Functional and Structural Analysis of Phenazine O-Methyltransferase LaPhzM from Lysobacter antibioticus OH13 and One-Pot Enzymatic Synthesis of the Antibiotic Myxin.
Functional and Structural Analysis of Phenazine O-Methyltransferase LaPhzM from Lysobacter antibioticus OH13 and One-Pot Enzymatic Synthesis of the Antibiotic Myxin.
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DOI：
10.1021/acschembio.8b00062
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发表时间：
2018-04-20
影响因子：
4
通讯作者：
Du L
中科院分区：
文献类型：
--
作者：
Jiang J;Guiza Beltran D;Schacht A;Wright S;Zhang L;Du L
Myxin是一种众所周知的抗生素，已经使用了几十年。它属于吩嗪类天然产物，具有多种生物活性，这些活性通常由杂芳三环系统上的装饰基团决定。粘菌素的三个环带有许多装饰物，包括一种不寻常的芳香N5，N10-二氧化物。我们以前的研究表明，吩嗪1，6-二羧酸（PDC）是粘菌素的直接前体，两种氧化还原酶（LaPhzS和LaPhzNO 1）催化PDC的脱羧羟基化和芳香族N-氧化产生碘（1.6-二羟基-N5，N10-二氧化吩嗪）。在这项工作中，我们确定了LaPhzM基因溶杆菌OH 13，并证明LaPhzM编码SAM依赖的O-甲基转移酶转化碘到粘菌素。结果进一步表明，LaPhzM负责从菌株OH 13分离的所有吩嗪化合物中的单甲氧基和二甲氧基形成。LaPhzM表现出宽松的底物选择性，催化吩嗪与非-，单-或二-N-氧化物的O-甲基化。此外，我们证明了一锅生物合成的粘菌素在体外重建的三个吩嗪环装饰酶。最后，我们确定了X射线晶体结构的LaPhzM与一个绑定的辅因子在1.4 <$m分辨率。该结构提供了第一个特征吩嗪O-甲基转移酶的活性和选择性的分子见解。这些结果将促进未来开发的吩嗪类药物作为新的抗生素，通过代谢工程和化学酶合成。
Myxin is a well-known antibiotic that had been used for decades. It belongs to the phenazine natural products that exhibit various biological activities, which are often dictated by the decorating groups on the heteroaromatic three-ring system. The three rings of myxin carry a number of decorations, including an unusual aromatic N5,N10-dioxide. We previously showed that phenazine 1,6-dicarboxylic acid (PDC) is the direct precursor of myxin, and two redox enzymes (LaPhzS and LaPhzNO1) catalyze the decarboxylative hydroxylation and aromatic N-oxidations of PDC to produce iodinin (1.6-dihydroxy-N5,N10-dioxide phenazine). In this work, we identified the LaPhzM gene from Lysobacter antibioticus OH13 and demonstrated that LaPhzM encodes a SAM-dependent O-methyltransferase converting iodinin to myxin. The results further showed that LaPhzM is responsible for both monomethoxy and dimethoxy formation in all phenazine compounds isolated from strain OH13. LaPhzM exhibits relaxed substrate selectivity, catalyzing O-methylation of phenazines with non-, mono-, or di-N-oxide. In addition, we demonstrated a one-pot biosynthesis of myxin by in vitro reconstitution of the three phenazine-ring decorating enzymes. Finally, we determined the X-ray crystal structure of LaPhzM with a bound cofactor at 1.4 Å resolution. The structure provided molecular insights into the activity and selectivity of the first characterized phenazine O-methyltransferase. These results will facilitate future exploitation of the thousands of phenazines as new antibiotics through metabolic engineering and chemoenzymatic syntheses.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
3.6
作者：
Dietrich, Lars E. P.;Price-Whelan, Alexa;Newman, Dianne K.
通讯作者：
Newman, Dianne K.
影响因子：
4.4
作者：
Mavrodi, Dmitri V.;Peever, Tobin L.;Thomashow, Linda S.
通讯作者：
Thomashow, Linda S.
影响因子：
3.6
作者：
Glasser NR;Kern SE;Newman DK
通讯作者：
Newman DK
影响因子：
14.9
作者：
Davis IW;Leaver-Fay A;Chen VB;Block JN;Kapral GJ;Wang X;Murray LW;Arendall WB 3rd;Snoeyink J;Richardson JS;Richardson DC
通讯作者：
Richardson DC