14:37

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

Published on: July 6, 2012

Related Experiment Videos

Last Updated: Jun 26, 2026

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells
06:53

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells

Published on: June 2, 2022

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility
12:29

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

Published on: March 11, 2022

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles
14:37

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

Published on: July 6, 2012

Area of Science:

  • Biochemistry and Molecular Biology
  • Enzymology
  • Natural Product Biosynthesis

Background:

  • Glycosyltransferases (GTs) are enzymes that catalyze glycosylation, a critical step in modifying natural products.
  • Natural GTs often exhibit narrow substrate specificity, limiting their biotechnological applications.
  • Engineering GTs to alter or expand substrate specificity is vital for synthetic biology and drug discovery.

Purpose of the Study:

  • To engineer a glycosyltransferase involved in landomycin E biosynthesis to exhibit novel activity.
  • To identify and leverage 'hot spot' amino acid residues that control GT substrate specificity.

Main Methods:

  • Bioinformatic analysis to identify potential hot spot amino acids in glycosyltransferases.
  • Site-directed mutagenesis of the landomycin E glycosyltransferase based on identified hot spots.
  • Enzymatic assays to evaluate the substrate specificity of engineered variants.

Main Results:

  • Successfully introduced a new enzymatic activity into the target glycosyltransferase.
  • Identified specific amino acid residues critical for determining substrate specificity.
  • Demonstrated the feasibility of engineering GTs by targeting key residues.

Conclusions:

  • Targeted engineering of glycosyltransferases by focusing on substrate-determining hot spot amino acids is an effective strategy.
  • This approach can expand the utility of natural product biosynthetic enzymes for biotechnological applications.
  • The engineered landomycin E glycosyltransferase offers potential for novel glycosylation reactions.