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

Design Example: Application of Archimedes' Principle01:11

Design Example: Application of Archimedes' Principle

870
Archimedes' principle is fundamental in analyzing the buoyant force and stability of floating bodies. In this example, a wooden block with a rectangular section floats in seawater. Based on the block's dimensions, its specific gravity and the specific weight of seawater are used to find the volume of water displaced and the center of buoyancy.
The volume of seawater displaced by the block is determined by first calculating the block's weight. This is done by multiplying the...
870
Rationalizing Substitutions01:29

Rationalizing Substitutions

62
Integrals involving non-rational functions are often difficult to evaluate using standard techniques, especially when radicals appear in the integrand. Rationalizing substitution provides a systematic method for simplifying such integrals by converting them into rational forms that are easier to handle.Consider a rod whose linear mass density depends on a constant linear density, a characteristic length, and the distance from the left end of the rod. Determining the total mass requires...
62
Rational Expressions01:28

Rational Expressions

407
Rational expressions are algebraic fractions in which both the numerator and the denominator are polynomials. These expressions follow the arithmetic rules of numerical fractions but require extra care due to the presence of variables. A fundamental part of working with rational expressions is identifying values that make the expression undefined, typically those that result in division by zero or undefined radicals.Determining the DomainThe domain of a rational expression includes all real...
407
What is Metabolism?00:52

What is Metabolism?

132.0K
Overview
132.0K
Asymptotes in Rational Functions01:30

Asymptotes in Rational Functions

262
A rational function is defined as the quotient of two polynomials:  where Q(x)≠0, These functions often exhibit asymptotes, which are the lines that the graph approaches but never touches. These asymptotes are classified based on how the function behaves near specific values of the input.Vertical asymptotes occur where the denominator is zero, and the numerator is not, causing the function to be undefined. These are found by solving Q(x)=0. For example:  has a vertical...
262
Factorial Design02:01

Factorial Design

14.0K
Factorial Analysis is an experimental design that applies Analysis of Variance (ANOVA) statistical procedures to examine a change in a dependent variable due to more than one independent variable, also known as factors. Changes in worker productivity can be reasoned, for example, to be influenced by salary and other conditions, such as skill level. One way to test this hypothesis is by categorizing salary into three levels (low, moderate, and high) and skills sets into two levels (entry level...
14.0K

You might also read

Related Articles

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

Sort by
Same author

Biodegradability of Acrylate-Lipoic Acid Copolymers.

Journal of the American Chemical Society·2026
Same author

Engineered Gram-Positive Based Quorum Sensing for Metabolic Control in <i>Escherichia coli</i>.

ACS synthetic biology·2025
Same author

Deciphering allosterism of an <i>Escherichia coli</i> hexuronate metabolism regulator: UxuR.

RSC medicinal chemistry·2025
Same author

Substrate Turnover Dynamics Guide Ketol-Acid Reductoisomerase Redesign for Increased Specific Activity.

ACS catalysis·2024
Same author

Author Correction: A genomic mutational constraint map using variation in 76,156 human genomes.

Nature·2024
Same author

A genomic mutational constraint map using variation in 76,156 human genomes.

Nature·2023

Related Experiment Video

Updated: Feb 9, 2026

Mouse Genome Engineering Using Designer Nucleases
12:04

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

29.3K

Rational design of thiolase substrate specificity for metabolic engineering applications.

Brian M Bonk1,2, Yekaterina Tarasova3, Michael A Hicks4

  • 1Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts.

Biotechnology and Bioengineering
|June 8, 2018
PubMed
Summary
This summary is machine-generated.

Metabolic engineering of the 3-hydroxyacid (3HA) pathway was improved by rationally designing thiolase enzymes. This resulted in a nearly 10-fold increase in the production of desired longer-chain compounds, enhancing efficiency.

Keywords:
metabolic engineeringprotein engineeringthiolase

More Related Videos

Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

15.6K
Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications
05:20

Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications

Published on: May 31, 2018

15.4K

Related Experiment Videos

Last Updated: Feb 9, 2026

Mouse Genome Engineering Using Designer Nucleases
12:04

Mouse Genome Engineering Using Designer Nucleases

Published on: April 2, 2014

29.3K
Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

15.6K
Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications
05:20

Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications

Published on: May 31, 2018

15.4K

Area of Science:

  • Biocatalysis and Metabolic Engineering
  • Enzyme Engineering
  • Synthetic Biology

Background:

  • Metabolic engineering requires highly active and specific enzymes for commercial viability.
  • The 3-hydroxyacid (3HA) pathway synthesizes valuable compounds but suffers from byproduct formation, reducing yields.
  • Thiolase enzymes initiate the 3HA pathway, and their substrate specificity dictates product chain length.

Purpose of the Study:

  • To rationally engineer thiolase substrate specificity to enhance the production of longer-chain products in the 3HA pathway.
  • To increase the ratio of C6/C4 products (3-hydroxy-hexanoic acid/3-hydroxybutyric acid) synthesized via the 3HA pathway.

Main Methods:

  • Employed a model-guided, rational design approach focusing on ordered substrate binding.
  • Applied this design strategy to two specific biosynthetic thiolases.
  • Screened and identified thiolase mutants with altered substrate specificity.

Main Results:

  • Identified specific thiolase mutants that significantly enhance C6/C4 product selectivity.
  • Achieved nearly 10-fold increases in the ratio of C6/C4 products formed.
  • Demonstrated the effectiveness of rational design in altering enzyme specificity for metabolic engineering.

Conclusions:

  • Rational engineering of thiolase substrate specificity can overcome byproduct formation in the 3HA pathway.
  • The developed strategy can improve yields of desired longer-chain products, reducing downstream costs.
  • Findings contribute to understanding enzyme sequence-structure-function relationships and can be applied to other chain-elongation pathways.