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首頁> 美國衛(wèi)生研究院文獻>Biochemistry Insights >Production Purification and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System
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Production Purification and Identification of Cholest-4-en-3-one Produced by Cholesterol Oxidase from Rhodococcus sp. in Aqueous/Organic Biphasic System

機譯：紅球菌屬膽固醇氧化酶產生的Cholest-4-en-3-one的生產純化和鑒定。在水/有機雙相系統(tǒng)中

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摘要

Cholest-4-en-3-one has positive uses against obesity, liver disease, and keratinization. It can be applied in the synthesis of steroid drugs as well. Most related studies are focused on preparation of cholest-4-en-3-one by using whole cells as catalysts, but production of high-quality cholest-4-en-3-one directly from cholesterol oxidase (COD) using an aqueous/organic two-phase system has been rarely explored. This study set up an enzymatic reaction system to produce cholest-4-en-3-one. We developed and optimized the enzymatic reaction system using COD from COX5-6 (a strain of Rhodococcus) instead of whole-cell biocatalyst. This not only simplifies and accelerates the production but also benefits the subsequent separation and purification process. Through extraction, washing, evaporation, column chromatography, and recrystallization, we got cholest-4-en-3-one with purity of 99.78%, which was identified by nuclear magnetic resonance, mass spectroscopy, and infrared spectroscopy. In addition, this optimized process of cholest-4-en-3-one production and purification can be easily scaled up for industrial production, which can largely decrease the cost and guarantee the purity of the product.
機譯：Cholest-4-en-3-one對肥胖，肝病和角質化有積極作用。它也可以用于類固醇藥物的合成。大多數(shù)相關研究都集中在通過使用全細胞作為催化劑來制備cholest-4-en-3-one的方法，但是直接使用膽固醇/水氧化法從膽固醇氧化酶（COD）生產高質量的cholest-4-en-3-one有機兩相系統(tǒng)很少被探索。這項研究建立了一個酶促反應系統(tǒng)以產生cholest-4-en-3-one。我們使用來自COX5-6（紅球菌的一種）的COD代替全細胞生物催化劑，開發(fā)并優(yōu)化了酶促反應系統(tǒng)。這不僅簡化并加快了生產過程，而且有利于后續(xù)的分離和純化過程。通過萃取，洗滌，蒸發(fā)，柱色譜和重結晶，我們獲得了cholest-4-en-3-one，其純度為99.78％，通過核磁共振，質譜和紅外光譜鑒定。另外，這種最優(yōu)化的膽甾4-烯3-一生產和純化方法可以容易地擴大規(guī)模用于工業(yè)生產，這可以大大降低成本并保證產品的純度。

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