Bioconversion and Identification of Phytostenones from Phytosterol by a Mutant of Arthrobacter simplex

利用關節桿菌變異株將植物固醇轉換成植物固酮及其鑑定研究

黃承烺(Chen-Lang Huang) ； 劉文雄(Wen-Hsiung Liu)

《臺灣農業化學與食品科學》 44卷3期 (2006/06) Pp. 157-166

https://doi.org/10.6578/TJACFS.2006.018

摘要

本研究利用誘導性膽固醇氧化酶生產變異株Arthrobacter simplex USA18在每公升含有10克植物固醇之培養基中將植物固醇(phytosterol)轉換成植物固酮(Phytostenones)。在5公升醱酵槽中培養、120小時後，可獲得植物固酮之最高蓄積量(8.5mM)，其轉換率為38.5%。利用配備MetaChem Polaris C18-A管柱(5μm, 500×4.6mm)之高效液相層析儀，在移動相為100%甲醇、流速為1.2mL/min之條件下，可有效地分離三種主要植物固酮。三種主要植物固酮經結晶、熔點測定、紅外線光譜、紫外線及可見光光譜、核磁共振光譜、質譜分析等物理及儀器分析後，鑑定其結構分別為菜籽固酮(campest-4-en-3-one;Ⅱ)、豆固酮(stigmasta-4,22-dien-3-one;Ⅰ)及穀固酮(sitost-4-en-3-one;Ⅲ)。

關鍵字

植物固酮 ； 植物固醇 ； 菜籽固酮 ； 豆固酮 ； 穀固酮 ； 膽固醇氧化酶

並列摘要

In this study phytosterol was converted to phytostenones by an inducible cholesterol oxidase-producing mutant, Arthrobacter simplex USA18, in a medium containing phytosterol 10 g, per liter of distilled water. A maximum accumulation of phytostenones (8.5 mM) was obtained after 120 h of cultivation in a 5 L fermentor, and the conversion rate was 38.5%. To separate the mixture of the three major phytostenones from the fermentation broth, a high performance liquid chromatography (HPLC) method was carried out under the following conditions, column: MetaChem Polaris C18-A (5μm particles, 500×4.6 mm I.D.), mobile phase: 100% methanol, and flow rate: 1.2 mL/min. According to the physical and instrumental analyses including crystallization, melting point, ESI-MS, IR, UV-Vis and 1H-NMR spectrometries, the three major phytostenones present in the fermentation broth were campest-4-en-3-one (Ⅱ), stigmasta-4,22-dien-3-one (Ⅰ) and sitost-4-en-3-one (Ⅲ), respectively.

並列關鍵字

Phytostenone ； Phytosterol ； Campestenone ； Stigmastadienone ； Sitostenone ； Cholesterol oxidase