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| Status |
Public on Jun 12, 2024 |
| Title |
Autotrophic adaptive laboratory evolution of the acetogen Clostridium autoethanogenum delivers the gas-fermenting strain LAbrini with superior growth, products, and robustness |
| Organism |
Clostridium autoethanogenum |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Microbes able to convert gaseous one-carbon (C1) waste feedstocks are increasingly important to transition to the sustainable production of renewable chemicals and fuels. Acetogens are interesting biocatalysts since gas fermentation using Clostridium autoethanogenum has been commercialised. However, most acetogen strains need complex nutrients, display slow growth, and are not robust for bioreactor fermentations. In this work, we used three different and independent adaptive laboratory evolution (ALE) strategies to evolve the wild-type C. autoethanogenum to grow faster, without yeast extract and to be robust in operating continuous bioreactor cultures. Multiple evolved strains with improved phenotypes were isolated on minimal media with one strain, named “LAbrini”, exhibiting superior performance regarding the maximum specific growth rate, product profile, and robustness in continuous cultures. Whole-genome sequencing of the evolved strains identified 25 mutations. Of particular interest are two genes that acquired seven different mutations across the three ALE strategies, potentially as a result of convergent evolution. Reverse genetic engineering of mutations in potentially sporulation-related genes CLAU_3129 (spo0A) and CLAU_1957 recovered all three superior features of our ALE strains through triggering significant proteomic rearrangements. This work provides a robust C. autoethanogenum strain “LAbrini” to accelerate phenotyping and genetic engineering and to better understand acetogen metabolism.
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| Overall design |
RNA sequencing of various strains of Clostridium autoethanogenum chemostat cultures grown on CO at various dilution rates; 15 samples in total with various number of biological replicate samples
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| Contributor(s) |
Ingelman H, Heffernan JK, Harris A, Brown SD, Shaikh KM, Yar Saqib A, Pinheiro MJ, de Lima LA, Rodriguez Martinez K, Gonzalez-Garcia RA, Hawkins G, Daleiden J, Tran L, Zeleznik H, Jensen RO, Reynoso V, Schindel H, Jänes J, Simpson SD, Köpke M, Marcellin E, Valgepea K |
| Citation(s) |
38871051 |
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| Submission date |
Feb 12, 2023 |
| Last update date |
Sep 12, 2024 |
| Contact name |
Kaspar Valgepea |
| E-mail(s) |
kaspar.valgepea@ut.ee
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| Phone |
+37256919831
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| Organization name |
University of Tartu
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| Department |
Institute of Technology
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| Lab |
ERA Chair in Gas Fermentation Technologies
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| Street address |
Nooruse 1
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| City |
Tartu |
| State/province |
Tartumaa |
| ZIP/Postal code |
50411 |
| Country |
Estonia |
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| Platforms (1) |
| GPL24458 |
Illumina NextSeq 500 (Clostridium autoethanogenum) |
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| Samples (15)
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| Relations |
| BioProject |
PRJNA934057 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE225123_Compiled_RPKM.txt.gz |
70.2 Kb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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