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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
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从可持续的碳来源最大化微生物生物生产,使用代系统工程.

Thomas Eng1, Deepanwita Banerjee1, Javier Menasalvas1

  • 1The Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Emeryville, CA 94608, USA; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.

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概括

我们设计了Pseudomonas putida用于生长合的印地戈因的生产,从para-coumarate获得高产. 这种系统生物学方法可以适应各种生物制造应用.

关键词:
艾利·艾利 (ALE) 是一个CP: 微生物学 微生物学这就是CRISPR/重组组合.伪omonas putida KT244040 的时间生物生产生物制造基因组规模的代谢模型增长合 增长合印地基氨酸 (indigoidine) 是一种红色的线性蛋白质 (lignin) 是一种蛋白质组学分析应变工程是一种应变工程.

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科学领域:

  • 合成生物学 合成生物学
  • 代谢工程是代谢工程.
  • 生物技术是生物技术.

背景情况:

  • 为了最大限度地提高异质生物分子的产生,需要了解细胞代谢和调节.
  • 增长合策略提高产品标题,产量和生产率.
  • 由于代谢模型的局限性,将这些策略应用于非正规碳来源受到阻碍.

研究的目的:

  • 开发一种增长合系统,用于在Pseudomonas putida KT2440中使用para-coumarate产生印地基丁.
  • 克服与非正规碳流和代谢模型差距相关的挑战.

主要方法:

  • 代设计-构建-测试-学习周期超过四次代.
  • 探索4,114种潜在的增长合解决方案.
  • 实验室进化和集体数据驱动的方法用于细菌菌株的精细化.
  • 功能性基因组学和实验验证.

主要成果:

  • 工程化Pseudomonas putida KT2440用于从para-coumarate中生长合的印地基因生产.
  • 实现了7.3g/L的印地基丁产量,最高理论产量为77%.
  • 证明了代设计和数据驱动改进的有效性.

结论:

  • 开发的增长合策略非常有效,并且可以在不同的宿主,碳流和产品中推广.
  • 这种系统层面的方法为优化生物制造工艺提供了一个强大的框架.
  • 这项研究强调了将计算设计与实验验证相结合的力量.