Development of a genetic toolset for the highly engineerable and metabolically versatile Acinetobacter baylyi ADP1
| Autor: | Biggs, Bradley W., Bedore, Stacy R., Arvay, Erika, Huang, Shu, Subramanian, Harshith, McIntyre, Emily A., Duscent-Maitland, Chantel V., Neidle, Ellen L., Tyo, Keith E.J. |
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| Rok vydání: | 2019 |
| Předmět: |
Acinetobacter baylyi ADP1
0303 health sciences Acinetobacter AcademicSubjects/SCI00010 Cas9 030306 microbiology Genomics Computational biology Biology Lignin Genome Protein expression Metabolic engineering 03 medical and health sciences Synthetic biology Metabolic Engineering Genetics Cloning Molecular Bioprocess Promoter Regions Genetic Synthetic Biology and Bioengineering Ribosomes Genome Bacterial 030304 developmental biology |
| Zdroj: | Nucleic Acids Research |
| DOI: | 10.1101/696302 |
| Popis: | One primary objective of synthetic biology is to improve the sustainability of chemical manufacturing. Naturally occurring biological systems can utilize a variety of carbon sources, including waste streams that pose challenges to traditional chemical processing, such as lignin biomass, providing opportunity for remediation and valorization of these materials. Success, however, depends on identifying micro-organisms that are both metabolically versatile and engineerable. Identifying organisms with this combination of traits has been a historic hindrance. Here, we leverage the facile genetics of the metabolically versatile bacterium Acinetobacter baylyi ADP1 to create easy and rapid molecular cloning workflows, including a Cas9-based single-step marker-less and scar-less genomic integration method. In addition, we create a promoter library, ribosomal binding site (RBS) variants and test an unprecedented number of rationally integrated bacterial chromosomal protein expression sites and variants. At last, we demonstrate the utility of these tools by examining ADP1’s catabolic repression regulation, creating a strain with improved potential for lignin bioprocessing. Taken together, this work highlights ADP1 as an ideal host for a variety of sustainability and synthetic biology applications. |
| Databáze: | OpenAIRE |
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