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 Experiment Videos

Multienzyme mevalonate pathway bioreactor.

Autumn Sutherlin1, Victor W Rodwell

  • 1Department of Biochemistry, Purdue University, 175 South University Street, West Lafayette, Indiana 47907-2063, USA.

Biotechnology and Bioengineering
|August 3, 2004
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Targeting Enterococcus faecalis HMG-CoA reductase with a non-statin inhibitor.

Communications biology·2023
Same author

A novel role for coenzyme A during hydride transfer in 3-hydroxy-3-methylglutaryl-coenzyme A reductase.

Biochemistry·2013
Same author

A structural limitation on enzyme activity: the case of HMG-CoA synthase.

Biochemistry·2006
Same author

X-ray crystal structures of HMG-CoA synthase from Enterococcus faecalis and a complex with its second substrate/inhibitor acetoacetyl-CoA.

Biochemistry·2005
Same author

Enterococcus faecalis phosphomevalonate kinase.

Protein science : a publication of the Protein Society·2005
Same author

The 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductases.

Genome biology·2004

Researchers developed a bioreactor assay to screen for new antibiotics targeting the essential mevalonate pathway in Gram-positive bacteria like Enterococcus faecalis.

Area of Science:

  • Biochemistry and Molecular Biology
  • Microbiology
  • Biotechnology

Background:

  • Isopentenyl diphosphate is a fundamental building block for isoprenoid biosynthesis across all life forms.
  • Pathogenic Gram-positive bacteria rely on the mevalonate pathway for survival.
  • Mevalonate pathway enzymes are potential targets for novel antibiotics against multidrug-resistant bacteria.

Purpose of the Study:

  • To develop a rapid, high-throughput bioreactor assay for screening potential inhibitors of the mevalonate pathway.
  • To demonstrate proof of concept using enzymes from the Gram-positive pathogen Enterococcus faecalis.

Main Methods:

  • Immobilization of recombinant enzymes (acetoacetyl-CoA thiolase, HMG-CoA synthase, HMG-CoA reductase) from E. faecalis into two continuous flow bioreactor types: hollow fiber and immobilized plug flow.

Related Experiment Videos

  • Conversion of acetyl-CoA to mevalonate using the immobilized enzymes in both bioreactor systems.
  • Continuous monitoring of reactor performance via spectrophotometric measurement of NADPH concentration.

    • Main Results:

    • Successful immobilization and functional demonstration of key mevalonate pathway enzymes in continuous flow bioreactors.
    • Establishment of a system for monitoring enzyme activity through NADPH consumption.
    • Demonstrated feasibility of using bioreactors for assessing enzyme inhibitors.

    Conclusions:

    • The developed bioreactor assay provides a valuable tool for surveying inhibitors targeting the bacterial mevalonate pathway.
    • This approach can aid in the discovery of new antibiotics against resistant Gram-positive pathogens.
    • Potential for broader applications, including using enzymes from extremophiles for biosynthesis and linking bioreactors to analytical instruments for metabolic studies.