التفاصيل البيبلوغرافية
| العنوان: |
Exometabolomics Assisted Design and Validation of Synthetic Obligate Mutualism |
| المؤلفون: |
Adam M. Deutschbauer, Suzanne M. Kosina, Jayashree Ray, Adam P. Arkin, Yumi Suh, Megan A. Danielewicz, Anup K. Singh, Suzan Yilmaz, Mujahid Mohammed, Trent R. Northen |
| المصدر: |
Kosina, SM; Danielewicz, MA; Mohammed, M; Ray, J; Suh, Y; Yilmaz, S; et al.(2016). Exometabolomics Assisted Design and Validation of Synthetic Obligate Mutualism. ACS Synthetic Biology, 5(7), 569-576. doi: 10.1021/acssynbio.5b00236. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/2019585b |
| بيانات النشر: |
eScholarship, University of California, 2016. |
| سنة النشر: |
2016 |
| مصطلحات موضوعية: |
0301 basic medicine, Transposable element, Auxotrophy, 030106 microbiology, Mutant, Microbial Consortia, Biomedical Engineering, Computational biology, medicine.disease_cause, Biochemistry, Genetics and Molecular Biology (miscellaneous), Zymomonas mobilis, 03 medical and health sciences, Microbial ecology, medicine, Escherichia coli, Metabolomics, Amino Acids, Symbiosis, Mutualism (biology), Zymomonas, biology, Obligate, business.industry, Escherichia coli Proteins, Reproducibility of Results, General Medicine, biology.organism_classification, Coculture Techniques, Biotechnology, 030104 developmental biology, Mutation, Synthetic Biology, business |
| الوصف: |
© 2016 American Chemical Society. Synthetic microbial ecology has the potential to enhance the productivity and resiliency of biotechnology processes compared to approaches using single isolates. Engineering microbial consortia is challenging; however, one approach that has attracted significant attention is the creation of synthetic obligate mutualism using auxotrophic mutants that depend on each other for exchange or cross-feeding of metabolites. Here, we describe the integration of mutant library fitness profiling with mass spectrometry based exometabolomics as a method for constructing synthetic mutualism based on cross-feeding. Two industrially important species lacking known ecological interactions, Zymomonas mobilis and Escherichia coli, were selected as the test species. Amino acid exometabolites identified in the spent medium of Z. mobilis were used to select three corresponding E. coli auxotrophs (proA, pheA and IlvA), as potential E. coli counterparts for the coculture. A pooled mutant fitness assay with a Z. mobilis transposon mutant library was used to identify mutants with improved growth in the presence of E. coli. An auxotroph mutant in a gene (ZMO0748) with sequence similarity to cysteine synthase A (cysK), was selected as the Z. mobilis counterpart for the coculture. Exometabolomic analysis of spent E. coli medium identified glutathione related metabolites as potentially available for rescue of the Z. mobilis cysteine synthase mutant. Three sets of cocultures between the Z. mobilis auxotroph and each of the three E. coli auxotrophs were monitored by optical density for growth and analyzed by flow cytometry to confirm high cell counts for each species. Taken together, our methods provide a technological framework for creating synthetic mutualisms combining existing screening based methods and exometabolomics for both the selection of obligate mutualism partners and elucidation of metabolites involved in auxotroph rescue. |
| وصف الملف: |
application/pdf |
| اللغة: |
English |
| URL الوصول: |
https://explore.openaire.eu/search/publication?articleId=doi_dedup___::47258f210d3b9d86a4d7fdb78db35943
http://www.escholarship.org/uc/item/2019585b |
| حقوق: |
OPEN |
| رقم الأكسشن: |
edsair.doi.dedup.....47258f210d3b9d86a4d7fdb78db35943 |
| قاعدة البيانات: |
OpenAIRE |
