Chiral syntheses by use of substrate specificities of by use of recombinant prenyltransferases

为了研究嗜热脂肪芽孢杆菌法尼基二磷酸合成酶(FPS)的底物专一性，研究了异戊烯基二磷酸(IPP)的4-烷基同系物(IPP)的反应活性。4-甲基同系物(E)-3-甲基戊基-3-烯基二磷酸(1a)和(Z)-3-甲基戊基-3-烯基二磷酸(Ib)分别与香叶基二磷酸(GPP)反应生成4-甲基法呢基二磷酸(2a和2b)。通过圆二色谱确定2a或2b衍生的每一种醛的立体化学分别为(S)-或(R)-4-甲基法呢醛，同样，1a与二甲基烯丙基二磷酸(DMAPP)反应生成(4S)-4-甲基香叶基二磷酸盐(3a)和(4S，8S)-4，8-二甲基法呢基二磷酸(4a)的混合物。4-乙基异构体(E)-3-甲基己基-3-烯基二磷酸(1c)和(Z)-3-甲基己基-3-烯基二磷酸(1D)与…反应得到两种类型的DMAPP更多的4-乙基法呢基二磷酸盐。1C或1D与DMAPP反应还得到两种类型的4-乙基香叶基和4，8-二乙基法呢基二磷酸盐。同时，4-丙基同系物(E)-3-甲基海普特-3-烯基二磷酸(1E)和(Z)-3-甲基海特-3-烯基二磷酸(1f)与GPP反应得到两种类型的4-丙基法呢基二磷酸盐。而(E)-3-甲基-3-烯基二磷酸(1g)或(Z)-3-甲基-3-烯基二磷酸(1h)，或(E)-4-溴-3-甲基-3-烯基二磷酸(1I)或(Z)-4-溴-3-甲基-3-烯基二磷酸(1J)都不能作为高温底物的底物我们利用黄微球菌B-P26的十一烯基二磷酸合成酶(UPS)研究了(E)-3-甲基六-3-烯基二磷酸或(Z)-3-甲基六-3-烯基二磷酸与FPP的酶促反应。(E)-3-甲基己基-3-烯基二磷酸与fpp反应得到手性(4S)-(2Z，6E，10E，14E)-4-ethyl3，7，11，15-tetramethylhexadeca-2，6，10，14-tetraen-1-ol.然而，Z-异构体根本不被接受作为细菌UPS的底物。较少

In order to investigate substrate spcificity of Bacillus stearothermophilus farnesyl diphosphate synthase (FPS), we examined the reactivity of 4-alkyl group homologs of isopentenyl diphosphate (IPP).The enzymatic reactions of the 4-methyl homologs, (E)-3-methylpent-3-enyl diphophate (1a) and (Z)-3methylpent-3-enyl diphophate (Ib) with geranyl diphosphate (GPP) gave 4-methyl-farnesyl diphosphates (2a and 2b), respectively. The stereochemistry of each aldehyde derived from 2a or 2b was determined by CD spectrometry to be (S)- or (R)-4-methylfarnesal, respectively.Similarly, 1a reacted with dimethylallyl diphosphate (DMAPP) to give a mixture of (4S)-4-methylgeranyl diphospahte (3a) and (4S,8S)-4,8-dimethylfarnesyl diphosphate (4a). The Z-isomer 1b also reacted with DMAPP to give the corresponding enantiomers with (4R)- and (4R,8R)-configurations.Reactions of the 4-ethyl homologs, (E)--3-methylhex-3-enyl diphosphate (1c) and (Z)-3-methylhex-3enyl diphosphate (1d) with GPP gave two types of … More 4-ethylfarnesyl diphosphates. Reactions of 1c or 1d with DMAPP also gave two types of 4-ethylgeranyl- and 4,8- diethylfarnesyl diphosphates. Meanwhile, reaction of the 4-propyl homologs, (E)--3-methylhept-3-enyl diphosphate (1e) and (Z)-3-methylhept-3-enyl diphosphate (1f) with GPP gave two types of 4-propylfarnesyl diphosphates. Reactions of 1e or 1f with DMAPP gave only two types of the4-propylGPPsHowever, neither (E)-3-methyloct-3-enyl diphosphate (1g) or (Z)-3-methyloct-3-enyl diphosphate (1h), nor (E)-4-bromo-3-methylbut-3-enyl diphosphate (1i) or (Z)-4-bromo-3-methylbut-3-enyl diphosphate (1j) was acceptable as a substrate for the thermophilic FPS at all.On the other hand, in order to investigate the reactivity of the homologs of homoallylic substrates, we examined the enzymatic reactions of (E)-3-methylhex-3-enyl diphosphate or (Z)-3-methylhex-3-enyl diphosphate with FPP using the undecaprenyl diphosphate synthase (UPS) of Micrococcus luteus B-P 26. (E)-3-methylhex-3-enyl diphosphate reacted with FPP to give a chiral (4S)-(2Z,6E,10E,14E)-4-ethyl3,7,11,15-tetramethylhexadeca-2,6,10,14-tetraen-1-ol. However, the Z-isomer is not accepted as a substrate for the bacterial UPS at all. Less