Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis.
Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis.
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DOI：
10.1186/s12934-018-0929-4
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发表时间：
2018-05-31
影响因子：
6.4
通讯作者：
Bie X
中科院分区：
文献类型：
--
作者：
Gao L;Guo J;Fan Y;Ma Z;Lu Z;Zhang C;Zhao H;Bie X
普利司他丁是一种抗真菌脂肽，由枯草芽孢杆菌的非核糖体肽合成酶（NRPS）合成。然而，很少有信息的组合生物合成策略应用于plipastatin生物合成途径。在本研究中，我们应用模块或单个结构域删除策略来改造plipastatin生物合成途径，并研究删除对plipastatin装配线的影响，以及揭示新脂肽的合成模式。模块缺失使整个酶复合物失活，而模块7内的单个结构域（A/T结构域）缺失截短了装配线，导致截短的线性六肽（C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ala/瓦尔）。有趣的是，在模块6催化单元内，巯基化结构域缺失的效果不同于腺苷酸化缺失的效果。缺失T6结构域导致非生产性菌株，而缺失A6结构域导致通过模块跳跃机制产生多个装配线，产生三种新型的plipastatin衍生物，五肽（C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu）、六肽（C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ile）和八肽（C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu-Gln-Tyr-Ile）。值得注意的是，在A6结构域删除后发生了独特的模块跳过过程，这在以前的工程NRPS系统中尚未报道过。这一发现为脂肽工程提供了新的见解。这是非常重要的组合方法，并应考虑在工程非核糖体肽生物合成途径，产生新的脂肽。本文的在线版本（10.1186/s12934-018-0929-4）包含补充材料，可供授权用户使用。
Plipastatin, an antifungal lipopeptide, is synthesized by a non-ribosomal peptide synthetase (NRPS) in Bacillus subtilis. However, little information is available on the combinatorial biosynthesis strategies applied in plipastatin biosynthetic pathway. In this study, we applied module or individual domain deletion strategies to engineer the plipastatin biosynthetic pathway, and investigated the effect of deletions on the plipastatin assembly line, as well as revealed the synthetic patterns of novel lipopeptides. Module deletion inactivated the entire enzyme complex, whereas individual domain (A/T domain) deletion within module 7 truncated the assembly line, resulting in truncated linear hexapeptides (C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ala/Val). Interestingly, within the module 6 catalytic unit, the effect of thiolation domain deletion differed from that of adenylation deletion. Absence of the T6-domain resulted in a nonproductive strain, whereas deletion of the A6-domain resulted in multiple assembly lines via module-skipping mechanism, generating three novel types of plipastatin derivatives, pentapeptides (C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu), hexapeptides (C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ile), and octapeptides (C16~17β-OHFA-Glu-Orn-Tyr-Thr-Glu-Gln-Tyr-Ile). Notably, a unique module-skipping process occurred following deletion of the A6-domain, which has not been previously reported for engineered NRPS systems. This finding provides new insight into the lipopeptides engineering. It is of significant importance for combinatorial approaches and should be taken into consideration in engineering non-ribosomal peptide biosynthetic pathways for generating novel lipopeptides. The online version of this article (10.1186/s12934-018-0929-4) contains supplementary material, which is available to authorized users.
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影响因子：
4.2
作者：
Jiang J;Gao L;Bie X;Lu Z;Liu H;Zhang C;Lu F;Zhao H
通讯作者：
Zhao H
影响因子：
5.7
作者：
Samel, Stefan A.;Schoenafinger, Georg;Essen, Lars-Oliver
通讯作者：
Essen, Lars-Oliver
影响因子：
2.9
作者：
Eppelmann, K;Stachelhaus, T;Marahiel, MA
通讯作者：
Marahiel, MA
影响因子：
15
作者：
Mootz, HD;Kessler, N;Marahiel, MA
通讯作者：
Marahiel, MA
影响因子：
5.6
作者：
Samel, Stefan A.;Wagner, Bjoern;Essen, Lars-Oliver
通讯作者：
Essen, Lars-Oliver