ArticleMicrobial cell factories2026
Engineering serine metabolism to enhance AOX1 promoter self-induction in formate dehydrogenase-deficient Komagataella phaffii.
Article in Microbial cell factories, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.
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2 citing papers in PubMed.
- Metabolic engineering of Escherichia coli for methanol-assisted resveratrol biosynthesis.Bioprocess and biosystems engineering · 2026Article
- Evolutionary Adaptation and Targeted Metabolic Engineering Synergistically Improve Methanol-Derived Single-Cell Protein Production in Pichia pastoris.Biotechnology journal · 2026Article
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3 authors.
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Abstract
backgroundThe methylotrophic yeast Komagataella phaffii is a premier host for recombinant protein (rProt) production, which traditionally relies on methanol induction of the alcohol oxidase 1 promoter (PAOX1). However, the flammability and associated industrial limitations of methanol have motivated the search for methanol-free induction systems. We recently demonstrated that disruption of formate dehydrogenase gene (FDH) in K. phaffii allows endogenous formate derived from tetrahydrofolate (THF)-mediated C1 metabolism to induce PAOX1 without the addition of external inducers. Therefore, we hypothesized that increasing intracellular formate production by enhancing serine biosynthesis could further improve promoter induction and rProt productivity.
resultsOverexpression of SER3, encoding 3-phosphoglycerate dehydrogenase, the rate-limiting enzyme in serine synthesis, significantly increased PAOX1-driven expression of an intracellular reporter protein (eGFP) and secreted glucose oxidase (Gox) from Aspergillus niger without compromising cell fitness. Enhanced formate accumulation and stronger PAOX1 induction were observed in both microbioreactor and bioreactor cultivations using sorbitol or glycerol-sorbitol mixtures. In GOX-mSER3 strain grown in bioreactor, SER3 overexpression led to a 30% increase in specific Gox activity compared with that of the parental FdhKO strain.
conclusionsThis study provides a cost-effective metabolic engineering strategy for methanol-free, self-inducible expression systems in K. phaffii based on PAOX1, enabling safer and more sustainable industrial rProt production.
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