Process for production of 5-ene-3-one or 3,6-dione...

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing a...

Reexamination Certificate

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Reexamination Certificate

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08008040

ABSTRACT:
An object of the present invention is to provide a process according to which 5-ene-3-one or 3,6-dione derivatives of sterols can be synthesized with a better yield, as well as a process for production of lipid metabolism improvers, foods, drinks, and animal feeds containing the 5-ene-3-one or 3,6-dione derivatives of sterols.The process for production of 5-ene-3-one or 3,6-dione derivatives of sterols of the present invention involves preparing a culturing solution containing anArthrobacterbacterium, which is a microorganism exhibiting a cholesterol oxidase activity, and then, adding to the culturing solution an equivalent amount of hexane so as to prepare a bilayer solution consisting of an aqueous layer and a hydrocarbon-based solvent layer. Thereafter, sterol, or a derivative thereof, is made as a substrate contained therein, a reaction performed with cholesterol oxidase at 30° C. for 2 days, and, ethanol then added to separate a hydrocarbon-based solvent layer. Thereafter, the layer is dried to a state of dryness by means of a vacuum dryer so as to obtain 5-ene-3-one or 3,6-dione derivatives.

REFERENCES:
patent: 2003/0153051 (2003-08-01), Aono et al.
patent: H11-193296 (1999-07-01), None
patent: 2001-055333 (2001-02-01), None
patent: 2001-240544 (2001-09-01), None
patent: WO 99/45106 (1999-09-01), None
Dai Kitamoto. “Reaction mechanism of cholesterol oxidase”, Bioscience and Industry, vol. 57, No. 7, 1999, pp. 464-467.
Liu, et al. “Bioconversion of cholesterol to cholest-4-en-3-one in aqueous/organic solvent two-phase reactors”, Enzyme and Microbial Technology, vol. 18, 1996, pp. 184-189.
Michihisa, et al. “Screening and application of organic solvent resistant-microorganisms”, Bioscience and Industry, vol. 61, No. 1, 2003, pp. 17-21.
Yamashita, et al. “Separation of the two reactions, oxidation and isomerization, catalyzed byStreptomycescholesterol oxidase”, Protein Engineering, vol. 11, No. 11,1998, pp. 1075-1079.
Doukyu, et al. “Control with organic solvents of efficiency of persolvent cholesterol fermentation byPseudomonassp. strain ST-200”, Biosci. Biotech. Biochem, vol. 60, No. 10, 1996, pp. 1612-1616.
Aono, R. et al.,Oxidative Bioconversion of Cholesterol by Pseudomonas sp. Strain ST-200 in a Water-Organic Solvent Two-Phase System; Applied and Environmental Microbiology, vol. 60, No. 7, Jul. 1994, pp. 2518-2523.
Dong (1988) Separation of Mixed Sterol by High Performance Liquid Chromatography, Cereals & Oils, 3:40-41.
Razzazi-Fazeli et al. (2000) Determination of Cholesterol Oxides in Processed Food Using High-Performance Liquid Chromatography-Mass Spectrometry With Atmospheric Pressure Chemical Ionisation, J. Chromatogr. A., 896:321-334.

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