Matsunaga I, Kusunose E, Yano I, Ichihara K
Toneyama Institute for Tuberculosis Research, Osaka City University Medical School, Japan.
Biochem Biophys Res Commun. 1994 Jun 30;201(3):1554-60. doi: 10.1006/bbrc.1994.1881.
The crude extracts from Sphingomonas paucimobilus containing 2-hydroxy fatty acid-rich sphingolipids were found to oxidize [1-14C]-myristate to 2-hydroxymyristate in the presence of NADH. The myristate-oxidation activity was partially purified about 290-fold from the cell-free extracts by ammonium sulfate fractionation and hydroxylapatite chromatography. When laurate, myristate, and palmitate were used as the substrates, the reaction products were identified as the corresponding 2-hydroxy fatty acids by gas-chromatography (GC), HPLC, and GC-mass spectrometry. The enzyme preparation required NADH and molecular oxygen for its alpha-hydroxylation activity, suggesting that the bacterial alpha-hydroxylation is typical of monooxygenase reactions. Of fatty acids tested, myristate was the most efficient substrate.
研究发现，从少动鞘氨醇单胞菌中提取的含有富含2-羟基脂肪酸鞘脂的粗提物，在NADH存在的情况下能将[1-14C]-肉豆蔻酸氧化为2-羟基肉豆蔻酸。通过硫酸铵分级分离和羟基磷灰石色谱法，从无细胞提取物中对肉豆蔻酸氧化活性进行了部分纯化，纯化倍数约为290倍。当使用月桂酸、肉豆蔻酸和棕榈酸作为底物时，通过气相色谱（GC）、高效液相色谱（HPLC）和GC-质谱法将反应产物鉴定为相应的2-羟基脂肪酸。该酶制剂的α-羟基化活性需要NADH和分子氧，这表明细菌的α-羟基化是典型的单加氧酶反应。在所测试的脂肪酸中，肉豆蔻酸是最有效的底物。
