Separation and partial characterization of soluble fatty acid alpha-hydroxylase from Sphingomonas paucimobilus.
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.
Matsunaga
I
I
Toneyama Institute for Tuberculosis Research, Osaka City University Medical School, Japan.
Kusunose
E
E
Yano
I
I
Ichihara
K
K
D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins; D002474:Cell-Free System; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification*; D006899:Mixed Function Oxygenases / Q000302:isolation & purification*; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D009251:NADPH-Ferrihemoprotein Reductase; D010100:Oxygen / Q000378:metabolism; D012995:Solubility
D001426:Bacterial Proteins; D009243:NAD; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases; D010100:Oxygen