Characterization of the diastaphenazine/izumiphenazine C biosynthetic gene cluster from plant endophyte Streptomyces diastaticus W2.
Characterization of the diastaphenazine/izumiphenazine C biosynthetic gene cluster from plant endophyte Streptomyces diastaticus W2.
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植物内生菌 Streptomyces diastaticus W2 的二斯塔芬那嗪/izumiphenazine C 生物合成基因簇的表征
DOI：
10.1111/1751-7915.13909
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发表时间：
2022-04
影响因子：
5.7
通讯作者：
He J
中科院分区：
文献类型：
--
作者：
Dong J;He B;Wang R;Zuo X;Zhan R;Hu L;Li Y;He J
从植物内生放线菌链霉菌（Streptomyces diastaticus W2）中分离到两种结构复杂且具有抗肿瘤活性的非那嗪类化合物：地司他那嗪（diastaphenazine）和izumiphenazine C。通过异源表达和基因敲除，鉴定了其推测的生物合成基因簇（dap）。dap基因簇中有20个基因。与莽草通路相关的dap14‐19可能参与前体chorismic酸的生物合成，并且dapBCDEFG被证实负责非那嗪的核结构二苯并吡嗪环的生物合成。两个转录调控基因dapR和dap4在非那嗪生物合成途径中发挥正调控作用。最值得注意的是，环化酶同源基因dap5可以催化地非那嗪中二苯并吡嗪环的二聚化和izumiphenazine C中复杂侧链的负载，这表明一种特殊的修饰策略适合复杂的非那嗪生物合成。此外，基因缺失突变株S. albus::23C5Δdap2的代谢物分析和甲基转移酶Dap2的底物分析清楚地揭示了异源表达和基因失活的方法鉴定了异源表达和基因失活的异源表达和基因失活的方法鉴定了异源非那嗪/异源非那嗪C的生物合成基因簇。通过进一步的功能分析，提出了这两种化合物可能的生物合成途径。值得注意的是，聚酮环化酶Dap5已被证明参与了两个二苯并吡嗪环的二聚化生成地司他非那嗪，以及由Dap2催化甲基化修饰后的复杂侧链的负载生成izumiphenazine C。
Two phenazine compounds, diastaphenazine and izumiphenazine C, with complex structures and promising antitumour activity have been isolated from the plant endophytic actinomycete Streptomyces diastaticus W2. Their putative biosynthetic gene cluster (dap) was identified by heterologous expression and gene knockout. There are twenty genes in the dap cluster. dap14‐19 related to shikimic pathway were potentially involved in the precursor chorismic acid biosynthesis, and dapBCDEFG were confirmed to be responsible for the biosynthesis of the dibenzopyrazine ring, the nuclear structure of phenazines. Two transcriptional regulatory genes dapR and dap4 played the positive regulatory roles on the phenazine biosynthetic pathway. Most notably, the dimerization of the dibenzopyrazine ring in diastaphenazine and the loading of the complex side chain in izumiphenazine C could be catalysed by the cyclase homologous gene dap5, suggesting an unusual modification strategy tailoring complex phenazine biosynthesis. Moreover, metabolite analysis of the gene deletion mutant strain S. albus::23C5Δdap2 and substrate assay of the methyltransferase Dap2 clearly revealed the biosynthetic route of the complex side chain in izumiphenazine C. In this study, we identified the diastaphenazine/izumiphenazine C biosynthetic gene cluster through heterologous expression and gene inactivation. The putative biosynthetic pathway of these two compounds was proposed by further functional analysis. Significantly, the polyketide cyclase Dap5 has been proven to be responsible for the dimerization of two dibenzopyrazine rings to produce diastaphenazine and the loading of the complex side chain after methylation modification catalyzed by Dap2 to generate izumiphenazine C.
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影响因子：
6.8
作者：
Blankenfeldt, Wulf;Parsons, James F.
通讯作者：
Parsons, James F.
影响因子：
3.2
作者：
Lin, Ta-Hui;Wei, Guei-Tsung;Shaw, Gwo-Chyuan
通讯作者：
Shaw, Gwo-Chyuan
影响因子：
4.8
作者：
Li, Qi-Ang;Mavrodi, Dmitri V.;Blankenfeldt, Wulf
通讯作者：
Blankenfeldt, Wulf
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：
5
作者：
Pierson, Leland S., III;Pierson, Elizabeth A.
通讯作者：
Pierson, Elizabeth A.