Redesign of a dioxygenase in morphine biosynthesis.
Redesign of a dioxygenase in morphine biosynthesis.
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吗啡生物合成中双加氧酶的重新设计。
DOI：
10.1016/j.chembiol.2012.04.017
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发表时间：
2012
影响因子：
--
通讯作者：
Runguphan W
中科院分区：
文献类型：
--
作者：
Runguphan W
罂粟（罂粟）在吗啡喃途径中产生药用重要的苄基异喹啉生物碱，包括镇痛药可待因和吗啡。我们比对了最近在催眠草中发现的三种双加氧酶，并随后确定了非保守区域。已知这些酶中的两种，可待因O-脱甲基酶（PsCODM）和蒂巴因O-脱甲基酶（PsT 6 ODM）促进吗啡喃生物合成中的区域选择性O-脱甲基化。我们系统地交换了PsCODM和PsT 6 ODM序列之间的非保守残基，产生了16个可以在大肠杆菌中过表达的突变体PsCODM蛋白。虽然野生型PsCODM可以使可待因和蒂巴因脱甲基，但一种工程化PsCODM突变体选择性地使可待因脱甲基。在吗啡生物合成的重建中使用这种再工程化的酶可以选择性地使冗余途径分支失活，并因此在随后的代谢工程努力中影响下游产物可待因和吗啡的产率。
Opium poppy (Papaver somniferum) produces medicinally important benzylisoquinoline alkaloids, including the analgesics codeine and morphine, in the morphinan pathway. We aligned three dioxygenases that were recently discovered inP. somniferumand subsequently identified the nonconserved regions. Two of these enzymes, codeine O-demethylase (PsCODM) and thebaine O-demethylase (PsT6ODM), are known to facilitate regioselective O-demethylation in morphinan biosynthesis. We systematically swapped the residues that were nonconserved between thePsCODM andPsT6ODM sequences to generate 16 mutantPsCODM proteins that could be overexpressed inEscherichia coli. While wild-typePsCODM can demethylate both codeine and thebaine, one engineeredPsCODM mutant selectively demethylates codeine. Use of this reengineered enzyme in the reconstitution of morphine biosynthesis could selectively disable a redundant pathway branch and therefore impact the yields of the downstream products codeine and morphine in subsequent metabolic engineering efforts.
印度陆生罂粟 Papaver somniferum L. 的生物碱概况。
影响因子：
2
作者：
S. Prajapati;S. Bajpai;Digvijay Singh;R. Luthra;M. M. Gupta;Sushil Kumar
通讯作者：
Sushil Kumar
DOI：
10.1146/annurev-chembioeng-061010-114209
发表时间：
2011
影响因子：
8.4
作者：
Pickens LB;Tang Y;Chooi YH
通讯作者：
Chooi YH
DOI：
10.1073/pnas.0802981105
发表时间：
2008-05-27
影响因子：
11.1
作者：
Minami, Hiromichi;Kim, Ju-Sung;Sato, Fumihiko
通讯作者：
Sato, Fumihiko