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

Acinetobacter cyclohexanone monooxygenase: gene cloning and sequence determination.

Y C Chen1, O P Peoples, C T Walsh

  • 1Department of Biology and Chemistry, Massachusetts Institute of Technology, Cambridge 02139.

Journal of Bacteriology
|February 1, 1988
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

Structural characterization of enterobactin hydrolase IroE.

Biochemistry·2006
Same author

NovJ/NovK catalyze benzylic oxidation of a beta-hydroxyl tyrosyl-S-pantetheinyl enzyme during aminocoumarin ring formation in novobiocin biosynthesis.

Biochemistry·2005
Same author

Natural product biosynthetic assembly lines: prospects and challenges for reprogramming.

Ernst Schering Research Foundation workshop·2005
Same author

Antibiotic glycosyltransferases.

Biochemical Society transactions·2003
Same author

Chain termination steps in nonribosomal peptide synthetase assembly lines: directed acyl-S-enzyme breakdown in antibiotic and siderophore biosynthesis.

Chembiochem : a European journal of chemical biology·2002
Same author

Glycopeptide antibiotic biosynthesis: enzymatic assembly of the dedicated amino acid monomer (S)-3,5-dihydroxyphenylglycine.

Proceedings of the National Academy of Sciences of the United States of America·2001
Same journal

The bacterial SOS response promotes the expression of the transposase encoded by IS<i>CR</i> mobile genetic elements.

Journal of bacteriology·2026
Same journal

Development of a gene-editing strategy to overcome genetic intractability in <i>Lactobacillus johnsonii</i>.

Journal of bacteriology·2026
Same journal

Bactofilins are essential spatial organizers of peptidoglycan insertion in the Lyme disease spirochete <i>Borrelia burgdorferi</i>.

Journal of bacteriology·2026
Same journal

DNA damage-associated vesicle production in <i>Stenotrophomonas maltophilia</i> is mediated by the maltocin endolysin.

Journal of bacteriology·2026
Same journal

Characterization of <i>Helicobacter pylori</i> aggregation reveals a requirement for both AlpA and AlpB.

Journal of bacteriology·2026
Same journal

Cross-regulation of amino acid synthesis and anaerobic electron transfer by MetR-mediated methionine signaling.

Journal of bacteriology·2026
See all related articles

Researchers isolated and sequenced the cyclohexanone monooxygenase gene from Acinetobacter sp. strain NCIB 9871. They identified key enzyme binding sites and developed an overproduction system for genetic studies.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Cyclohexanone monooxygenase (CHMO) is an important enzyme in various metabolic pathways.
  • Understanding the genetic basis of CHMO is crucial for its biotechnological applications.

Purpose of the Study:

  • To isolate and characterize the gene encoding cyclohexanone monooxygenase from Acinetobacter sp. strain NCIB 9871.
  • To determine the nucleotide and amino acid sequences of the CHMO gene and its product.
  • To identify potential flavin- and nicotinamide-binding sites within the enzyme and construct an overproduction system.

Main Methods:

  • Immunological screening methods were employed to isolate the CHMO gene.
  • Nucleotide sequencing was performed to determine the gene's structure and flanking regions.

Related Experiment Videos

  • Bioinformatic analysis was used to derive the amino acid sequence and identify conserved domains.
  • An overproduction system was constructed using the pKK223-3 vector in Escherichia coli.

    • Main Results:

    • The structural gene for CHMO was isolated and sequenced, comprising 1,626 nucleotides.
    • The gene codes for a polypeptide of 542 amino acids, with 389 nucleotides upstream and 108 nucleotides downstream of the coding region reported.
    • Potential flavin-binding (residues 6-18) and nicotinamide-binding (residues 176-208) sites were identified.
    • A functional overproduction system for the CHMO gene was successfully established in E. coli.

    Conclusions:

    • The nucleotide and amino acid sequences of Acinetobacter sp. CHMO provide insights into its structure and function.
    • Identification of binding sites aids in understanding the enzyme's catalytic mechanism.
    • The developed overproduction system facilitates further genetic manipulation and protein engineering of CHMO.