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However, the detailed metabolic mechanism by which sodium citrate promotes SAM production in\n                    <jats:italic>Pichia pastoris<\/jats:italic>\n                    has not yet been elucidated. Here, a comprehensive time\u2010series transcriptomic analysis revealed that sodium citrate profoundly influences gene expression across multiple pathways. Sodium citrate supplementation redirects carbon flux toward oxidative energy metabolism by upregulating amino acid biosynthesis, translation, and glycolysis. Crucially, the transcriptional upregulation of argininosuccinate synthase (\n                    <jats:italic>ARG1<\/jats:italic>\n                    ) and argininosuccinate lyase (\n                    <jats:italic>ARG4<\/jats:italic>\n                    ) was identified as a key node driving fumarate\u2010mediated TCA cycle anaplerosis and ATP supply. This foundational analysis led to the identification of\n                    <jats:italic>ARG4<\/jats:italic>\n                    as a key metabolic engineering target. Overexpression of\n                    <jats:italic>ARG4<\/jats:italic>\n                    significantly improved SAM production, achieving a 56.46% increase in shake flasks and 9.06\u00a0g\/L SAM (a 22.10% improvement) in 500\u00a0mL fermenters compared to control strains. Integrated physiological and metabolic flux analysis (MFA) demonstrated that\n                    <jats:italic>ARG4<\/jats:italic>\n                    overexpression redirects metabolic flow, channeling energy substrates toward SAM synthesis, effectively maintaining cellular respiratory metabolism and alleviating energy limitations in the late fermentation phase. This work establishes a novel strategy for enhancing product synthesis by restructuring energy allocation rather than merely increasing overall energy supply.\n                  <\/jats:p>","DOI":"10.1002\/biot.70304","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T12:19:17Z","timestamp":1788351557000},"source":"Crossref","is-referenced-by-count":0,"title":["Metabolic Engineering of\n                    <i>Pichia pastoris<\/i>\n                    for S\u2010Adenosylmethionine Overproduction via Transcriptomic\u2010Guided Energy Reallocation"],"prefix":"10.1002","volume":"21","author":[{"given":"Yuan\u2010Yuan","family":"Shang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Bioreactor Engineering East China University of Science and Technology  Shanghai People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhong\u2010Yue","family":"Li","sequence":"additional","affiliation":[{"name":"State 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