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Cholest 4 en 3 one

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Sourced in China, United States

Cholest-4-en-3-one is a chemical compound used as a laboratory reagent. It is a steroid with a characteristic molecular structure.

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Lab products found in correlation

Cholesterol, by Merck Group (3 mentions) Cholestan 3 one, by Merck Group (1 mentions) Esquire hct ion trap mass spectrometer, by Bruker (1 mentions) 1100 hplc system, by Agilent Technologies (1 mentions) Tween 80, by Merck Group (1 mentions) Tyloxapol, by Merck Group (1 mentions)

4 protocols using cholest 4 en 3 one

1

Analytical Techniques for Sterol Determination

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Cholest-4-en-3-one, cholest-5-en-3-one,
cholesterol, cholestan-3-one, and the solvents and reagents required
for TLC, HPLC, and GC–MS analyses were purchased from Sigma-Aldrich
(Shanghai, China). Cholest-4-en-3-ol and cholest-7-en-3-one were purchased
from Steraloids (Newport, RIUS; https://www.steraloids.com/contact-us), and a methyltransferase colorimetric assay kit was purchased from
NeoBioscience Technology (Shenzhen, China).
Zhou W., Zhang X., Wang A., Yang L., Gan Q., Yi L., Summons R.E., Volkman J.K, & Lu Y. (2022). Widespread Sterol Methyltransferase Participates in the Biosynthesis of Both C4α- and C4β-Methyl Sterols. Journal of the American Chemical Society, 144(20), 9023-9032.
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2

HPLC-MS Analysis of Cholesterol Oxidation

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The total reaction was extracted on 1 mL chloroform. After evaporation of the solvent at room temperature, the product was dissolved in the solvent, which was the same as the mobile phase used for HPLC. 10 μL of the analyte sample were injected into a Phenomenex Gemini® 5 μ C18 110 Å column (250 × 4.6 mm, 5 micron), and chromatography under isocratic conditions was performed using methanol:water 100:2 (v/v) at a flow rate of 0.8 mL/min at room temperature. Cholesterol and cholest-4-en-3-one were purchased from Sigma-Aldrich and used as reference. Product formation was monitored at 200 and 250 nm, whereas cholesterol was detected at 200 nm. The Agilent HPLC 1100 system equipped with a DAD was coupled to an esquireHCT ion trap mass spectrometer (Bruker. Germany), and an atmospheric pressure chemical ionization (APCI) source was operated in the positive ion mode. Conditions were as follows: scan range, m/z 50–600; dry gas flow of 11 L/min, nebulizer pressure 35 psi, drying gas temperature 320°C and the APCI heater temperature was 350°C. The extracted ion current (EIC) signals were deduced based on the exact masses for protonated cholesterol after water elimination (m/z 369.34) as well as for the protonated oxidation product cholest-4-en-3-one (m/z 385.34).
Reiss R., Faccio G., Thöny-Meyer L, & Richter M. (2014). Cloning, expression and biochemical characterization of the cholesterol oxidase CgChoA from Chryseobacterium gleum. BMC Biotechnology, 14, 46.
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3

Cholesterol Extraction Protocol

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Cholesterol, cholest-4-en-3-one, and Tween 80 were purchased from Sigma (USA), and yeast extract was supplied by Oxoid. Horseradish peroxidase was obtained from Sinopharm Chemical Reagent. Inorganic salts and other chemicals were commercial products of analytical grade.
Wu K., Li W., Song J, & Li T. (2015). Production, Purification, and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System. Biochemistry Insights, 8(Suppl 1), 1-8.
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4

Mycobacterium smegmatis Steroid Metabolism

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The strains as well as the plasmids used in this work are listed in Table S1. M. smegmatis mc 2 155 strain was grown at 37°C in an orbital shaker at 250 rpm. Middlebrook 7H9 broth medium (Difco) without albumin-dextrose supplement and without glycerol was used as minimal medium. As carbon source 1.8 mM cholesterol (Sigma), 1.8 mM cholest-4-en-3-one (Sigma), 1 mM phytosterols (Gadea Pharma) or a mixture of 2-10 mM glycerol-1.8 mM cholesterol, all of them dissolved in 3.6% Tyloxapol (Sigma), was added. Due to the low solubility of steroids, stock solutions were warmed at 80ºC with agitation and then autoclaved. As rich medium, 7H9 broth containing 0.2% glycerol, 10% albumin-dextrose supplement and 0.05% Tween 80 or 7H10 agar (Difco) plates supplemented with 0.5% glycerol and 10% albumin-dextrose were used. Gentamycin (5 µg ml -1 ) and kanamycin (25 µg ml -1 ) were used for selection of mycobacteria. Ampicillin (100 µg ml -1 ) and kanamycin (50 µg ml -1 ) were used for plasmid selection and maintenance in Escherichia coli DH10B strain.
García-Fernández J., Papavinasasundaram K., Galán B., Sassetti C.M, & García J.L. (2017). Unravelling the pleiotropic role of the MceG ATPase in Mycobacterium smegmatis. Environmental microbiology, 19(7).
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