Multilevel Metabolic Engineering of Cupriavidus necator for CO₂-Derived Single-Cell Protein Production.

Authors

Menghan Chang, Zhiguo Huang, Shiqi Liu, Beini Yang, Ranran Wu, Lijuan Ma, Lei Chen, Zhiguang Zhu

Published in

Biotechnology journal. Volume 21. Issue 10. Pages e70320.

Abstract

Enhancing nitrogen assimilation and coordinating carbon-energy metabolism in the autotrophic chassis Cupriavidus necator remain key challenges for maximizing single-cell protein (SCP) production from CO₂. In this study, multilevel engineering was performed on C. necator. First, glnA was overexpressed to enhance nitrogen assimilation, which increased crude protein content to 72.75% in the best-performing strain, GS13. Preliminary physiological characterization suggested that this increase correlated with enhanced heme level. Second, glcR was disrupted to alter carbon allocation toward amino acid precursor metabolism, and epr-1 was deleted to inhibit extracellular protein degradation during the secretion process. The double-knockout strain Δepr-1ΔglcR achieved a high crude protein content of 70.13%. However, when the GS13 module was introduced into this background, OD₆₀₀ increased to 13.12 while crude protein content decreased to 68.05%, indicating a non-additive interaction between the engineered modules. Overall, our study provided improved strains and a modular framework for future SCP optimization and mechanistic studies of carbon-nitrogen coordination in C. necator.

PMID:
42875987
Bibliographic data and abstract were imported from PubMed on 15 Oct 2026.

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