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Adaptive Laboratory Evolution of Cupriavidus necator H16 for Carbon Co-Utilization with Glycerol
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Document Title
Adaptive Laboratory Evolution of Cupriavidus necator H16 for Carbon Co-Utilization with Glycerol
Author
Gonzalez-Villanueva M, Galaiya H, Staniland P, Staniland J, Savill I, Wong TS, Tee KL
Name from Authors Collection
Affiliations
University of Sheffield; University of Sheffield; National Science & Technology Development Agency - Thailand; National Center Genetic Engineering & Biotechnology (BIOTEC)
Type
Article
Source Title
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Year
2019
Volume
20
Issue
5
Open Access
Green Accepted, Green Published, gold
Publisher
MDPI
DOI
10.3390/ijms20225737
Format
Abstract
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-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.
Funding Sponsor
Department of Chemical and Biological Engineering; ChELSI; EPSRC [EP/E036252/1]; BBSRC [BB/R506035/1]; RAEng\The Leverhulme Trust Senior Research Fellowship; BIOTEC Visiting Professorship; University of Sheffield Global Challenge Fellowship; CONACYT (Mexico) scholarship
License
CC BY
Rights
Authors
Publication Source
WOS