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

Products of the Citric Acid Cycle00:53

Products of the Citric Acid Cycle

The cells of most organisms—including plants and animals—obtain usable energy through aerobic respiration, the oxygen-requiring version of cellular respiration. Aerobic respiration consists of four major stages: glycolysis, pyruvate oxidation, the citric acid cycle, and oxidative phosphorylation. The third major stage, the citric acid cycle, is also known as the Krebs cycle or tricarboxylic acid (TCA) cycle.
Amino Acid Biosynthetic Pathways01:29

Amino Acid Biosynthetic Pathways

Amino acid biosynthesis is essential for cell growth, protein synthesis, and metabolic regulation. Cells generate essential and non-essential amino acids from metabolic intermediates to sustain vital biological functions. These intermediates originate from key metabolic pathways: glycolysis, the tricarboxylic acid (TCA) cycle, and the pentose phosphate pathway. Important precursors include α-ketoglutarate, pyruvate, oxaloacetate, phosphoenolpyruvate, and erythrose-4-phosphate, which provide...
Biosynthesis of Nucleic Acids01:28

Biosynthesis of Nucleic Acids

Nucleic acid biosynthesis is a fundamental biochemical process that produces the purine and pyrimidine nucleotides essential for DNA and RNA synthesis. This pathway maintains a balanced nucleotide pool, preventing imbalances that could jeopardize genetic integrity and cellular function. Given the crucial role of nucleotides, their synthesis is tightly regulated to ensure proper cellular homeostasis.Purine BiosynthesisThe biosynthesis of purine nucleotides begins with ribose-5-phosphate, a...
Biosynthesis of Lipids01:29

Biosynthesis of Lipids

Microbial membranes exhibit remarkable diversity in lipid composition, reflecting evolutionary adaptations to various environmental conditions. The three domains of life—Bacteria, Archaea, and Eukarya—synthesize membrane lipids through distinct biosynthetic pathways, leading to fundamental structural differences that impact membrane stability, function, and adaptability.Fatty Acid-Based Lipids in Bacteria and EukaryaBacteria and eukaryotes share a common fatty acid biosynthesis pathway, which...
Production of Antibiotics01:27

Production of Antibiotics

Penicillin, one of the earliest and most widely used antibiotics, is produced industrially by the filamentous fungus Penicillium chrysogenum. Large stirred-tank bioreactors ranging from tens to hundreds of thousands of liters maintain tightly controlled temperature, pH, and dissolved oxygen conditions to support fungal metabolism and maximize antibiotic yield. Penicillin is a secondary metabolite, synthesized primarily during the stationary growth phase, which requires a carefully managed...
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...

You might also read

Related Articles

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

Sort by
Same author

Metabolic engineering of Rhodotorula toruloides IFO0880 improves C16 and C18 fatty alcohol production from synthetic media.

Microbial cell factories·2022
Same author

Selectively Lighting Up Singlet Oxygen via Aggregation-Induced Electrochemiluminescence Energy Transfer.

Analytical chemistry·2022
Same author

Directed Evolution of Replication-Competent Double-Stranded DNA Bacteriophage toward New Host Specificity.

ACS synthetic biology·2022
Same author

Metabolic engineering of threonine catabolism enables Saccharomyces cerevisiae to produce propionate under aerobic conditions.

Biotechnology journal·2022
Same author

Recent advances and perspectives of enzyme-based optical biosensing for organophosphorus pesticides detection.

Talanta·2021
Same author

Pathological diagnosis, differential diagnosis and origin investigation of easily misdiagnosed adult gastric duplication cysts.

Histology and histopathology·2021

Related Experiment Video

Updated: Jul 18, 2026

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028
09:08

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

Published on: January 13, 2017

Biosynthesis of phloroglucinol.

Jihane Achkar1, Mo Xian, Huimin Zhao

  • 1Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.

Journal of the American Chemical Society
|April 14, 2005
PubMed
Summary

Pseudomonas fluorescens Pf-5 synthesizes phloroglucinol using the phlACBDE gene cluster. The enzyme PhlD, derived from this cluster, efficiently converts malonyl-CoA into phloroglucinol, enabling sustainable production.

Area of Science:

  • Biochemistry
  • Microbial Biotechnology
  • Enzyme Catalysis

Background:

  • Phloroglucinol and its derivatives are valuable compounds with diverse applications.
  • Biosynthesis pathways for phloroglucinol derivatives are complex and often involve multiple enzymes.
  • Understanding microbial synthesis offers potential for sustainable production methods.

Purpose of the Study:

  • To identify and characterize the enzyme responsible for phloroglucinol biosynthesis in Pseudomonas fluorescens Pf-5.
  • To investigate the enzymatic conversion of malonyl-CoA to phloroglucinol.
  • To explore the potential for bio-based synthesis of phloroglucinol and related compounds.

Main Methods:

  • Gene expression of the phlACBDE cluster and individual genes (phlD) in Escherichia coli.

More Related Videos

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade
09:50

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade

Published on: August 14, 2019

A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
07:59

A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products

Published on: October 4, 2019

Related Experiment Videos

Last Updated: Jul 18, 2026

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028
09:08

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

Published on: January 13, 2017

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade
09:50

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade

Published on: August 14, 2019

A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
07:59

A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products

Published on: October 4, 2019

  • Purification of the PhlD enzyme to homogeneity.
  • Enzymatic assays to determine kinetic parameters (Km, kcat) for malonyl-CoA conversion.
  • Analysis of acetylase and deacetylase activities of related enzymes.

    • Main Results:

    • Expression of the phlACBDE cluster in E. coli resulted in substantial phloroglucinol synthesis.
    • Expression of the single gene phlD led to extracellular accumulation of phloroglucinol.
    • Purified PhlD enzyme efficiently catalyzed the conversion of malonyl-CoA to phloroglucinol (Km = 5.6 μM, kcat = 10 min⁻¹).
    • Enzymes from the phlACB genes exhibited acetylase and deacetylase activities, interconverting phloroglucinol and its acetylated forms.

    Conclusions:

    • PhlD is identified as the key enzyme catalyzing the formation of phloroglucinol from malonyl-CoA.
    • This discovery provides mechanistic insights into phloroglucinol biosynthesis.
    • PhlD offers a foundation for developing environmentally friendly methods for synthesizing phloroglucinol and resorcinol from glucose.