Adaptive Laboratory Evolution of Cupriavidus necator H16 for Carbon Co-Utilization with Glycerol
| Autor: | Hemanshi Galaiya, Tuck Seng Wong, Kang Lan Tee, Miriam Gonzalez-Villanueva, Paul Staniland, Ian Savill, Jessica Staniland |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Glycerol kinase Cupriavidus necator 030106 microbiology Heterotroph biodiesel glycerol engineering.material Catalysis Industrial waste lcsh:Chemistry Inorganic Chemistry Polyhydroxybutyrate 03 medical and health sciences chemistry.chemical_compound biopolymer Ralstonia eutropha H16 Glycerol Food science Physical and Theoretical Chemistry carbon co-utilization lcsh:QH301-705.5 Molecular Biology Cupriavidus necator H16 Spectroscopy Biodiesel adaptive evolution fat splitting biology Chemistry Organic Chemistry General Medicine biology.organism_classification Computer Science Applications 030104 developmental biology lcsh:Biology (General) lcsh:QD1-999 engineering lipids (amino acids peptides and proteins) Biopolymer |
| Zdroj: | International Journal of Molecular Sciences Volume 20 Issue 22 International Journal of Molecular Sciences, Vol 20, Iss 22, p 5737 (2019) |
| ISSN: | 1422-0067 1661-6596 |
| DOI: | 10.3390/ijms20225737 |
| Popis: | Cupriavidus necator H16 is a non-pathogenic Gram-negative betaproteobacterium that can utilize a broad range of renewable heterotrophic resources to produce chemicals ranging from polyhydroxybutyrate (biopolymer) to alcohols, alkanes, and alkenes. However, C. necator H16 utilizes carbon sources to different efficiency, for example its growth in glycerol is 11.4 times slower than a favorable substrate like gluconate. This work used adaptive laboratory evolution to enhance the glycerol assimilation in C. necator H16 and identified a variant (v6C6) that can co-utilize gluconate and glycerol. The v6C6 variant has a specific growth rate in glycerol 9.5 times faster than the wild-type strain and grows faster in mixed gluconate&ndash glycerol carbon sources compared to gluconate alone. It also accumulated more PHB when cultivated in glycerol medium compared to gluconate medium while the inverse is true for the wild-type strain. Through genome sequencing and expression studies, glycerol kinase was identified as the key enzyme for its improved glycerol utilization. The superior performance of v6C6 in assimilating pure glycerol was extended to crude glycerol (sweetwater) from an industrial fat splitting process. These results highlight the robustness of adaptive laboratory evolution for strain engineering and the versatility and potential of C. necator H16 for industrial waste glycerol valorization. |
| Databáze: | OpenAIRE |
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