Extreme promiscuity of a bacterial and a plant diterpene synthase enables combinatorial biosynthesis.
Extreme promiscuity of a bacterial and a plant diterpene synthase enables combinatorial biosynthesis.
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DOI：
10.1016/j.ymben.2016.04.001
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发表时间：
2016-09
影响因子：
8.4
通讯作者：
Peters RJ
中科院分区：
文献类型：
--
作者：
Jia M;Potter KC;Peters RJ
二萜类化合物广泛分布于许多生物界，它们具有多种生理功能，有些还具有重要的工业用途。它们的生物合成涉及I类二萜合成酶(DTS)，其活性可以先于II类二萜环酶(DTC)。在这里，一个模块化的代谢工程系统被用来检查DTS的混杂。令人惊讶的是，细菌和植物DTS都表现出极端的混杂，与所有可用的前体以正交活性反应，分别产生烯基或羟基。这种DTS的混杂使得组合生物合成成为可能，这些未经优化的非天然酶组合的产量非常高(高达15毫克/L)。事实上，可以很容易地确定这13种未知产品的特征。值得注意的是，其中16个观察到的二萜以前是无法获得的，这些结果提供了生物合成路线，进一步有望使更多延长的途径能够组装成额外精致的“非天然”二萜。
Diterpenes are widely distributed across many biological kingdoms, where they serve a diverse range of physiological functions, and some have significant industrial utility. Their biosynthesis involves class I diterpene synthases (DTSs), whose activity can be preceded by that of class II diterpene cyclases (DTCs). Here, a modular metabolic engineering system was used to examine the promiscuity of DTSs. Strikingly, both a bacterial and plant DTS were found to exhibit extreme promiscuity, reacting with all available precursors with orthogonal activity, producing an olefin or hydroxyl group, respectively. Such DTS promiscuity enables combinatorial biosynthesis, with remarkably high yields for these unoptimized non-native enzymatic combinations (up to 15 mg/L). Indeed, it was possible to readily characterize the 13 unknown products. Notably, 16 of the observed diterpenes were previously inaccessible, and these results provide biosynthetic routes that are further expected to enable assembly of more extended pathways to produce additionally elaborated ‘non-natural’ diterpenoids.