A one-step method of 10,17-dihydro(pero)xydocosahexa-4Z,7Z,11E,13Z,15E,19Z-enoic acid synthesis by soybean lipoxygenase
A one-step method of 10,17-dihydro(pero)xydocosahexa-4Z,7Z,11E,13Z,15E,19Z-enoic acid synthesis by soybean lipoxygenase
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DOI：
10.1194/jlr.d500042-jlr200
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发表时间：
2006-04-01
影响因子：
6.5
通讯作者：
Butovich, IA
中科院分区：
文献类型：
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作者：
Butovich, IA
二十二碳六烯酸（DHA）的脂氧合酶（LOX）氧化产物，10，17-二氢（佩罗）氧化二十二碳六-4Z，7 Z，11 E，13 Z，15 E，19 Z-烯酸[10，17（S）-二H（P）DHA]通过涉及DHA，17（S）-二氢（佩罗）氧化二十二碳六-4Z，7 Z，11 Z，13 Z，15 E，19 Z-烯酸[ 17（S）-H（P）DHA]、大豆脂氧合酶（sLOX）和马铃薯块茎脂氧合酶（ptLOX）的各种组合。通过HPLC、紫外（UV）光谱、GCMS、串联MS和NMR光谱确认了产物的结构。已经发现，由sLOX通过DHA或17（S）-H（P）DHA的直接氧化形成的10，17（S）-diH（P）DHA与后者的ptLOX氧化产物明显相同。在正相、反相和手性相HPLC分析条件下共洗脱的10，17-diHDHA的sLOX-和ptLOX-衍生样品显示出相同的UV吸收光谱，最大吸收波长为260、270和280 nm，并且具有相似的一维和二维质子NMR光谱。对它们的NMR光谱的分析得出结论，由sLOX形成的10，17-diHDHA仅具有共轭三烯片段的11 E，13 Z，15 E构型，这与先前公布的其ptLOX衍生的对应物的结构相同。根据反应产物的顺、反式几何构型，得出在实验条件下sLOX催化DHA直接双氧化的结论。与先前描述的涉及sLOX和ptLOX的双酶方法相比，当前简化的单酶方法仅使用sLOX作为两个双氧化步骤的催化剂。
A product of lipoxygenase ( LOX) oxidation of docosahexaenoic acid ( DHA), 10,17-dihydro( pero) xydocosahexa-4Z, 7Z, 11E, 13Z, 15E, 19Z-enoic acid [ 10,17( S)-diH( P) DHA] was obtained through various reaction pathways that involved DHA, 17( S)-hydro( pero) xydocosahexa-4Z, 7Z, 11Z, 13Z, 15E, 19Z- enoic acid [ 17( S)-H( P) DHA], soybean lipoxygenase ( sLOX), and potato tuber lipoxygenase ( ptLOX) in various combinations. The structure of the product was confirmed by HPLC, ultraviolet ( UV) light spectrometry, GCMS, tandem MS, and NMR spectroscopy. It has been found that 10,17( S)-diH( P) DHA formed by sLOX through direct oxidation of either DHA or 17( S)-H( P) DHA was apparently identical to the product of ptLOX oxidation of the latter. The sLOX- and ptLOX-derived samples of 10,17-diHDHAs coeluted under the conditions of normal, reverse, and chiral phase HPLC analyses, displayed identical UV absorption spectra with maxima at 260, 270, and 280 nm, and had similar one-dimensional and two-dimensional proton NMR spectra. Analysis of their NMR spectra led to the conclusion that 10,17-diHDHA formed by sLOX had solely 11E, 13Z, 15E configuration of the conjugated triene fragment, which was identical to the previously published structure of its ptLOX-derived counterpart. Based on the cis, trans geometry of the reaction products, the conclusion is made that in the tested conditions sLOX catalyzed direct double dioxygenation of DHA. Compared with the previously described two-enzyme method that involved sLOX and ptLOX, the current simplified one-enzyme procedure uses only sLOX as the catalyst of both dioxygenation steps.