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TitleYeast Pathway Kit: A Method for Metabolic Pathway Assembly with Automatically Simulated Executable Documentation
Author(s)Pereira, Filipa Alexandra Barroso
Azevedo, Flávio Humberto Torres Dias Feio
Parachin, Nadia Skorupa
Hahn-Hagerdal, Barbel
Gorwa-Grauslund, Marie F.
Johansson, Björn
KeywordsMetabolic engineering
Saccharomyces cerevisiae
Synthetic biology
Issue date1-May-2016
CitationPereira, F., Azevedo, F., Parachin, N. S., Hahn-Hägerdal, B., Gorwa-Grauslund, M. F., & Johansson, B. (2016). Yeast pathway kit: a method for metabolic pathway assembly with automatically simulated executable documentation. ACS synthetic biology, 5(5), 386-394
Abstract(s)We have developed the Yeast Pathway Kit (YPK) for rational and random metabolic pathway assembly in Saccharomyces cerevisiae using reusable and redistributable genetic elements. Genetic elements are cloned in a suicide vector in a rapid process that omits PCR product purification. Single-gene expression cassettes are assembled in vivo using genetic elements that are both promoters and terminators (TP). Cassettes sharing genetic elements are assembled by recombination into multigene pathways. A wide selection of prefabricated TP elements makes assembly both rapid and inexpensive. An innovative software tool automatically produces detailed self-contained executable documentation in the form of pydna code in the narrative Jupyter notebook format to facilitate planning and sharing YPK projects. A D-xylose catabolic pathway was created using YPK with four or eight genes that resulted in one of the highest growth rates reported on D-xylose (0.18 h(-1)) for recombinant S. cerevisiae without adaptation. The two-step assembly of single-gene expression cassettes into multigene pathways may improve the yield of correct pathways at the cost of adding overall complexity, which is offset by the supplied software tool.
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Appears in Collections:DBio - Artigos/Papers

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