Combinatorial assembly platform enabling engineering of genetically stable metabolic pathways in cyanobacteria.
Combinatorial assembly platform enabling engineering of genetically stable metabolic pathways in cyanobacteria.
复制标题
组合组装平台，使蓝藻遗传稳定代谢途径的工程。
DOI：
10.1093/nar/gkab791
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发表时间：
2021-12-02
影响因子：
14.9
通讯作者：
Heap JT
中科院分区：
文献类型：
--
作者：
Taylor GM;Hitchcock A;Heap JT
蓝藻是简单、高效、基因易驯化的光合作用微生物，原则上可以代表二氧化碳捕获和转化的理想生物催化剂。然而，在实践中，遗传不稳定和低生产率是工程蓝藻的关键、相关问题。我们采取了大规模平行的方法，为蓝藻聚球藻sp.生成并鉴定了合成启动子和RBSS的文库。PCC 6803，并组装了一个稀疏的组合库，该文库包含数百万个编码代谢途径的构造变体。观察到一些变异的遗传不稳定，当变异造成代谢负担时，这是预期的。然而，令人惊讶的是，在没有迭代优化的一轮组合中，随机选择的80%的变异体在经过多代光自养培养后，从大气二氧化碳中积累了目标萜类番茄红素，显然克服了遗传不稳定性。这种大规模的蓝藻并行代谢工程为开发遗传稳定的蓝藻生物催化剂提供了一个新的平台，用于直接从二氧化碳中可持续地利用光驱动生产有价值的产品，避免了化石碳或与食品生产的竞争。从具有合适性质的合成部分中组合组装代谢途径编码库，然后进行少量筛选，是获得蓝藻聚胞藻高产且遗传稳定的途径设计的一种快速、简单和有效的策略。PCC 6803。
Cyanobacteria are simple, efficient, genetically-tractable photosynthetic microorganisms which in principle represent ideal biocatalysts for CO2 capture and conversion. However, in practice, genetic instability and low productivity are key, linked problems in engineered cyanobacteria. We took a massively parallel approach, generating and characterising libraries of synthetic promoters and RBSs for the cyanobacterium Synechocystis sp. PCC 6803, and assembling a sparse combinatorial library of millions of metabolic pathway-encoding construct variants. Genetic instability was observed for some variants, which is expected when variants cause metabolic burden. Surprisingly however, in a single combinatorial round without iterative optimisation, 80% of variants chosen at random and cultured photoautotrophically over many generations accumulated the target terpenoid lycopene from atmospheric CO2, apparently overcoming genetic instability. This large-scale parallel metabolic engineering of cyanobacteria provides a new platform for development of genetically stable cyanobacterial biocatalysts for sustainable light-driven production of valuable products directly from CO2, avoiding fossil carbon or competition with food production. Combinatorial assembly of libraries of metabolic pathway-encoding constructs from synthetic parts with suitable properties, followed by a small amount of screening, is a rapid, simple and effective strategy to obtain productive and genetically stable pathway designs for the cyanobacterium Synechocystis sp. PCC 6803.
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影响因子：
6.3
作者：
Du W;Jongbloets JA;van Boxtel C;Pineda Hernández H;Lips D;Oliver BG;Hellingwerf KJ;Branco Dos Santos F
通讯作者：
Branco Dos Santos F
影响因子：
4.4
作者：
Bakke, Ingrid;Berg, Laila;Valla, Svein
通讯作者：
Valla, Svein
影响因子：
3.7
作者：
Englund E;Pattanaik B;Ubhayasekera SJ;Stensjö K;Bergquist J;Lindberg P
通讯作者：
Lindberg P
影响因子：
4.7
作者：
Englund E;Andersen-Ranberg J;Miao R;Hamberger B;Lindberg P
通讯作者：
Lindberg P
影响因子：
5
作者：
Formighieri, Cinzia;Melis, Anastasios
通讯作者：
Melis, Anastasios