Abstract：
:The Design-Build-Test-Learn (DBTL) cycle, facilitated by exponentially improving capabilities in synthetic biology, is an increasingly adopted metabolic engineering framework that represents a more systematic and efficient approach to strain development than historical efforts in biofuels and biobased products. Here, we report on implementation of two DBTL cycles to optimize 1-dodecanol production from glucose using 60 engineered Escherichia coli MG1655 strains. The first DBTL cycle employed a simple strategy to learn efficiently from a relatively small number of strains (36), wherein only the choice of ribosome-binding sites and an acyl-ACP/acyl-CoA reductase were modulated in a single pathway operon including genes encoding a thioesterase (UcFatB1), an acyl-ACP/acyl-CoA reductase (Maqu_2507, Maqu_2220, or Acr1), and an acyl-CoA synthetase (FadD). Measured variables included concentrations of dodecanol and all proteins in the engineered pathway. We used the data produced in the first DBTL cycle to train several machine-learning algorithms and to suggest protein profiles for the second DBTL cycle that would increase production. These strategies resulted in a 21% increase in dodecanol titer in Cycle 2 (up to 0.83 g/L, which is more than 6-fold greater than previously reported batch values for minimal medium). Beyond specific lessons learned about optimizing dodecanol titer in E. coli, this study had findings of broader relevance across synthetic biology applications, such as the importance of sequencing checks on plasmids in production strains as well as in cloning strains, and the critical need for more accurate protein expression predictive tools.
journal_name
ACS Synth Bioljournal_title
ACS synthetic biologyauthors
Opgenorth P,Costello Z,Okada T,Goyal G,Chen Y,Gin J,Benites V,de Raad M,Northen TR,Deng K,Deutsch S,Baidoo EEK,Petzold CJ,Hillson NJ,Garcia Martin H,Beller HRdoi
10.1021/acssynbio.9b00020subject
Has Abstractpub_date
2019-06-21 00:00:00pages
1337-1351issue
6issn
2161-5063journal_volume
8pub_type
杂志文章abstract：:Author: Recently, increasing research in siderophores has been dedicated to their possible medical applications in diagnostics and therapeutics for human pathogenic infections. Fusarinine C (FsC) is a natural hydroxamate siderophore that harbors three amino groups, which allow the easy chemical modification of FsC for...
journal_title：ACS synthetic biology
pub_type： 杂志文章
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abstract：:Genome engineering technologies now enable precise manipulation of organism genotype, but can be limited in scalability by their design requirements. Here we describe Merlin ( http://merlincad.org ), an open-source web-based tool to assist biologists in designing experiments using multiplex automated genome engineerin...
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journal_title：ACS synthetic biology
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abstract：:Cell-free systems, as part of the synthetic biology field, have become a critical platform in biological studies. However, there is a lack of research into developing a switch for a dynamical control of the transcriptional and translational process. The optogenetic tool has been widely proven as an ideal control switc...
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journal_title：ACS synthetic biology
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更新日期：2020-12-18 00:00:00
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journal_title：ACS synthetic biology
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更新日期：2018-10-19 00:00:00
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journal_title：ACS synthetic biology
pub_type： 信件
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更新日期：2018-03-16 00:00:00
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journal_title：ACS synthetic biology
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journal_title：ACS synthetic biology
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更新日期：2018-05-18 00:00:00
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pub_type： 信件
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journal_title：ACS synthetic biology
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