Phylogenetic debugging of a complete human biosynthetic pathway transplanted into yeast
| العنوان: | Phylogenetic debugging of a complete human biosynthetic pathway transplanted into yeast |
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| المؤلفون: | Jef D. Boeke, Paolo Mita, Neta Agmon, Itai Yanai, Maayan Baron, Tobias Schraink, Zuojian Tang, James A. Martin, Jun Chen, Jasmine Temple, David Fenyö, Benjamin P. Tu |
| المصدر: | Nucleic Acids Research |
| بيانات النشر: | Oxford University Press, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | Carboxy-Lyases, Saccharomyces cerevisiae, Biology, Chromosomes, Artificial, Human, chemistry.chemical_compound, Genetics, Humans, Amino Acid Sequence, Peptide Synthases, Gene, Phylogeny, Sequence Homology, Amino Acid, Adenine, Genetic Complementation Test, biology.organism_classification, Yeast, DNA shuffling, Biosynthetic Pathways, Transplantation, Isoenzymes, Metabolic pathway, chemistry, Biochemistry, Amidophosphoribosyltransferase, CRISPR-Cas Systems, Synthetic Biology and Bioengineering, Genetic Engineering, Sequence Alignment, DNA, Plasmids |
| الوصف: | Cross-species pathway transplantation enables insight into a biological process not possible through traditional approaches. We replaced the enzymes catalyzing the entire Saccharomyces cerevisiae adenine de novo biosynthesis pathway with the human pathway. While the ‘humanized’ yeast grew in the absence of adenine, it did so poorly. Dissection of the phenotype revealed that PPAT, the human ortholog of ADE4, showed only partial function whereas all other genes complemented fully. Suppressor analysis revealed other pathways that play a role in adenine de-novo pathway regulation. Phylogenetic analysis pointed to adaptations of enzyme regulation to endogenous metabolite level ‘setpoints’ in diverse organisms. Using DNA shuffling, we isolated specific amino acids combinations that stabilize the human protein in yeast. Thus, using adenine de novo biosynthesis as a proof of concept, we suggest that the engineering methods used in this study as well as the debugging strategies can be utilized to transplant metabolic pathway from any origin into yeast. |
| اللغة: | English |
| تدمد: | 1362-4962 0305-1048 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7e7f35911059c25100de9f065930e22a http://europepmc.org/articles/PMC7145547 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....7e7f35911059c25100de9f065930e22a |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13624962 03051048 |
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