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In this study, multilevel engineering was performed on\n                    <jats:italic>C. necator<\/jats:italic>\n                    . First,\n                    <jats:italic>glnA<\/jats:italic>\n                    was overexpressed to enhance nitrogen assimilation, which increased crude protein content to 72.75% in the best\u2010performing strain, GS13. Preliminary physiological characterization suggested that this increase correlated with enhanced heme level. Second,\n                    <jats:italic>glcR<\/jats:italic>\n                    was disrupted to alter carbon allocation toward amino acid precursor metabolism, and\n                    <jats:italic>epr\u20101<\/jats:italic>\n                    was deleted to inhibit extracellular protein degradation during the secretion process. The double\u2010knockout strain \u0394\n                    <jats:italic>epr\u20101<\/jats:italic>\n                    \u0394\n                    <jats:italic>glcR<\/jats:italic>\n                    achieved a high crude protein content of 70.13%. However, when the GS13 module was introduced into this background, OD\n                    <jats:sub>600<\/jats:sub>\n                    increased to 13.12 while crude protein content decreased to 68.05%, indicating a non\u2010additive interaction between the engineered modules. Overall, our study provided improved strains and a modular framework for future SCP optimization and mechanistic studies of carbon\u2010nitrogen coordination in\n                    <jats:italic>C. necator<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1002\/biot.70320","type":"journal-article","created":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T04:55:13Z","timestamp":1792040113000},"source":"Crossref","is-referenced-by-count":0,"title":["Multilevel Metabolic Engineering of\n                    <i>Cupriavidus necator<\/i>\n                    for CO\n                    <sub>2<\/sub>\n                    \u2010Derived Single\u2010Cell Protein Production"],"prefix":"10.1002","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-9962-0210","authenticated-orcid":false,"given":"Menghan","family":"Chang","sequence":"first","affiliation":[{"name":"College of Biotechnology Tianjin University of Science and Technology  Tianjin P. R. 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