Fine-Tuned Protein Production in Methanosarcina acetivorans C2A,ACS Synthetic Biology

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Fine-Tuned Protein Production in Methanosarcina acetivorans C2A
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-06-19 00:00:00 , DOI: 10.1021/acssynbio.8b00062
Ann A. Karim ₁ , Daniel R. Gestaut ₂ , Maeva Fincker ₁ , John C. Ruth ₃ , Eric C. Holmes ₃ , Wayne Sheu ₃ , Alfred M. Spormann _{1,

3}

Affiliation

Methanogenic archaea can be integrated into a sustainable, carbon-neutral cycle for producing organic chemicals from C₁ compounds if the rate, yield, and titer of product synthesis can be improved using metabolic engineering. However, metabolic engineering techniques are limited in methanogens by insufficient methods for controlling cellular protein levels. We conducted a systematic approach to tune protein levels in Methanosarcina acetivorans C2A, a model methanogen, by regulating transcription and translation initiation. Rationally designed core promoter and ribosome binding site mutations in M. acetivorans C2A resulted in a predicable change in protein levels over a 60 fold range. The overall range of protein levels was increased an additional 3 fold by introducing the 5′ untranslated region of the mcrB transcript. This work demonstrates a wide range of precisely controlled protein levels in M. acetivorans C2A, which will help facilitate systematic metabolic engineering efforts in methanogens.

中文翻译：

乙酰甲烷单孢菌C2A的微调蛋白质生产

如果可以使用代谢工程技术改善产甲烷合成的速率，收率和效价，则产甲烷古菌可以整合到一个可持续的，碳中和的循环中，以从C ₁化合物生产有机化学品。但是，代谢工程技术由于无法控制细胞蛋白质水平的方法而在产甲烷菌中受到限制。我们进行了系统的方法，通过调节转录和翻译起始，来调节模式甲烷氧化甲烷菌（Methanosarcina acetivorans C2A）中的蛋白质水平。合理设计的食虫支原体中的核心启动子和核糖体结合位点突变C2A导致蛋白质水平的可预测变化超过60倍。通过引入mcrB转录本的5'非翻译区，蛋白质水平的总体范围又增加了3倍。这项工作证明了乙酸食甲烷支原体C2A中广泛的精确控制的蛋白质水平，这将有助于促进产甲烷菌中系统的代谢工程工作。

更新日期：2018-06-19

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