Metabolic impact of redox perturbations on recombinant protein production in Komagataella phaffii under methanolic growth conditions

Metabolic impact of redox perturbations on recombinant protein production in Komagataella phaffii under methanolic growth conditions | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Metabolic impact of redox perturbations on recombinant protein production in Komagataella phaffii under methanolic growth conditions Eric Antón, Sergi Monforte, Ane Quesada-Ganuza, Joan Albiol, Pau Ferrer This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9453242/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 7 You are reading this latest preprint version Abstract Background Recombinant protein extracellular production is a highly demanding process in terms of energy and redox cofactor regeneration. Yeasts such as Komagataella phaffii (aka. Pichia pastoris ) seem to adapt their metabolism by increasing ATP, NADH or NADPH regeneration to compensate for the metabolic burden exerted by such costly process. Earlier studies have proven that increased NADPH regeneration capacity has a positive effect on recombinant protein production when using glucose as sole carbon source, while counteracting increased NADH/NAD + ratios has also been proven beneficial. Results To evaluate the metabolic impact of increasing NADPH and/or NAD + regeneration caused by the introduction of a cytosolic NADH kinase and an alternative NADH oxidase, we initially performed in silico simulations based on the use of a genome-scale model. Next, we investigated the effect of overexpressing the S. cerevisiae ’s POS5 NADH kinase encoding gene (targeted to the cytosol) and an alternative oxidase ( AtOX ) in a K. phaffii strain producing extracellularly a Rhizopus oryzae lipase (Rol) under control of the alcohol oxidase 1 ( AOX1 ) promoter (P AOX1 ), thereby assessing the impact of such engineering strategies on recombinant protein production and cell performance under methanol growth conditions. Small-scale cultures allowed us to conclude that such genetic modifications increase protein production significantly under methanol growth conditions. The redox-engineered strains were further characterized in bioreactor chemostat cultures using a glucose/methanol mix (60/40%, w/w) as a carbon source to corroborate their performance. Thereby, up to a 36% increase in Rol specific productivity was observed when overexpressing two copies of POS5 , whereas AtOX overexpression did not improve production. Notably, combined two-copy POS5 and AtOX overexpression resulted in a 53% increase in lipase production compared to the reference strain, suggesting that perturbation of the NADH/NAD + ratio leads to widespread changes in metabolism ultimately affecting cell growth and product formation, resulting in the observed additive effect. An in-silico interpretation of the physiological macroscopic data from chemostat cultures using a genome-scale metabolic model allowed us to gain further insights on the metabolic impact of these modifications. Conclusions Our study provides a DBTL cycle workflow generating valuable insights for a quantitative understanding and prediction of cellular response to redox- based perturbations and subsequent test bedding of metabolic engineering applications in K. phaffii . Komagataella phaffii NADH kinase alternative oxidase Rhizopus oryzae lipase POS5 AOX redox cofactor balance metabolic burden genome-scale metabolic model Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Overproduction of recombinant proteins in yeast such as Komagataella phaffii (also known as Pichia pastoris ), exerts a significant metabolic burden to the cell due to the drain of resources from cell maintenance and growth (1,2). This contributes to the metabolic stress caused by overexpression of recombinant proteins, negatively impacting productivity (2,3). Such metabolic burden may be triggered by several factors: de novo synthesis of energetically costly amino acids, overloaded protein folding machinery, posttranslational protein modifications and secretion processes. These lead to energetic and redox balance alterations and a suboptimal cell physiological state to support recombinant protein production. Specifically, production of recombinant proteins in K. phaffii leads to a redistribution of metabolic fluxes to increase supply of ATP and NADH (4–7). In addition, several 13 C-MFA studies in recombinant K. phaffii point at increased NADPH supply through pentose phosphate pathway (PPP) in production strains (8,9). Several metabolic engineering strategies targeting redox metabolism have been explored to overcome these limitations in K. phaffii (3). For instance, overexpression of genes encoding PPP enzymes increased NADPH supply, allowing for improved recombinant protein yield under glucose-growing conditions, using the constitutive GAP promoter (P GAP ) expression system (10). Similarly, we have demonstrated the positive effect of increasing NADPH availability by overexpressing a heterologous NADH kinase encoding POS5 gene from the yeast Saccharomyces cerevisiae (11) in a K. phaffii strain producing an antibody fragment under the control of P GAP , thereby increasing secreted product titers by 1.55 and 2-fold in normoxic and hypoxic glucose-limited chemostats, respectively. Notably, the use of glycerol as an alternative C-source resulted in a lower effect (12) . Overall, these studies strongly support the hypothesis of a higher limitation of recombinant protein production by NADPH when cells are grown on glucose, as well as pointing out that different redox, NADPH, engineering solutions may be needed depending on the growth conditions, as expected from the observation that sources of NADPH may vary depending on the C-source (13). This may be particularly relevant when the use of alternative C-sources such as methanol (or combinations thereof) is foreseen. Besides, the impact of NADH availability on K. phaffii growth and protein production has also been explored by overexpressing a heterologous NADH oxidase from Lactococcus lactis (14). By oxidizing NADH to NAD + , the NADH/NAD + ratio increase caused by recombinant protein production was counteracted. This resulted in an upregulation of methanol metabolism genes and recombinant CALB lipase gene (expressed under the control of the methanol-inducible AOX1 promoter), which in turn allowed to improve methanol uptake rate and recombinant protein production. Moreover, by further co-overexpressing an adenylate kinase ( ADK1 ) from S. cerevisiae , the ATP supply in this strain, which was reduced by the lower NADH availability for ATP synthesis, was restored, achieving even higher production levels. In the light of these studies and in order to develop a lean workflow to systematically assess the impact of different redox engineering strategies on NADPH regeneration and heterologous protein production in K. phaffii under methanolic growth conditions, a model-based Design-Build-Test-Learn (DBTL) cycle approach, enabled by 13 C-fluxomics, has been conceived and benchmarked with two different genetic targets and an exemplary well-defined growth condition. To this end, in this study, we have further investigated the effect of cofactor regeneration on the heterologous production of a Rhizopus oryzae lipase (Rol) and the central carbon metabolism of K. phaffii growing under methanolic conditions, by overexpressing a heterologous NADH kinase encoding gene POS5 from Saccharomyces cerevisiae and an alternative oxidase ( AtOX ) from Histoplasma capsulatum . The NADH kinase was targeted to the cytosol, providing an extra NADPH source, whereas the alternative oxidase is an enzyme located in the inner mitochondrial membrane that provides an alternative route to transfer electrons from the ubiquinol pool to oxygen, bypassing several proton-pumping steps this way. Similarly, this alternative oxidase has been shown to locate in mitochondria when ectopically overexpressed in S. cerevisiae (15). A previous study showed an important physiological impact when the endogenous alternative oxidase gene in K. phaffii was either overexpressed or eliminated (16), but its effect in recombinant protein production was not investigated. Firstly, genome scale model-based simulations were performed to assess the impact of increased flux through the ectopic reactions on both macroscopic growth parameters, NADPH regeneration and product formation under a defined set of growth conditions (mixed multicarbon source:methanol chemostat cultivations), for which previous 13 C-Metabolic Flux analyses ( 13 C-MFA) were available (7,17,18). Second, the redox-engineered strains were constructed and were tested at shake flask scale and then further characterized in 1-L scale chemostat cultures. Finally, to gain a deeper insight into the physiological effect of these modifications, an in silico interpretation of the metabolic effect of these genetic modifications was also performed. Materials and methods Strains and plasmids A K. phaffii X-33-derived strain containing a single copy of the gene encoding the lipase of Rhizopus oryzae ( ROL ) was used in this study (19). The expression cassette of this gene, constructed using the pPICZαA vector (Invitrogen-Thermo Fisher Scientific, California, USA), is under control of the methanol-inducible AOX1 promoter (P AOX1 ) and includes the encoding region of the S. cerevisiae α-mating factor secretion signal (20). The genes encoding the NADH kinase Pos5p ( POS5 ) from the yeast Saccharomyces cerevisiae and the alternative oxidase ( AtOX ) from the fungi Histoplasma capsulatum were codon-optimized for K. phaffii and synthetized by Geneart (ThermoFischer Scientific) and cloned into a pPUZZLE vector (21) under control of GAP promoter. In addition, the first 49 bp of POS5 ORF, encoding region of the mitochondrial signal peptide, were removed, i.e. ensuring both cytosolic and mitochondrial location of the corresponding protein product, and cloned in the pPUZZLE vector. Construction of POS5 and truncated POS5 expression vectors have been previously described in detail (12). Briefly, 2 vectors were constructed in this study: pPUZZLE_cPOS5 and pPUZZLE_ Hc AOX1 ( Supplementary figure I ). The single mutants were generated by genomic integration of the c POS5, mPOS5 and Hc AOX1 expression vectors . pPUZZLE plasmids were linearized using AvrII (New England Biolabs, Massachusetts, USA) and introduced into K. phaffii X-33/ROL using 100 ng of DNA. Transformation was performed by electroporation with a Gene PulserXcell™ Electroporation System (Bio-Rad, Hercules, CA, USA) using as parameters: 1550 V, 25 µF and 200 Ω. Positives clones were selected by growing on Yeast Peptone Dextrose (YPD) plates with 500 µg/mL of G418 (Geneticin) and checked by colony PCR (all primers used in this study are described in Supplementary table I ). The combined POS5 and Hc AtOX1 mutants (POSNOX) were obtained by transforming the strain harboring the alternative oxidase with 500 ng of linearized with AvrII BB3eH_14_pGAP_cPOS5_ScCYCtt plasmid (22). 200 ng/mL of Hygromycin was added on YPD plates as selection marker. Given that clonal variation was expected in the number of cPOS5 cassette copies integrated in the genome, around 20 positive clones were isolated and characterized. Electrocompetent K. phaffii X-33 cells were prepared according to established protocols (23). Strains constructed in this study are summarized in Table 1 . Table 1 List of strains constructed and used in this study Strain Parental strain Plasmids and cassettes used for transformation Genotype Antibiotic resistance Reference X-33 ROL X-33 pPICZa_ROL plasmid linearized with PmeI Rol-producing strain Zeocin 30 X-33 ROL cPOS5 X-33/ ROL pPUZZLE_cPOS5 linearized with AvrII Rol-producing strain harbouring 1 or more c POS5 (cytosolic POS5 ) copies G418 (Geneticin) and Zeocin 30 X-33 ROL HcAtOX X-33 ROL pPUZZLE_cPOS5 Linearized with AvrII Rol producing strain harbouring 1 or more cytosolic Hc AOX1 copies G418 (Geneticin) and Zeocin 30 X-33 ROL POSNOX X-33 HcAtOX BB3eH_14_pGAP_cPOS5_ScCYCtt Linearized with AvrII Rol-producing strain harbouring 1 or more c POS5 copies and 1 HcAOX1 copy G418 (Geneticin), Zeocin and Hygromycin This work Screening conditions To consider clonal variation of K. phaffii transformants, five colonies from each strain were selected and further tested in shake flask cultures in triplicate at 25ºC and 150 rpm, as described by Cámara and coworkers (19). Briefly, strains were first grown in 50 mL of Buffered Minimal Glycerol (BMG) medium in 1-L shake flasks overnight at 25ºC and 150 rpm. Afterwards, reinoculation from BMG into 25 mL of fresh Buffered Minimal Methanol (BMM) medium in 250-mL shake flasks was performed at an initial optical density of 1 and further grown under the same conditions. One pulse of 125 µL of pure methanol was added (0.5% v/v) after 24 h and 48 h of cultivation. Samples for biomass and recombinant protein quantification were taken after 70 h of cultivation. Chemostat cultivation conditions Selected strains were characterized in duplicate chemostat cultures using a mixed carbon source consisting of 60%:40% glucose:methanol (w/w). Cultivations were performed at a 700-mL working volume in a DASGIP Parallel Bioreactor System (Eppendorf, Germany). Operating conditions were set at 25°C, pH 5.0 controlled through the addition of 15% ammonia (v/v), 700 rpm, 1 vvm inlet gas flow, and a dilution rate (D) of 0.1 h⁻¹. Steady states were monitored along the 4th, 5th, and 6th residence times to ensure stable biomass concentration, CER, OUR, lipase activity and extracellular metabolite profiles. Measurements at the end of the 5th residence time were considered as experimental steady-state growth parameters. Batch medium contained: 19.95 g/L glycerol, 0.9 g/L citric acid, 6.3 g/L (NH 4 ) 2 HPO 4 , 0.01 g/L CaCl 2 , 0.45 g/L KCl, 0.25 g/L MgSO 4 ·7H 2 O, 1 mL/L Biotin (0.2 g/L; Sigma, Munich, Germany), 0.3 mL antifoam (Glanapon, Bussetti & Co GmbH, Wien, Austria) and 2.3 mL/L PTM1 trace salts stock solution. Chemostat medium contained: 25g/L total carbon source, 0.42 g/L citric acid, 2.18 g/L (NH 4 ) 2 HPO 4 , 5.5 mg/L CaCl 2 ·2H 2 O, 0.85 g/L KCl, 0.32 g/L MgSO 4 ·7H 2 O, 0.5 mL Biotin (0.2 g/L), 50 µL antifoam (Glanapon) and 0.8 mL PTM1 trace salts stock solution. Trace salt composition was: 6.0 g/L CuSO 4 .5H 2 O; 0.08 g/LNaI; 3.36 g/L MnSO 4 .H 2 O; 0.2 g/LNa 2 MoO 4 .2H 2 O; 0.02 g/L H 3 BO 3 ; 0.82 g/L CoCl 2 .6H 2 O; 20 g/L ZnCl 2 ; 65 g/L FeSO 4 .7H 2 O and 5.0 mL H 2 SO 4 (95–98% w/w). Medium pH was adjusted to 5 with 25% HCl. All chemostat cultivations were performed at least in duplicate. Analytical methods Biomass determination Biomass optical density was measured at 600 nm using a DR3900 spectrophotometer (Hach, Bizkaia, Spain). Dry cell weight (DCW) was determined using pre-weighed dried glass fiber filters (Millipore, Massachusetts, USA) as reported by Jordà and coworkers (24). Both measurements were performed in triplicate. Lipolytic activity assay Lipase activity in the cleared cultivation broth was quantified using a colorimetric assay based on the hydrolysis of p-nitrophenyl butyrate as substrate, following the method described by Guillén et al. (2011)(25). Metabolite quantification Residual glucose and methanol concentrations in the cleared cultivation broth were measured by means of HPLC. Specifically, a Vanquish Flex UHPLC system (Thermo Fisher Scientific, Waltham, MA, USA) equipped with an ICSep ICE-COREGEL 87H3 (Transgenomic) ion exchange column and a Water 2410 (Waters) refraction index detector. CROMELEON software (Dionex) was used for data analysis. Heterologous POS5 gene copy number determination To quantify the heterologous (from S. cerevisiae ) POS5 gene dosage, ddPCR analysis was performed. First, genomic DNA from POSNOX clones was extracted using a commercial kit (Wizard Genomic DNA Purification Kit, Promega, Madison, WI) following the manufacturer’s instructions and quantified in a NanoDrop 2000 spectrophotometer (Thermo Scientific). QX200 ddPCR EvaGreen Supermix reaction cocktail (Bio-Rad) was used to carry out the assay. Operational parameters can be consulted in the manufacter’s guideline for this product. Once reaction mixtures were prepared a QX200 Droplet Generator (Bio-Rad) was employed to generate droplets and the samples were transferred into a 96-well plate. After thermal cycling procedure, the plate was placed in a QX200 Droplet Reader (Bio-Rad). Results data acquisition was conducted with the software QuantaSoft v.1.7. The POS5 gene copy number was determined by calculating the ratio of positive droplets between the sample and a reference sample carrying one copy of POS5 . Prior temperature and DNA concentration gradient ddPCRs were performed to determine the optimal conditions for the reaction and the one POS5 Copy reference clone. For these tests a one copy housekeeping gene, namely β-Actin (ACT1) was selected to normalize data. The annealing temperature and the DNA quantity per sample were set to 58°C and 1 ng (or 0.05 ng/µL) respectively. Primers used for this assay are described in Supplementary table I . In silico metabolic flux analysis The iMT1026 v.3.0 genome scale model of K. phaffii (26) was used in the COBRApy 0.26.2 (27) toolbox with CPLEX 12.9 as solver. The simulations of the reference Rol-producing strain were done using pFBA, maximizing the biomass reaction and defining a set of reaction ratios extracted from previous 13 C-tracer experiments performed on mixed methanol and glucose or glycerol chemostat cultivations (7,18) as constraints depending on the simulated cultivation conditions (60/40% Glucose/Methanol or 60/40% Glycerol/Methanol, respectively). Simulations of the redox-engineered strains were performed using Non-Linear Minimization of Metabolic Adjustment (MOMA), with the wild-type simulations as reference conditions. For all simulations performed, the carbon source uptake rate was fixed to match the reference X-33 ROL strain cultivations experimental data. In the POS5 - and HcAtOX-overexpressing strains simulations, the ectopic reactions were added at fixed flux values ranging from 0 to 0.2 mmol/(gDCW·h) to explore its impact in the metabolic network. Redox cofactors regeneration was calculated by summing the absolute flux value of all the reaction involved in the consumption or generation of a cofactor and diving it by 2 as detailed elsewhere (28). Constrained based models do not include metabolite concentrations as state variables. Thus, redox cofactors ratios cannot be calculated. Therefore, we used metabolite turnover rates (AKA flux-sum (29) as a proxy for changes in redox balance. Metabolites flux-sum were calculated as described in Eq. 1: Eq. 1 Where S ij is the stoichiometric coefficient of metabolite i in reaction j and v j is the flux of reaction j. Statistical analyses Unless otherwise stated, data are expressed as mean ± standard deviation of at least triplicate samples. To test for significance when comparing experimental data, obtained from continuous cultures and transcriptomic experiments, a Student’s t -test was performed using the Microsoft Excel software. A p-value lower than 0.05 was considered statistically significant. Results and discussion In silico prediction of increased Pos5 and Aox1 activity impact on physiological parameters and recombinant protein production under different cultivation conditions (Design phase) We explored beforehand the potential effects of ectopically overexpressing the S. cerevisiae ’s c POS5 (truncated POS5 for cytosolic localization) or H. capsulatum ’s AOX1 (HcAtOX1) genes on the physiological growth parameters and intracellular fluxes distribution by performing genome-scale model-based simulations in which we increased iteratively the flux value of their corresponding reactions and fixing them as constraints. The resulting metabolic flux distribution was obtained by performing an NL MOMA, using as reference simulation a previous pFBA constrained with reaction ratios obtained from earlier 13 C-MFA studies (7,17,18). We conducted this procedure for both ectopic reactions separately under two different co-substrate feeding regimes under metabolic steady state conditions, that is, glucose or glycerol plus methanol in a fixed ratio (60% co-substrate + 40 % methanl, (w/w)). The simulations showed modest perturbations in the respiratory parameters (specific oxygen consumption and CO 2 production rates, q O2 and q CO2 ) when flux through the ectopic reactions was increased. Notably, while growth progressively diminished in all cases, production of the recombinant protein Rol increased notably in the POS5 case study (Fig. 1 ). In contrast, the introduction of HcAOX1 resulted in a negative effect on protein production as well as inducing minor respiratory adjustments. Overall, the simulation results depicted in Fig. 1 suggested that recombinant protein production is primarily impacted by cPOS5 overexpression, while HcAOX1 does not show a significant benefit, with respiration and final biomass remaining essentially unaffected, irrespective of the co-substrate (glucose or glycerol) used. In silico prediction of the impact of Pos5 and AtOX activity on redox cofactors turnover rates (Design phase) Given that both ectopic reactions are directly related to the redox pairs NADPH/NADP + and NADH/NAD + , the NAD(P)H regeneration capacity in the potential engineered strains was evaluated by calculating the turnover rates of these metabolites in the cytosol for every simulation previously performed. In the initial exploration (i.e., scanning ectopic flux from 0 to 0.2 mmol/(gDCW·h)), we observed an increasing tendency in regenerative capacity for all cofactors (Fig. 2 ), the NADPH/NADP + pair showing the most pronounced increases. Remarkably, this pattern was more prominent in the POS5 case, especially when glucose was used as co-substrate, coherent with the fact that Pos5 kinase reaction directly generates NADPH. However, considering that alternative oxidases boost fluxes through the electron transport chain subunits reactions, we expected that the impact on the NAD + cofactor regeneration rate would be higher when simulating the AtOX gene overexpression. This outcome could be attributed to the influence that the endogenous mitochondrial Pos5 kinase could have on the energetic state, as it consumes ATP to phosphorylate NADH. Coherently, simulations revealed increasing ATP demand under this scenario (Supplementary Fig. 2). These turnover rate calculations were done for the cytosolic pools of the cofactors, since metabolite pools are differentiated by subcellular location in the genome-scale model. Thereby, we also calculated the turnover rates for the redox cofactors located in the mitochondria (Supplementary Fig. 3), obtaining coherent values. Namely, a higher regeneration rate of mitochondrial NAD + and NADH when HcAOX is overexpressed in contrast to ectopic c POS5 expression. Moreover, the 2cPOS5 and HcAOX1 (POSNOX) combined stran was tested in silico by fixing the HcAOX1 reaction flux to the estimated value (0.1 mmol/(gDCW·h)) and progressively augmenting the POS5 reaction flux. This analysis revealed an additive effect in the NADPH regeneration capacity (Supplementary Fig. 4). Overall, the initial in silico exploration revealed that the expression of both ectopic genes would prove beneficial in the redox cofactors’ regeneration capacity of the engineered strains, thereby motivating subsequent strain construction and testing. Moreover, once the engineered strains characterization in chemostat cultures was completed, simulations were run again using the obtained experimental data as constraints to get new insights on how the metabolic flux could be redistributed because of POS5 and HcAtOX expression. In addition, the Glucose 60% / Methanol 40% (w/w) mixture was selected for further characterization of the engineered strains at bioreactor scale based on the following considerations: First, in silico predictions indicated that recombinant protein production more enhanced when glucose is used as a co‑substrate, compared to the glycerol case. This observation was coherent with previous chemostat cultivation studies using glycerol as co-substrate with the first series of redox engineered Rol-producing strains, where the observed increases in lipase activity relative to the reference strain were not statistically significant (30). Second, perturbations in redox cofactor balance caused by POS5 overexpression, i.e., on the NADPH/NADP + ratio, have been shown to be more pronounced when cells are cultivated on glucose compared to glycerol (31). Although using methanol as the sole carbon source was initially considered, literature reports indicate that similar induction levels can be achieved with mixed carbon sources (32). Additionally, methanol appears to function more effectively as an auxiliary energy substrate in K. phaffii (33). POS5 and Hc AtOX co-expression strain construction (Build) Electrocompetent strains carrying a single HcAtOX copy were transformed with a linearized pPuzzle vector containing the cPOS5 kinase. To quantify the number of integrated cPOS5 copies in the resulting transformants, ddPCR analysis was performed on several clones. Most isolates incorporated one, two, or three copies of the NADPH kinase encoding gene (Supplementary table 2), indicating efficient but variable integration of the construct. Small scale screening of POS5 and HcAtOX1 co-expressing clones (Test) All transformant clones were first cultivated at small scale in BMM medium, followed by assessment of lipase activity to estimate recombinant protein production. Under these screening conditions, all clones transformed with the c POS5 gene displayed no significant increment in recombinant protein production except for one individual clone harboring 2 copies of the ectopic gene. Interestingly, the majority of HcAtOX clones carrying more than two copies of the cytosolic POS5 kinase displayed a clear increase in lipase activity (Fig. 3 ). To further evaluate this set of engineered strains, only the clones harboring exactly two POS5 copies were selected for reactor‑scale characterization, as strains with higher copy numbers exhibited a pronounced reduction in growth rate during the inoculum preparation. Reactor-scale characterization of POS5 gene dosage effect and co-overexpression of POS5 and AtOX on recombinant protein production and physiological parameters. (Test) A strain co‑expressing the ectopic genes investigated in this study was constructed and evaluated in chemostat cultures using 60% (w/w) glucose as a co‑substrate to assess the predicted synergistic effect on protein production. Additionally, a strain harboring a single POS5 kinase copy was examined to verify that one copy of this gene does not enhance Rol synthesis, as observed from small‑scale experiments. Consequently, a new set of cultivations were performed using the reference X-33 ROL strain, HcAtOX, 1cPOS5, 2cPOS5, as well as a strain combining two POS5 copies with one HcAtOX copy. The 1cPOS5 strain did not enhance Rol production (Table 1 ), confirming that a single POS5 copy is insufficient to improve recombinant protein synthesis. In contrast, the HcAtOX strain exhibited a pronounced physiological impact, including a clear reduction in biomass yield and a hampering of methanol assimilation, as indicated by its lower methanol-specific consumption rate and elevated residual methanol concentrations. This strain also showed a decrease in Rol activity, demonstrating that the introduction of a single HcAtOX copy negatively affects both growth and protein production. Although neither 1cPOS5 nor 2cPOS5 strains substantially altered biomass yield or residual methanol levels, both POS5-based modifications increased respiratory activity, reflected by higher CO₂ secretion rates and elevated RQ values. Strains carrying two POS5 copies (2cPOS5), including the combined 2cPOS5–HcAtOX construct displayed substantial improvements in Rol production, reaching increases of approximately 30–40%. Notably, the additional HcAtOX copy in the 2cPOS5-HcAtOX combined mutant did not further enhance Rol synthesis relative to 2cPOS5, but it strongly influenced macroscopic process parameters, including reduced final biomass concentrations and pronounced increases in both oxygen uptake and CO₂ secretion rates. Table 1. Macroscopic parameters of the redox engineered and reference Rol-producing strains cultivated in chemostat using a glucose 60% methanol 40% mix as carbon source. Experimental data constrained in silico analysis of ectopic genes overexpression impact on internal metabolic fluxes (Learn phase) To further understand how the expression of the ectopic genes tweaks K. phaffii’s metabolism, bolstering the regeneration of redox cofactors and, ultimately, enhancing the recombinant protein production, simulations were repeated, setting the carbon source uptake rates obtained in chemostat cultivations as model constraints. The prediction results were visualized in a simplified K. phaffii metabolic network and further complemented with transcriptomics data of reporter genes previously obtained from analogous cultivations (30). Primarily, there is a redirection of flux to the methanol dissimilatory pathway which takes place regardless of the ectopic gene expressed (Fig. 4 ). This metabolic shift in the peroxisome is coherent with the increasing NADH regeneration rates seen in the initial in silico exploration and could also explain why the engineered strains’ growth rate is diminished. Another common feature in all simulations is the increase of upper and lower glycolysis reactions flux values leading to a greater pool of phosphoenolpyruvate. Such raise could be explained by the higher demand of energy spent on the ectopic kinase’s functioning or a general metabolic pull directed to the TCA cycle, as the oxidase may be enhancing the activity of the electron transport chain in the mitochondria. At the same time, differences in the internal fluxes’ perturbations caused by the expression of ectopic genes can be found too. For example, the oxidative branch reactions in the pentose phosphate pathway (PPP), one of the main NADPH sources in yeast (34), are upregulated in the AtOX simulations but their flux decreases when POS5 activates as the kinase is already providing NADPH to the network. This observation is further reinforced by transcriptomic data, which shows corresponding changes in gene expression and suggests a clear correlation between catalytic activity and transcriptional regulation. Lastly, the TCA cycle fluxes are perturbed differently. Particularly, the NAD-dependent Isocitrate dehydrogenase flux increases to cope with the extra NADH demand due to alternative oxidase while the opposite occurs in the presence of the ectopic NADH kinase. Experimental data constrained in silico analysis of cytosolic NADH kinase overexpression impact on internal metabolic fluxes (Learn phase) In silico simulations of the Rol-producing strain co-expressing the cytosolic S. cerevisiae NADH kinase (cPOS5) gene revealed trends that closely matched previously reported experimental observations (35). First, our genome-scale based simulation predicted just a negligible increase in ATP regeneration rate when the cPOS5 kinase is expressed. This is coherent with the previous study, where ATP/ADP ratio in the cPOS5 strain remained unaltered, indicating that ATP homeostasis is not substantially perturbed (35). Second, consistent with the remarkable increase in the NADPH/NADP ratio observed in (35), our simulations showed a clear increase in the NADPH regeneration capacity, supporting the notion that cPos5 activity effectively redirects reducing equivalents towards the NADPH pool. Finally, both the computational predictions and cultivation data provided in (35) demonstrated a reduction in biomass yield, which can be attributed to the ATP‑dependent phosphorylation of NADH and the resulting partial dissipation of cellular energy. Together, these concordant results reinforce the mechanistic link between cofactor redistribution caused by cPOS5 kinase and the metabolic adjustments observed in vivo . Experimental data constrained in silico analysis of alternative oxidase overexpression impact on internal metabolic fluxes (Learn phase) The in silico analysis of the HcAtOX-overexpressing strain reveals several metabolic trends that closely parallel observations reported for S. cerevisiae strains ectopically expressing the same alternative oxidase. First, the presence of AOX consistently increased glucose and oxygen turnover rates, indicating enhanced glucose oxidation, a phenomenon also evident from cultivation data in both previous studies. This metabolic shift is accompanied by a reduction in biomass yield, a recurring outcome associated with AOX overexpression. Our model further predicts an increased flux through isocitrate dehydrogenase, aligning with the elevated enzymatic activity assays reported in (36). Similarly, the enhanced mitochondrial regeneration capacity of NADH and NAD + predicted in our simulations mirrors the higher intracellular levels of these cofactors measured experimentally in that study. HcAtOX expression also exerts a pronounced influence on mitochondrial metabolism: Most TCA cycle reactions show increased fluxes while glyoxylate cycle fluxes decreases, matching the transcriptional up-regulation of TCA genes and down-regulation of glyoxylate cycle genes described in (36). Finally, although AOX provides an additional respiratory route by uncoupling NADH oxidation from proton translocation—thereby dissipating energy and contributing to lower biomass yields—its impact on ATP metabolism appears context-dependent. Our simulations predict a hindered ATP turnover rate, whereas previous studies reported an increased P/O ratio and reduced ATP requirement for biomass formation in S. cerevisiae AOX strains (35), suggesting that HcAtOX-driven metabolic rewiring may enhance ATP synthesis efficiency under certain conditions. Ultimately, HcAtOX overexpression seems to mitigate well known overflow-metabolism side-effects in S. cerevisiae growing on glucose. However, this property does not provide a functional advantage in our case, as high glucose concentrations would repress the AOX promoter driving recombinant protein production. On the contrary, this suggests that HcAtOX ectopic overexpression is more likely to impose a metabolic burden on K. phaffii under the tested growth conditions. Experimental data constrained in silico analysis of co-overexpression of both ectopic genes impact on internal metabolic fluxes (Learn phase) The simulation of the double‑mutant strain co-expressing cPOS5 and the alternative oxidase (HcAtOX) genes revealed an unexpectedly complex metabolic behavior, as the two ectopic activities appear to perturb the network in opposing directions. While cPOS5 drives NADH consumption and enhances NADPH formation, AOX provides an additional sink for NADH oxidation, and the simultaneous presence of both functions generates flux patterns that are not straightforward to interpret. Despite this apparent antagonism, the dominant features of the flux distribution can be attributed to a pronounced increase in glucose uptake, which supplies elevated levels of both NADH and ATP through glycolysis. This enhanced glycolytic contribution could explain the observed decrease in the proportion of methanol channeled through the dissimilatory pathway, as the cell becomes less dependent on methanol‑derived NADH for energy generation. Conclusions In this study, we engineered K. phaffii ’s redox metabolism by integrating ectopic genes encoding enzymes that convert NADH into NADPH or NAD⁺, aiming to balance the redox equivalents demands required for recombinant protein synthesis and secretion. We demonstrate that heterologous expression of c POS5 and HcAtOX, encoding an NADH kinase targeted to the cytosol and an alternative NADH oxidase, respectively, can enhance Rol production under methanolic growth conditions. Chemostat cultivations revealed increases in Rol titers ranging among 35–50% when c POS5 was overexpressed, either alone or in combination with HcAtOX. Our results show that the expression level of ectopic genes is a critical determinant of their metabolic impact, influencing both redox cofactor availability and global flux redistribution. Moreover, combining different candidate genetic modifications can lead to additive or even synergistic effects beneficial for recombinant protein production. The in silico initial exploration indicated that metabolic and production phenotypes depend strongly on the carbon source(s) supplied. These findings suggest that the optimal genetic modification strategy is growth condition-dependent; therefore, for each envisaged or constrained bioprocess strategy/condition, a different engineering approach should be identified, e.g. by following the workflow employed in this study. Additional in silico simulations using the genome-scale metabolic model informed with the experimental data obtained from cultivations of the engineered strains revealed metabolic perturbations widespread the whole metabolic network. The agreement between predicted flux changes and available transcriptomic data further supports the model’s accuracy and highlights its value in the DBTL cycle. Beyond recombinant protein production, the redox engineering strategy explored here can be readily extended to other biotechnological applications in K. phaffii , particularly in pathways where redox cofactor unbalance becomes a metabolic bottleneck. Nevertheless, the genetic robustness of the redox-engineered strains should be evaluated in long-term cultivations, as redox/energy status may affect cell’s lifespan. Finally, assessing the applicability of the workflow developed in this study across different substrates, cultivation modes, and target products will further validate its potential. Declarations Conflicts of interest : No potential conflict of interest is reported by the authors. Funding: This study was supported by the projects CTQ2016-74959-R, PID2019-104666GB-I00, and PID2022-136936OB-I00 of the Spanish Ministry of Economy, Industry, and Competitiveness. Sergi Monforte was recipient of the PhD fellowship FPI BES-2014-067935 of the Spanish Ministry of Science and Innovation. Eric Anton is recipient of the PhD fellowship FI (2022FI_B_00639) of the AGAUR (Catalan Government. The research group is supported grants 2021-SGR-00143 from the AGAUR. Author Contribution EA performed in silico simulations, constructed and assessed the strain co-overexpressing both the NADH kinase and alternative oxidase encoding genes, and contributed to the manuscript drafting. SM built the strains overexpressing an alternative oxidase encoding gene, performed the cultures of these strains, as well as contributing to the manuscript drafting. AQ built the strains overexpressing a NADH kinase encoding gene and assisted in the screening of these strains. JA contributed to the design of the in silico experiments and interpretation of the in silico and experimental results and reviewed the manuscript. PF participated in the conceptual design of the experiments, interpretation of the experimental results and contributed to the manuscript drafting. Data Availability All data supporting the findings of this study are available within the paper and its Supplementary Information. Raw data will be made available on request. References Glick BR. Metabolic load and heterologous gene expression. Biotechnology Advances. 1995. p. 247–61. doi:10.1016/0734-9750(95)00004-A PubMed PMID: 14537822. Ferrer P, Albiol J. 13 C-based metabolic flux analysis of recombinant Pichia pastoris . Methods Mol Biol. 2014;1191:291–313. doi:10.1007/978-1-4939-1170-7_17 PubMed PMID: 25178797. Zahrl RJ, Gasser B, Mattanovich D, Ferrer P. 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CRISPR/Cas9-mediated homology-directed genome editing in Pichia pastoris . In: Methods in Molecular Biology. Humana Press Inc.; 2019. p. 211–25. doi:10.1007/978-1-4939-9024-5_9 PubMed PMID: 30737742. Jordà J, Jouhten P, Cámara E, Maaheimo H, Albiol J, Ferrer P. Metabolic flux profiling of recombinant protein secreting Pichia pastoris growing on glucose:methanol mixtures. Microb Cell Fact. 2012;11. doi:10.1186/1475-2859-11-57 Guillén M, Benaiges MD, Valero F. Comparison of the biochemical properties of a recombinant lipase extract from Rhizopus oryzae expressed in Pichia pastoris with a native extract. Biochem Eng J. 2011 Apr 15;54(2):117–23. doi:10.1016/j.bej.2011.02.008 Tomàs-Gamisans M, Ferrer P, Albiol J. Fine-tuning the P. pastoris iMT1026 genome-scale metabolic model for improved prediction of growth on methanol or glycerol as sole carbon sources. Microb Biotechnol. 2018;11(1):224–37. doi:10.1111/1751-7915.12871 Ebrahim A, Lerman JA, Palsson BO, Hyduke DR. COBRApy: COnstraints-Based Reconstruction and Analysis for Python. BMC Syst Biol. 2013 Aug 8;7. doi:10.1186/1752-0509-7-74. Zhang J, Sonnenschein N, Pihl TPB, Pedersen KR, Jensen MK, Keasling JD. Engineering an NADPH/NADP + Redox Biosensor in Yeast. ACS Synth Biol. 2016 Dec 16;5(12):1546–56. doi:10.1021/acssynbio.6b00135. Chung BKS, Lee DY. Flux-sum analysis: A metabolite-centric approach for understanding the metabolic network. BMC Syst Biol. 2009 Dec 19;3. doi:10.1186/1752-0509-3-117. Monforte Mercado S. Systems metabolic engineering for recombinant protein production in Pichia pastoris . 2019. Universitat Autònoma de Barcelona. https://hdl.handle.net/10803/669385 Tomàs-Gamisans C, Andrade M, Maresca C, Monforte F, Ferrer S, Albiol P. Redox Engineering by Ectopic Overexpression of NADH Kinase in Recombinant Pichia pastoris ( Komagataella phaffii ): Impact on cell physiology and recombinant production of secreted proteins. 2020. doi:10.1128/AEM Niu H, Jost L, Pirlot N, Sassi H, Daukandt M, Rodriguez C, et al. A quantitative study of methanol/sorbitol co-feeding process of a Pichia pastoris Mut+ /pAOX1-lacZ strain. Microb. Cell Fact. 12, 33 (2013). https://doi.org/10.1186/1475-2859-12-33 Jordà Murria J. Anàlisi quantitatiu de l’efecte de l’expressió d’una lipasa recombinant sobre el metabolisme central de Pichia pastoris . 2013. Universitat Autònoma de Barcelona. https://hdl.handle.net/10803/117310 Bertels LK, Murillo LF, Heinisch JJ. The pentose phosphate pathway in yeasts–more than a poor cousin of glycolysis. Biomolecules. 2021. doi:10.3390/biom11050725. Hou J, Lages NF, Oldiges M, Vemuri GN. Metabolic impact of redox cofactor perturbations in Saccharomyces cerevisiae. Metab Eng. 2009 Jul;11(4–5):253–61. doi:10.1016/j.ymben.2009.05.001. Vemuri GN, Eiteman MA, Mcewen JE, Olsson L, Nielsen J. Increasing NADH oxidation reduces overflow metabolism in Saccharomyces cerevisiae . Proc Natl Acad Sci U S A. 2007 Feb 13;104(7):2402-7. doi: 10.1073/pnas.0607469104. Additional Declarations No competing interests reported. Supplementary Files SupplementaryFilesEAntonetal.docx Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 05 May, 2026 Reviewers agreed at journal 04 May, 2026 Reviewers agreed at journal 28 Apr, 2026 Reviewers invited by journal 28 Apr, 2026 Editor assigned by journal 24 Apr, 2026 Submission checks completed at journal 24 Apr, 2026 First submitted to journal 17 Apr, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9453242","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":634571027,"identity":"1bb0323f-0a8b-45b1-9904-a3d0189fb9b7","order_by":0,"name":"Eric Antón","email":"","orcid":"","institution":"Autonomous University of Barcelona","correspondingAuthor":false,"prefix":"","firstName":"Eric","middleName":"","lastName":"Antón","suffix":""},{"id":634571028,"identity":"9959c436-a4d0-479b-88cc-6658f9552686","order_by":1,"name":"Sergi Monforte","email":"","orcid":"","institution":"Autonomous University of Barcelona","correspondingAuthor":false,"prefix":"","firstName":"Sergi","middleName":"","lastName":"Monforte","suffix":""},{"id":634571029,"identity":"c1673f28-5215-4647-9c4e-35eab0e2a80e","order_by":2,"name":"Ane Quesada-Ganuza","email":"","orcid":"","institution":"Autonomous University of Barcelona","correspondingAuthor":false,"prefix":"","firstName":"Ane","middleName":"","lastName":"Quesada-Ganuza","suffix":""},{"id":634571030,"identity":"d8b4bf58-3c4c-49b9-9fde-27ca60e19a8c","order_by":3,"name":"Joan Albiol","email":"","orcid":"","institution":"Autonomous University of Barcelona","correspondingAuthor":false,"prefix":"","firstName":"Joan","middleName":"","lastName":"Albiol","suffix":""},{"id":634571031,"identity":"2d5b5e96-6244-4811-97a0-2561f84d2870","order_by":4,"name":"Pau Ferrer","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1ElEQVRIiWNgGAWjYLCCCjDJfICBsYFYLWfAJFsCRAsb8Vp4DIjTYs5+xoDhQMW9fH7pM98kfu5gkOefT8B1lj05QC1nii1n9uVuk+w9w2A44xgBWwwO5Bgwf2xLMDA4w7tNgreNIYGBoJbzbwwYDv5LMLA/w/NM8i9QizxBLTeADjvYALSFh4dNGmSLASEtljOeFRw4cCzBQOIMm7G17BkJw43HEvBrMedP3vjgQE2CAX8P88Obb3fYyMsdPkDAYUCMrESCgKugWkbBKBgFo2AU4AcAvIRBdIV1htUAAAAASUVORK5CYII=","orcid":"","institution":"Autonomous University of Barcelona","correspondingAuthor":true,"prefix":"","firstName":"Pau","middleName":"","lastName":"Ferrer","suffix":""}],"badges":[],"createdAt":"2026-04-18 00:53:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9453242/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9453242/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":108804516,"identity":"c9e9ebb8-1bfc-4b61-96d0-1544fd3afd47","added_by":"auto","created_at":"2026-05-08 15:21:10","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":121787,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eHeat map representation of the \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein silico \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003epredicted impact of gradually increasing the ectopic genes expression on macroscopic physiological parameters. \u003c/strong\u003eThe simulation results depicted are the ones corresponding alternative oxidase (A, B) or NADH kinase (C, D) expression on either Glucose/Methanol (A, C) or Glycerol/Methanol (B, D) 60%:40% w/w culture conditions. (*) The flux variation rate color scale for Rol production ranges from 0.5 to 1.5.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-9453242/v1/7ae60a651167e2191ca3a5cd.png"},{"id":108805982,"identity":"e517f8d8-b8a1-4768-8851-0ea79f5b5ea2","added_by":"auto","created_at":"2026-05-08 15:27:22","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":122438,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eHeat map representation of the \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein silico \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003epredicted impact of gradually increasing the ectopic genes expression on redox cofactors regeneration rate. \u003c/strong\u003eThe simulation results depicted are the ones corresponding alternative oxidase (A, B) or NADH kinase (C, D) expression on either Glucose/Methanol (A, C) or Glycerol/Methanol (B, D) 60%:40% (w/w) culture conditions.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-9453242/v1/751b95d09e00158c31a2f914.png"},{"id":108804503,"identity":"e481519b-7d4c-465b-9c41-f04a02d90d2f","added_by":"auto","created_at":"2026-05-08 15:20:56","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":37801,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSmall\u003c/strong\u003e‑\u003cstrong\u003escale characterization of Rol\u003c/strong\u003e‑\u003cstrong\u003eproducing strains overexpressing HcAtOX and/or cPOS5.\u003c/strong\u003e Specific lipase activity values are normalized to the reference strain (X‑33 Rol). Clones are grouped by quartiles (boxes), with whiskers representing variability outside these ranges. Dot color indicates the cPOS5 gene copy number, as determined by ddPCR: \u003cstrong\u003eblack\u003c/strong\u003e for one copy, \u003cstrong\u003ered\u003c/strong\u003e for two copies, and \u003cstrong\u003ewhite\u003c/strong\u003e for more than two copies. cPOS5 gene copy number was determined through ddPCR.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-9453242/v1/85fe655ab74acc8bb428193e.png"},{"id":108805505,"identity":"a093c584-bb55-4e3d-8af0-284677a20dac","added_by":"auto","created_at":"2026-05-08 15:26:08","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":205023,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSimplified \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eP. pastoris \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003emetabolic network depicting flux variations caused by ectopic genes overexpression\u003c/strong\u003e \u003cstrong\u003eA.\u003c/strong\u003e Mock network \u003cstrong\u003eB.\u003c/strong\u003e 1 HcAtOX copy \u003cstrong\u003eC.\u003c/strong\u003e 2 cPOS5 copies \u003cstrong\u003eD\u003c/strong\u003e. 2 cPOS5 copies + 1 HcAtOX copy (POSNOX). Reactions showing increased or decreased flux values are represented by red and blue lines respectively, whereas those not affected reactions are coloured in grey. Black diamonds in the mock network mark native reactions involved in redox cofactor production and/or consumption for which transcriptomic data previously obtained under the same growth conditions (30). Specifically, these reactions are glyceraldehyde-3-phosphate dehydrogenase \u003cstrong\u003e(1)\u003c/strong\u003e, glucose-6-phosphate dehydrogenase \u003cstrong\u003e(2)\u003c/strong\u003e and isocitrate dehydrogenase (NAD\u003csup\u003e+\u003c/sup\u003e-dependent) \u003cstrong\u003e(3)\u003c/strong\u003e. Transcriptomic changes are represented with either arrows or boxes containing a “T” as follows: decreased gene expression is shown with a blue downward arrow, increased expression with a red upward arrow, and grey boxes indicate genes for which no significant change in expression was detected.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-9453242/v1/c3a564dc8b57660cca8d515e.png"},{"id":108809567,"identity":"b047074c-6536-4a21-a2d8-1f4c1303b3d7","added_by":"auto","created_at":"2026-05-08 15:53:53","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":956044,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9453242/v1/c806fc15-563a-4ecc-ab3c-00445ef65f51.pdf"},{"id":108588010,"identity":"a16cf377-1728-460e-8596-5f93872d6796","added_by":"auto","created_at":"2026-05-06 09:14:49","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":242220,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFilesEAntonetal.docx","url":"https://assets-eu.researchsquare.com/files/rs-9453242/v1/7497e40449cd21e98264c1ae.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Metabolic impact of redox perturbations on recombinant protein production in Komagataella phaffii under methanolic growth conditions","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOverproduction of recombinant proteins in yeast such as \u003cem\u003eKomagataella phaffii\u003c/em\u003e (also known as \u003cem\u003ePichia pastoris\u003c/em\u003e), exerts a significant metabolic burden to the cell due to the drain of resources from cell maintenance and growth (1,2). This contributes to the metabolic stress caused by overexpression of recombinant proteins, negatively impacting productivity (2,3). Such metabolic burden may be triggered by several factors: \u003cem\u003ede novo\u003c/em\u003e synthesis of energetically costly amino acids, overloaded protein folding machinery, posttranslational protein modifications and secretion processes. These lead to energetic and redox balance alterations and a suboptimal cell physiological state to support recombinant protein production. Specifically, production of recombinant proteins in \u003cem\u003eK. phaffii\u003c/em\u003e leads to a redistribution of metabolic fluxes to increase supply of ATP and NADH (4\u0026ndash;7). In addition, several \u003csup\u003e13\u003c/sup\u003eC-MFA studies in recombinant \u003cem\u003eK. phaffii\u003c/em\u003e point at increased NADPH supply through pentose phosphate pathway (PPP) in production strains (8,9).\u003c/p\u003e \u003cp\u003eSeveral metabolic engineering strategies targeting redox metabolism have been explored to overcome these limitations in \u003cem\u003eK. phaffii\u003c/em\u003e (3). For instance, overexpression of genes encoding PPP enzymes increased NADPH supply, allowing for improved recombinant protein yield under glucose-growing conditions, using the constitutive GAP promoter (P\u003csub\u003e\u003cem\u003eGAP\u003c/em\u003e\u003c/sub\u003e) expression system (10). Similarly, we have demonstrated the positive effect of increasing NADPH availability by overexpressing a heterologous NADH kinase encoding \u003cem\u003ePOS5\u003c/em\u003e gene from the yeast \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e (11) in a \u003cem\u003eK. phaffii\u003c/em\u003e strain producing an antibody fragment under the control of P\u003csub\u003e\u003cem\u003eGAP\u003c/em\u003e\u003c/sub\u003e, thereby increasing secreted product titers by 1.55 and 2-fold in normoxic and hypoxic glucose-limited chemostats, respectively. Notably, the use of glycerol as an alternative C-source resulted in a lower effect \u003cem\u003e(12)\u003c/em\u003e. Overall, these studies strongly support the hypothesis of a higher limitation of recombinant protein production by NADPH when cells are grown on glucose, as well as pointing out that different redox, NADPH, engineering solutions may be needed depending on the growth conditions, as expected from the observation that sources of NADPH may vary depending on the C-source (13). This may be particularly relevant when the use of alternative C-sources such as methanol (or combinations thereof) is foreseen.\u003c/p\u003e \u003cp\u003eBesides, the impact of NADH availability on \u003cem\u003eK. phaffii\u003c/em\u003e growth and protein production has also been explored by overexpressing a heterologous NADH oxidase from \u003cem\u003eLactococcus lactis\u003c/em\u003e (14). By oxidizing NADH to NAD\u003csup\u003e+\u003c/sup\u003e, the NADH/NAD\u003csup\u003e+\u003c/sup\u003e ratio increase caused by recombinant protein production was counteracted. This resulted in an upregulation of methanol metabolism genes and recombinant CALB lipase gene (expressed under the control of the methanol-inducible \u003cem\u003eAOX1\u003c/em\u003e promoter), which in turn allowed to improve methanol uptake rate and recombinant protein production. Moreover, by further co-overexpressing an adenylate kinase (\u003cem\u003eADK1\u003c/em\u003e) from \u003cem\u003eS. cerevisiae\u003c/em\u003e, the ATP supply in this strain, which was reduced by the lower NADH availability for ATP synthesis, was restored, achieving even higher production levels.\u003c/p\u003e \u003cp\u003eIn the light of these studies and in order to develop a lean workflow to systematically assess the impact of different redox engineering strategies on NADPH regeneration and heterologous protein production in \u003cem\u003eK. phaffii\u003c/em\u003e under methanolic growth conditions, a model-based Design-Build-Test-Learn (DBTL) cycle approach, enabled by \u003csup\u003e13\u003c/sup\u003eC-fluxomics, has been conceived and benchmarked with two different genetic targets and an exemplary well-defined growth condition. To this end, in this study, we have further investigated the effect of cofactor regeneration on the heterologous production of a \u003cem\u003eRhizopus oryzae\u003c/em\u003e lipase (Rol) and the central carbon metabolism of \u003cem\u003eK. phaffii\u003c/em\u003e growing under methanolic conditions, by overexpressing a heterologous NADH kinase encoding gene \u003cem\u003ePOS5\u003c/em\u003e from \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e and an alternative oxidase (\u003cem\u003eAtOX\u003c/em\u003e) from \u003cem\u003eHistoplasma capsulatum\u003c/em\u003e. The NADH kinase was targeted to the cytosol, providing an extra NADPH source, whereas the alternative oxidase is an enzyme located in the inner mitochondrial membrane that provides an alternative route to transfer electrons from the ubiquinol pool to oxygen, bypassing several proton-pumping steps this way. Similarly, this alternative oxidase has been shown to locate in mitochondria when ectopically overexpressed in \u003cem\u003eS. cerevisiae\u003c/em\u003e (15). A previous study showed an important physiological impact when the endogenous alternative oxidase gene in \u003cem\u003eK. phaffii\u003c/em\u003e was either overexpressed or eliminated (16), but its effect in recombinant protein production was not investigated.\u003c/p\u003e \u003cp\u003eFirstly, genome scale model-based simulations were performed to assess the impact of increased flux through the ectopic reactions on both macroscopic growth parameters, NADPH regeneration and product formation under a defined set of growth conditions (mixed multicarbon source:methanol chemostat cultivations), for which previous \u003csup\u003e13\u003c/sup\u003eC-Metabolic Flux analyses (\u003csup\u003e13\u003c/sup\u003eC-MFA) were available (7,17,18). Second, the redox-engineered strains were constructed and were tested at shake flask scale and then further characterized in 1-L scale chemostat cultures. Finally, to gain a deeper insight into the physiological effect of these modifications, an \u003cem\u003ein silico\u003c/em\u003e interpretation of the metabolic effect of these genetic modifications was also performed.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStrains and plasmids\u003c/h2\u003e \u003cp\u003eA \u003cem\u003eK. phaffii\u003c/em\u003e X-33-derived strain containing a single copy of the gene encoding the lipase of \u003cem\u003eRhizopus oryzae\u003c/em\u003e (\u003cem\u003eROL\u003c/em\u003e) was used in this study (19). The expression cassette of this gene, constructed using the pPICZαA vector (Invitrogen-Thermo Fisher Scientific, California, USA), is under control of the methanol-inducible \u003cem\u003eAOX1\u003c/em\u003e promoter (P\u003csub\u003e\u003cem\u003eAOX1\u003c/em\u003e\u003c/sub\u003e) and includes the encoding region of the \u003cem\u003eS. cerevisiae\u003c/em\u003e α-mating factor secretion signal (20).\u003c/p\u003e \u003cp\u003eThe genes encoding the NADH kinase Pos5p (\u003cem\u003ePOS5\u003c/em\u003e) from the yeast \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e and the alternative oxidase (\u003cem\u003eAtOX\u003c/em\u003e) from the fungi \u003cem\u003eHistoplasma capsulatum\u003c/em\u003e were codon-optimized for \u003cem\u003eK. phaffii\u003c/em\u003e and synthetized by Geneart (ThermoFischer Scientific) and cloned into a pPUZZLE vector (21) under control of GAP promoter. In addition, the first 49 bp of \u003cem\u003ePOS5\u003c/em\u003e ORF, encoding region of the mitochondrial signal peptide, were removed, i.e. ensuring both cytosolic and mitochondrial location of the corresponding protein product, and cloned in the pPUZZLE vector. Construction of \u003cem\u003ePOS5\u003c/em\u003e and truncated \u003cem\u003ePOS5\u003c/em\u003e expression vectors have been previously described in detail (12). Briefly, 2 vectors were constructed in this study: pPUZZLE_cPOS5 and pPUZZLE_\u003cem\u003eHc\u003c/em\u003eAOX1 (\u003cb\u003eSupplementary figure I\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eThe single mutants were generated by genomic integration of the c\u003cem\u003ePOS5, mPOS5\u003c/em\u003e and \u003cem\u003eHc\u003c/em\u003eAOX1 \u003cem\u003eexpression vectors\u003c/em\u003e. pPUZZLE plasmids were linearized using AvrII (New England Biolabs, Massachusetts, USA) and introduced into \u003cem\u003eK. phaffii\u003c/em\u003e X-33/ROL using 100 ng of DNA. Transformation was performed by electroporation with a Gene PulserXcell\u0026trade; Electroporation System (Bio-Rad, Hercules, CA, USA) using as parameters: 1550 V, 25 \u0026micro;F and 200 Ω. Positives clones were selected by growing on Yeast Peptone Dextrose (YPD) plates with 500 \u0026micro;g/mL of G418 (Geneticin) and checked by colony PCR (all primers used in this study are described in \u003cb\u003eSupplementary table I\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eThe combined POS5 and \u003cem\u003eHc\u003c/em\u003eAtOX1 mutants (POSNOX) were obtained by transforming the strain harboring the alternative oxidase with 500 ng of linearized with AvrII BB3eH_14_pGAP_cPOS5_ScCYCtt plasmid (22). 200 ng/mL of Hygromycin was added on YPD plates as selection marker. Given that clonal variation was expected in the number of cPOS5 cassette copies integrated in the genome, around 20 positive clones were isolated and characterized. Electrocompetent \u003cem\u003eK. phaffii\u003c/em\u003e X-33 cells were prepared according to established protocols (23). Strains constructed in this study are summarized in Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eList of strains constructed and used in this study\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStrain\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eParental strain\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePlasmids and cassettes used for transformation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGenotype\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAntibiotic resistance\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eReference\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eX-33 ROL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eX-33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epPICZa_ROL plasmid linearized with PmeI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRol-producing strain\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eZeocin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eX-33 ROL cPOS5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eX-33/ ROL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epPUZZLE_cPOS5 linearized with AvrII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRol-producing strain harbouring 1 or more c\u003cem\u003ePOS5\u003c/em\u003e (cytosolic \u003cem\u003ePOS5\u003c/em\u003e) copies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eG418 (Geneticin) and Zeocin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eX-33 ROL HcAtOX\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eX-33 ROL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epPUZZLE_cPOS5 Linearized with AvrII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRol producing strain harbouring 1 or more cytosolic \u003cem\u003eHc\u003c/em\u003eAOX1 copies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eG418 (Geneticin) and Zeocin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eX-33 ROL POSNOX\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eX-33 HcAtOX\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBB3eH_14_pGAP_cPOS5_ScCYCtt Linearized with AvrII\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRol-producing strain harbouring 1 or more c\u003cem\u003ePOS5\u003c/em\u003e copies and 1 HcAOX1 copy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eG418 (Geneticin), Zeocin and Hygromycin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eThis work\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eScreening conditions\u003c/h3\u003e\n\u003cp\u003eTo consider clonal variation of \u003cem\u003eK. phaffii\u003c/em\u003e transformants, five colonies from each strain were selected and further tested in shake flask cultures in triplicate at 25\u0026ordm;C and 150 rpm, as described by C\u0026aacute;mara and coworkers (19). Briefly, strains were first grown in 50 mL of Buffered Minimal Glycerol (BMG) medium in 1-L shake flasks overnight at 25\u0026ordm;C and 150 rpm. Afterwards, reinoculation from BMG into 25 mL of fresh Buffered Minimal Methanol (BMM) medium in 250-mL shake flasks was performed at an initial optical density of 1 and further grown under the same conditions. One pulse of 125 \u0026micro;L of pure methanol was added (0.5% v/v) after 24 h and 48 h of cultivation. Samples for biomass and recombinant protein quantification were taken after 70 h of cultivation.\u003c/p\u003e\n\u003ch3\u003eChemostat cultivation conditions\u003c/h3\u003e\n\u003cp\u003eSelected strains were characterized in duplicate chemostat cultures using a mixed carbon source consisting of 60%:40% glucose:methanol (w/w). Cultivations were performed at a 700-mL working volume in a DASGIP Parallel Bioreactor System (Eppendorf, Germany). Operating conditions were set at 25\u0026deg;C, pH 5.0 controlled through the addition of 15% ammonia (v/v), 700 rpm, 1 vvm inlet gas flow, and a dilution rate (D) of 0.1 h⁻\u0026sup1;. Steady states were monitored along the 4th, 5th, and 6th residence times to ensure stable biomass concentration, CER, OUR, lipase activity and extracellular metabolite profiles. Measurements at the end of the 5th residence time were considered as experimental steady-state growth parameters.\u003c/p\u003e \u003cp\u003eBatch medium contained: 19.95 g/L glycerol, 0.9 g/L citric acid, 6.3 g/L (NH\u003csub\u003e4\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e, 0.01 g/L CaCl\u003csub\u003e2\u003c/sub\u003e, 0.45 g/L KCl, 0.25 g/L MgSO\u003csub\u003e4\u003c/sub\u003e\u0026middot;7H\u003csub\u003e2\u003c/sub\u003eO, 1 mL/L Biotin (0.2 g/L; Sigma, Munich, Germany), 0.3 mL antifoam (Glanapon, Bussetti \u0026amp; Co GmbH, Wien, Austria) and 2.3 mL/L PTM1 trace salts stock solution. Chemostat medium contained: 25g/L total carbon source, 0.42 g/L citric acid, 2.18 g/L (NH\u003csub\u003e4\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e, 5.5 mg/L CaCl\u003csub\u003e2\u003c/sub\u003e\u0026middot;2H\u003csub\u003e2\u003c/sub\u003eO, 0.85 g/L KCl, 0.32 g/L MgSO\u003csub\u003e4\u003c/sub\u003e\u0026middot;7H\u003csub\u003e2\u003c/sub\u003eO, 0.5 mL Biotin (0.2 g/L), 50 \u0026micro;L antifoam (Glanapon) and 0.8 mL PTM1 trace salts stock solution. Trace salt composition was: 6.0 g/L CuSO\u003csub\u003e4\u003c/sub\u003e.5H\u003csub\u003e2\u003c/sub\u003eO; 0.08 g/LNaI; 3.36 g/L MnSO\u003csub\u003e4\u003c/sub\u003e.H\u003csub\u003e2\u003c/sub\u003eO; 0.2 g/LNa\u003csub\u003e2\u003c/sub\u003eMoO\u003csub\u003e4\u003c/sub\u003e.2H\u003csub\u003e2\u003c/sub\u003eO; 0.02 g/L H\u003csub\u003e3\u003c/sub\u003eBO\u003csub\u003e3\u003c/sub\u003e; 0.82 g/L CoCl\u003csub\u003e2\u003c/sub\u003e.6H\u003csub\u003e2\u003c/sub\u003eO; 20 g/L ZnCl\u003csub\u003e2\u003c/sub\u003e; 65 g/L FeSO\u003csub\u003e4\u003c/sub\u003e.7H\u003csub\u003e2\u003c/sub\u003eO and 5.0 mL H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e (95\u0026ndash;98% w/w). Medium pH was adjusted to 5 with 25% HCl.\u003c/p\u003e \u003cp\u003eAll chemostat cultivations were performed at least in duplicate.\u003c/p\u003e\n\u003ch3\u003eAnalytical methods\u003c/h3\u003e\n\u003cp\u003eBiomass determination\u003c/p\u003e \u003cp\u003eBiomass optical density was measured at 600 nm using a DR3900 spectrophotometer (Hach, Bizkaia, Spain). Dry cell weight (DCW) was determined using pre-weighed dried glass fiber filters (Millipore, Massachusetts, USA) as reported by Jord\u0026agrave; and coworkers (24). Both measurements were performed in triplicate.\u003c/p\u003e \u003cp\u003eLipolytic activity assay\u003c/p\u003e \u003cp\u003eLipase activity in the cleared cultivation broth was quantified using a colorimetric assay based on the hydrolysis of p-nitrophenyl butyrate as substrate, following the method described by Guill\u0026eacute;n et al. (2011)(25).\u003c/p\u003e \u003cp\u003eMetabolite quantification\u003c/p\u003e \u003cp\u003eResidual glucose and methanol concentrations in the cleared cultivation broth were measured by means of HPLC. Specifically, a Vanquish Flex UHPLC system (Thermo Fisher Scientific, Waltham, MA, USA) equipped with an ICSep ICE-COREGEL 87H3 (Transgenomic) ion exchange column and a Water 2410 (Waters) refraction index detector. CROMELEON software (Dionex) was used for data analysis.\u003c/p\u003e \u003cp\u003e \u003cb\u003eHeterologous\u003c/b\u003e \u003cb\u003ePOS5\u003c/b\u003e \u003cb\u003egene copy number determination\u003c/b\u003e\u003c/p\u003e \u003cp\u003eTo quantify the heterologous (from \u003cem\u003eS. cerevisiae\u003c/em\u003e) \u003cem\u003ePOS5\u003c/em\u003e gene dosage, ddPCR analysis was performed. First, genomic DNA from POSNOX clones was extracted using a commercial kit (Wizard Genomic DNA Purification Kit, Promega, Madison, WI) following the manufacturer\u0026rsquo;s instructions and quantified in a NanoDrop 2000 spectrophotometer (Thermo Scientific). QX200 ddPCR EvaGreen Supermix reaction cocktail (Bio-Rad) was used to carry out the assay. Operational parameters can be consulted in the manufacter\u0026rsquo;s guideline for this product. Once reaction mixtures were prepared a QX200 Droplet Generator (Bio-Rad) was employed to generate droplets and the samples were transferred into a 96-well plate. After thermal cycling procedure, the plate was placed in a QX200 Droplet Reader (Bio-Rad). Results data acquisition was conducted with the software QuantaSoft v.1.7. The \u003cem\u003ePOS5\u003c/em\u003e gene copy number was determined by calculating the ratio of positive droplets between the sample and a reference sample carrying one copy of \u003cem\u003ePOS5\u003c/em\u003e.\u003c/p\u003e \u003cp\u003ePrior temperature and DNA concentration gradient ddPCRs were performed to determine the optimal conditions for the reaction and the one POS5 Copy reference clone. For these tests a one copy housekeeping gene, namely β-Actin (ACT1) was selected to normalize data. The annealing temperature and the DNA quantity per sample were set to 58\u0026deg;C and 1 ng (or 0.05 ng/\u0026micro;L) respectively.\u003c/p\u003e \u003cp\u003ePrimers used for this assay are described in \u003cb\u003eSupplementary table I\u003c/b\u003e.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIn silico\u003c/b\u003e \u003cb\u003emetabolic flux analysis\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe iMT1026 v.3.0 genome scale model of \u003cem\u003eK. phaffii\u003c/em\u003e (26) was used in the COBRApy 0.26.2 (27) toolbox with CPLEX 12.9 as solver. The simulations of the reference Rol-producing strain were done using pFBA, maximizing the biomass reaction and defining a set of reaction ratios extracted from previous \u003csup\u003e13\u003c/sup\u003eC-tracer experiments performed on mixed methanol and glucose or glycerol chemostat cultivations (7,18) as constraints depending on the simulated cultivation conditions (60/40% Glucose/Methanol or 60/40% Glycerol/Methanol, respectively). Simulations of the redox-engineered strains were performed using Non-Linear Minimization of Metabolic Adjustment (MOMA), with the wild-type simulations as reference conditions. For all simulations performed, the carbon source uptake rate was fixed to match the reference X-33 ROL strain cultivations experimental data. In the \u003cem\u003ePOS5\u003c/em\u003e- and HcAtOX-overexpressing strains simulations, the ectopic reactions were added at fixed flux values ranging from 0 to 0.2 mmol/(gDCW\u0026middot;h) to explore its impact in the metabolic network.\u003c/p\u003e \u003cp\u003eRedox cofactors regeneration was calculated by summing the absolute flux value of all the reaction involved in the consumption or generation of a cofactor and diving it by 2 as detailed elsewhere (28). Constrained based models do not include metabolite concentrations as state variables. Thus, redox cofactors ratios cannot be calculated. Therefore, we used metabolite turnover rates (AKA flux-sum (29) as a proxy for changes in redox balance. Metabolites flux-sum were calculated as described in Eq.\u0026nbsp;1:\u003c/p\u003e \u003cp\u003e \u003cimg 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\" style=\"width: 246px;\"\u003e \u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;Eq.\u0026nbsp;1\u003c/p\u003e \u003cp\u003eWhere S\u003csub\u003eij\u003c/sub\u003e is the stoichiometric coefficient of metabolite i in reaction j and v\u003csub\u003ej\u003c/sub\u003e is the flux of reaction j.\u003c/p\u003e\n\u003ch3\u003eStatistical analyses\u003c/h3\u003e\n\u003cp\u003eUnless otherwise stated, data are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation of at least triplicate samples. To test for significance when comparing experimental data, obtained from continuous cultures and transcriptomic experiments, a Student\u0026rsquo;s \u003cem\u003et\u003c/em\u003e-test was performed using the Microsoft Excel software. A p-value lower than 0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"Results and discussion","content":"\u003cp\u003e\u003cstrong\u003eIn silico\u003c/strong\u003e \u003cstrong\u003eprediction of increased Pos5 and Aox1 activity impact on physiological parameters and recombinant protein production under different cultivation conditions (Design phase)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe explored beforehand the potential effects of ectopically overexpressing the \u003cem\u003eS. cerevisiae\u003c/em\u003e\u0026rsquo;s c\u003cem\u003ePOS5\u003c/em\u003e (truncated \u003cem\u003ePOS5\u003c/em\u003e for cytosolic localization) or \u003cem\u003eH. capsulatum\u003c/em\u003e\u0026rsquo;s AOX1 (HcAtOX1) genes on the physiological growth parameters and intracellular fluxes distribution by performing genome-scale model-based simulations in which we increased iteratively the flux value of their corresponding reactions and fixing them as constraints. The resulting metabolic flux distribution was obtained by performing an NL MOMA, using as reference simulation a previous pFBA constrained with reaction ratios obtained from earlier \u003csup\u003e13\u003c/sup\u003eC-MFA studies (7,17,18). We conducted this procedure for both ectopic reactions separately under two different co-substrate feeding regimes under metabolic steady state conditions, that is, glucose or glycerol plus methanol in a fixed ratio (60% co-substrate\u0026thinsp;+\u0026thinsp;40 % methanl, (w/w)).\u003c/p\u003e\n\u003cp\u003eThe simulations showed modest perturbations in the respiratory parameters (specific oxygen consumption and CO\u003csub\u003e2\u003c/sub\u003e production rates, q\u003csub\u003eO2\u003c/sub\u003e and q\u003csub\u003eCO2\u003c/sub\u003e) when flux through the ectopic reactions was increased. Notably, while growth progressively diminished in all cases, production of the recombinant protein Rol increased notably in the POS5 case study (Fig. \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). In contrast, the introduction of HcAOX1 resulted in a negative effect on protein production as well as inducing minor respiratory adjustments. Overall, the simulation results depicted in Fig. \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e suggested that recombinant protein production is primarily impacted by cPOS5 overexpression, while HcAOX1 does not show a significant benefit, with respiration and final biomass remaining essentially unaffected, irrespective of the co-substrate (glucose or glycerol) used.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIn silico\u003c/strong\u003e \u003cstrong\u003eprediction of the impact of Pos5 and AtOX activity on redox cofactors turnover rates (Design phase)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGiven that both ectopic reactions are directly related to the redox pairs NADPH/NADP\u003csup\u003e+\u003c/sup\u003e and NADH/NAD\u003csup\u003e+\u003c/sup\u003e, the NAD(P)H regeneration capacity in the potential engineered strains was evaluated by calculating the turnover rates of these metabolites in the cytosol for every simulation previously performed. In the initial exploration (i.e., scanning ectopic flux from 0 to 0.2 mmol/(gDCW\u0026middot;h)), we observed an increasing tendency in regenerative capacity for all cofactors (Fig. \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), the NADPH/NADP\u003csup\u003e+\u003c/sup\u003e pair showing the most pronounced increases. Remarkably, this pattern was more prominent in the POS5 case, especially when glucose was used as co-substrate, coherent with the fact that Pos5 kinase reaction directly generates NADPH. However, considering that alternative oxidases boost fluxes through the electron transport chain subunits reactions, we expected that the impact on the NAD\u003csup\u003e+\u003c/sup\u003e cofactor regeneration rate would be higher when simulating the AtOX gene overexpression. This outcome could be attributed to the influence that the endogenous mitochondrial Pos5 kinase could have on the energetic state, as it consumes ATP to phosphorylate NADH. Coherently, simulations revealed increasing ATP demand under this scenario (Supplementary Fig. 2). These turnover rate calculations were done for the cytosolic pools of the cofactors, since metabolite pools are differentiated by subcellular location in the genome-scale model. Thereby, we also calculated the turnover rates for the redox cofactors located in the mitochondria (Supplementary Fig. 3), obtaining coherent values. Namely, a higher regeneration rate of mitochondrial NAD\u003csup\u003e+\u003c/sup\u003e and NADH when HcAOX is overexpressed in contrast to ectopic c\u003cem\u003ePOS5\u003c/em\u003e expression. Moreover, the 2cPOS5 and HcAOX1 (POSNOX) combined stran was tested \u003cem\u003ein silico\u003c/em\u003e by fixing the HcAOX1 reaction flux to the estimated value (0.1 mmol/(gDCW\u0026middot;h)) and progressively augmenting the POS5 reaction flux. This analysis revealed an additive effect in the NADPH regeneration capacity (Supplementary Fig. 4).\u003c/p\u003e\n\u003cp\u003eOverall, the initial \u003cem\u003ein silico\u003c/em\u003e exploration revealed that the expression of both ectopic genes would prove beneficial in the redox cofactors\u0026rsquo; regeneration capacity of the engineered strains, thereby motivating subsequent strain construction and testing. Moreover, once the engineered strains characterization in chemostat cultures was completed, simulations were run again using the obtained experimental data as constraints to get new insights on how the metabolic flux could be redistributed because of \u003cem\u003ePOS5\u003c/em\u003e and HcAtOX expression.\u003c/p\u003e\n\u003cp\u003eIn addition, the Glucose 60% / Methanol 40% (w/w) mixture was selected for further characterization of the engineered strains at bioreactor scale based on the following considerations: First, \u003cem\u003ein silico\u003c/em\u003e predictions indicated that recombinant protein production more enhanced when glucose is used as a co‑substrate, compared to the glycerol case. This observation was coherent with previous chemostat cultivation studies using glycerol as co-substrate with the first series of redox engineered Rol-producing strains, where the observed increases in lipase activity relative to the reference strain were not statistically significant (30). Second, perturbations in redox cofactor balance caused by POS5 overexpression, i.e., on the NADPH/NADP\u003csup\u003e+\u003c/sup\u003e ratio, have been shown to be more pronounced when cells are cultivated on glucose compared to glycerol (31). Although using methanol as the sole carbon source was initially considered, literature reports indicate that similar induction levels can be achieved with mixed carbon sources (32). Additionally, methanol appears to function more effectively as an auxiliary energy substrate in \u003cem\u003eK. phaffii\u003c/em\u003e (33).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePOS5 and\u003c/strong\u003e \u003cstrong\u003eHc\u003c/strong\u003e\u003cstrong\u003eAtOX co-expression strain construction (Build)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eElectrocompetent strains carrying a single HcAtOX copy were transformed with a linearized pPuzzle vector containing the cPOS5 kinase. To quantify the number of integrated cPOS5 copies in the resulting transformants, ddPCR analysis was performed on several clones. Most isolates incorporated one, two, or three copies of the NADPH kinase encoding gene (Supplementary table 2), indicating efficient but variable integration of the construct.\u003c/p\u003e\n\u003ch3\u003eSmall scale screening of POS5 and HcAtOX1 co-expressing clones (Test)\u003c/h3\u003e\n\u003cp\u003eAll transformant clones were first cultivated at small scale in BMM medium, followed by assessment of lipase activity to estimate recombinant protein production. Under these screening conditions, all clones transformed with the c\u003cem\u003ePOS5\u003c/em\u003e gene displayed no significant increment in recombinant protein production except for one individual clone harboring 2 copies of the ectopic gene. Interestingly, the majority of HcAtOX clones carrying more than two copies of the cytosolic POS5 kinase displayed a clear increase in lipase activity (Fig. \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). To further evaluate this set of engineered strains, only the clones harboring exactly two POS5 copies were selected for reactor‑scale characterization, as strains with higher copy numbers exhibited a pronounced reduction in growth rate during the inoculum preparation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eReactor-scale characterization of POS5 gene dosage effect and co-overexpression of POS5 and AtOX on recombinant protein production and physiological parameters. (Test)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA strain co‑expressing the ectopic genes investigated in this study was constructed and evaluated in chemostat cultures using 60% (w/w) glucose as a co‑substrate to assess the predicted synergistic effect on protein production. Additionally, a strain harboring a single POS5 kinase copy was examined to verify that one copy of this gene does not enhance Rol synthesis, as observed from small‑scale experiments. Consequently, a new set of cultivations were performed using the reference X-33 ROL strain, HcAtOX, 1cPOS5, 2cPOS5, as well as a strain combining two POS5 copies with one HcAtOX copy.\u003c/p\u003e\n\u003cp\u003eThe 1cPOS5 strain did not enhance Rol production (Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e), confirming that a single POS5 copy is insufficient to improve recombinant protein synthesis. In contrast, the HcAtOX strain exhibited a pronounced physiological impact, including a clear reduction in biomass yield and a hampering of methanol assimilation, as indicated by its lower methanol-specific consumption rate and elevated residual methanol concentrations. This strain also showed a decrease in Rol activity, demonstrating that the introduction of a single HcAtOX copy negatively affects both growth and protein production. Although neither 1cPOS5 nor 2cPOS5 strains substantially altered biomass yield or residual methanol levels, both POS5-based modifications increased respiratory activity, reflected by higher CO₂ secretion rates and elevated RQ values. Strains carrying two POS5 copies (2cPOS5), including the combined 2cPOS5\u0026ndash;HcAtOX construct displayed substantial improvements in Rol production, reaching increases of approximately 30\u0026ndash;40%. Notably, the additional HcAtOX copy in the 2cPOS5-HcAtOX combined mutant did not further enhance Rol synthesis relative to 2cPOS5, but it strongly influenced macroscopic process parameters, including reduced final biomass concentrations and pronounced increases in both oxygen uptake and CO₂ secretion rates.\u003c/p\u003e\n\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1. Macroscopic parameters of the redox engineered and reference Rol-producing strains cultivated in chemostat using a glucose 60% methanol 40% mix as carbon source.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\n \u003cv:shapetype id=\"_x0000_t75\" coordsize=\"21600,21600\" o:spt=\"75\" o:preferrelative=\"t\" path=\"m@4@5l@4@11@9@11@9@5xe\" filled=\"f\" stroked=\"f\"\u003e\u0026nbsp;\u003cv:stroke joinstyle=\"miter\"\u003e\u0026nbsp;\u003cv:formulas\u003e\u0026nbsp;\u003cv:f eqn=\"if lineDrawn pixelLineWidth 0\"\u003e\u0026nbsp;\u003cv:f eqn=\"sum @0 1 0\"\u003e\u0026nbsp;\u003cv:f eqn=\"sum 0 0 @1\"\u003e\u0026nbsp;\u003cv:f eqn=\"prod @2 1 2\"\u003e\u0026nbsp;\u003cv:f eqn=\"prod @3 21600 pixelWidth\"\u003e\u0026nbsp;\u003cv:f eqn=\"prod @3 21600 pixelHeight\"\u003e\u0026nbsp;\u003cv:f eqn=\"sum @0 0 1\"\u003e\u0026nbsp;\u003cv:f eqn=\"prod @6 1 2\"\u003e\u0026nbsp;\u003cv:f eqn=\"prod @7 21600 pixelWidth\"\u003e\u0026nbsp;\u003cv:f eqn=\"sum @8 21600 0\"\u003e\u0026nbsp;\u003cv:f 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\" alt=\"image\"\u003e\n\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental data constrained\u003c/strong\u003e \u003cstrong\u003ein silico\u003c/strong\u003e \u003cstrong\u003eanalysis of ectopic genes overexpression impact on internal metabolic fluxes (Learn phase)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo further understand how the expression of the ectopic genes tweaks \u003cem\u003eK. phaffii\u0026rsquo;s\u003c/em\u003e metabolism, bolstering the regeneration of redox cofactors and, ultimately, enhancing the recombinant protein production, simulations were repeated, setting the carbon source uptake rates obtained in chemostat cultivations as model constraints. The prediction results were visualized in a simplified \u003cem\u003eK. phaffii\u003c/em\u003e metabolic network and further complemented with transcriptomics data of reporter genes previously obtained from analogous cultivations (30). Primarily, there is a redirection of flux to the methanol dissimilatory pathway which takes place regardless of the ectopic gene expressed (Fig. \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). This metabolic shift in the peroxisome is coherent with the increasing NADH regeneration rates seen in the initial \u003cem\u003ein silico\u003c/em\u003e exploration and could also explain why the engineered strains\u0026rsquo; growth rate is diminished. Another common feature in all simulations is the increase of upper and lower glycolysis reactions flux values leading to a greater pool of phosphoenolpyruvate. Such raise could be explained by the higher demand of energy spent on the ectopic kinase\u0026rsquo;s functioning or a general metabolic pull directed to the TCA cycle, as the oxidase may be enhancing the activity of the electron transport chain in the mitochondria. At the same time, differences in the internal fluxes\u0026rsquo; perturbations caused by the expression of ectopic genes can be found too. For example, the oxidative branch reactions in the pentose phosphate pathway (PPP), one of the main NADPH sources in yeast (34), are upregulated in the AtOX simulations but their flux decreases when POS5 activates as the kinase is already providing NADPH to the network. This observation is further reinforced by transcriptomic data, which shows corresponding changes in gene expression and suggests a clear correlation between catalytic activity and transcriptional regulation. Lastly, the TCA cycle fluxes are perturbed differently. Particularly, the NAD-dependent Isocitrate dehydrogenase flux increases to cope with the extra NADH demand due to alternative oxidase while the opposite occurs in the presence of the ectopic NADH kinase.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental data constrained\u003c/strong\u003e \u003cstrong\u003ein silico\u003c/strong\u003e \u003cstrong\u003eanalysis of cytosolic NADH kinase overexpression impact on internal metabolic fluxes (Learn phase)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eIn silico\u003c/em\u003e simulations of the Rol-producing strain co-expressing the cytosolic \u003cem\u003eS. cerevisiae\u003c/em\u003e NADH kinase (cPOS5) gene revealed trends that closely matched previously reported experimental observations (35). First, our genome-scale based simulation predicted just a negligible increase in ATP regeneration rate when the cPOS5 kinase is expressed. This is coherent with the previous study, where ATP/ADP ratio in the cPOS5 strain remained unaltered, indicating that ATP homeostasis is not substantially perturbed (35). Second, consistent with the remarkable increase in the NADPH/NADP ratio observed in (35), our simulations showed a clear increase in the NADPH regeneration capacity, supporting the notion that cPos5 activity effectively redirects reducing equivalents towards the NADPH pool. Finally, both the computational predictions and cultivation data provided in (35) demonstrated a reduction in biomass yield, which can be attributed to the ATP‑dependent phosphorylation of NADH and the resulting partial dissipation of cellular energy. Together, these concordant results reinforce the mechanistic link between cofactor redistribution caused by cPOS5 kinase and the metabolic adjustments observed \u003cem\u003ein vivo\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental data constrained\u003c/strong\u003e \u003cstrong\u003ein silico\u003c/strong\u003e \u003cstrong\u003eanalysis of alternative oxidase overexpression impact on internal metabolic fluxes (Learn phase)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe \u003cem\u003ein silico\u003c/em\u003e analysis of the HcAtOX-overexpressing strain reveals several metabolic trends that closely parallel observations reported for \u003cem\u003eS. cerevisiae\u003c/em\u003e strains ectopically expressing the same alternative oxidase. First, the presence of AOX consistently increased glucose and oxygen turnover rates, indicating enhanced glucose oxidation, a phenomenon also evident from cultivation data in both previous studies. This metabolic shift is accompanied by a reduction in biomass yield, a recurring outcome associated with AOX overexpression. Our model further predicts an increased flux through isocitrate dehydrogenase, aligning with the elevated enzymatic activity assays reported in (36). Similarly, the enhanced mitochondrial regeneration capacity of NADH and NAD\u003csup\u003e+\u003c/sup\u003e predicted in our simulations mirrors the higher intracellular levels of these cofactors measured experimentally in that study. HcAtOX expression also exerts a pronounced influence on mitochondrial metabolism: Most TCA cycle reactions show increased fluxes while glyoxylate cycle fluxes decreases, matching the transcriptional up-regulation of TCA genes and down-regulation of glyoxylate cycle genes described in (36). Finally, although AOX provides an additional respiratory route by uncoupling NADH oxidation from proton translocation\u0026mdash;thereby dissipating energy and contributing to lower biomass yields\u0026mdash;its impact on ATP metabolism appears context-dependent. Our simulations predict a hindered ATP turnover rate, whereas previous studies reported an increased P/O ratio and reduced ATP requirement for biomass formation in \u003cem\u003eS. cerevisiae\u003c/em\u003e AOX strains (35), suggesting that HcAtOX-driven metabolic rewiring may enhance ATP synthesis efficiency under certain conditions. Ultimately, HcAtOX overexpression seems to mitigate well known overflow-metabolism side-effects in \u003cem\u003eS. cerevisiae\u003c/em\u003e growing on glucose. However, this property does not provide a functional advantage in our case, as high glucose concentrations would repress the AOX promoter driving recombinant protein production. On the contrary, this suggests that HcAtOX ectopic overexpression is more likely to impose a metabolic burden on \u003cem\u003eK. phaffii\u003c/em\u003e under the tested growth conditions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental data constrained\u003c/strong\u003e \u003cstrong\u003ein silico\u003c/strong\u003e \u003cstrong\u003eanalysis of co-overexpression of both ectopic genes impact on internal metabolic fluxes (Learn phase)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe simulation of the double‑mutant strain co-expressing cPOS5 and the alternative oxidase (HcAtOX) genes revealed an unexpectedly complex metabolic behavior, as the two ectopic activities appear to perturb the network in opposing directions. While cPOS5 drives NADH consumption and enhances NADPH formation, AOX provides an additional sink for NADH oxidation, and the simultaneous presence of both functions generates flux patterns that are not straightforward to interpret. Despite this apparent antagonism, the dominant features of the flux distribution can be attributed to a pronounced increase in glucose uptake, which supplies elevated levels of both NADH and ATP through glycolysis. This enhanced glycolytic contribution could explain the observed decrease in the proportion of methanol channeled through the dissimilatory pathway, as the cell becomes less dependent on methanol‑derived NADH for energy generation.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eIn this study, we engineered \u003cem\u003eK. phaffii\u003c/em\u003e\u0026rsquo;s redox metabolism by integrating ectopic genes encoding enzymes that convert NADH into NADPH or NAD⁺, aiming to balance the redox equivalents demands required for recombinant protein synthesis and secretion. We demonstrate that heterologous expression of c\u003cem\u003ePOS5\u003c/em\u003e and HcAtOX, encoding an NADH kinase targeted to the cytosol and an alternative NADH oxidase, respectively, can enhance Rol production under methanolic growth conditions. Chemostat cultivations revealed increases in Rol titers ranging among 35\u0026ndash;50% when c\u003cem\u003ePOS5\u003c/em\u003e was overexpressed, either alone or in combination with HcAtOX.\u003c/p\u003e \u003cp\u003eOur results show that the expression level of ectopic genes is a critical determinant of their metabolic impact, influencing both redox cofactor availability and global flux redistribution. Moreover, combining different candidate genetic modifications can lead to additive or even synergistic effects beneficial for recombinant protein production.\u003c/p\u003e \u003cp\u003eThe \u003cem\u003ein silico\u003c/em\u003e initial exploration indicated that metabolic and production phenotypes depend strongly on the carbon source(s) supplied. These findings suggest that the optimal genetic modification strategy is growth condition-dependent; therefore, for each envisaged or constrained bioprocess strategy/condition, a different engineering approach should be identified, e.g. by following the workflow employed in this study. Additional \u003cem\u003ein silico\u003c/em\u003e simulations using the genome-scale metabolic model informed with the experimental data obtained from cultivations of the engineered strains revealed metabolic perturbations widespread the whole metabolic network. The agreement between predicted flux changes and available transcriptomic data further supports the model\u0026rsquo;s accuracy and highlights its value in the DBTL cycle.\u003c/p\u003e \u003cp\u003eBeyond recombinant protein production, the redox engineering strategy explored here can be readily extended to other biotechnological applications in \u003cem\u003eK. phaffii\u003c/em\u003e, particularly in pathways where redox cofactor unbalance becomes a metabolic bottleneck. Nevertheless, the genetic robustness of the redox-engineered strains should be evaluated in long-term cultivations, as redox/energy status may affect cell\u0026rsquo;s lifespan. Finally, assessing the applicability of the workflow developed in this study across different substrates, cultivation modes, and target products will further validate its potential.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003e \u003cb\u003eConflicts of interest\u003c/b\u003e:\u003c/h2\u003e \u003cp\u003eNo potential conflict of interest is reported by the authors.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding:\u003c/h2\u003e \u003cp\u003eThis study was supported by the projects CTQ2016-74959-R, PID2019-104666GB-I00, and PID2022-136936OB-I00 of the Spanish Ministry of Economy, Industry, and Competitiveness. Sergi Monforte was recipient of the PhD fellowship FPI BES-2014-067935 of the Spanish Ministry of Science and Innovation. Eric Anton is recipient of the PhD fellowship FI (2022FI_B_00639) of the AGAUR (Catalan Government. The research group is supported grants 2021-SGR-00143 from the AGAUR.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eEA performed in silico simulations, constructed and assessed the strain co-overexpressing both the NADH kinase and alternative oxidase encoding genes, and contributed to the manuscript drafting. SM built the strains overexpressing an alternative oxidase encoding gene, performed the cultures of these strains, as well as contributing to the manuscript drafting. AQ built the strains overexpressing a NADH kinase encoding gene and assisted in the screening of these strains. JA contributed to the design of the in silico experiments and interpretation of the in silico and experimental results and reviewed the manuscript. PF participated in the conceptual design of the experiments, interpretation of the experimental results and contributed to the manuscript drafting.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll data supporting the findings of this study are available within the paper and its Supplementary Information. Raw data will be made available on request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003e Glick BR. Metabolic load and heterologous gene expression. Biotechnology Advances. 1995. p. 247\u0026ndash;61. doi:10.1016/0734-9750(95)00004-A PubMed PMID: 14537822.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e Ferrer P, Albiol J. \u003csup\u003e13\u003c/sup\u003eC-based metabolic flux analysis of recombinant \u003cem\u003ePichia pastoris\u003c/em\u003e. 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An\u0026agrave;lisi quantitatiu de l\u0026rsquo;efecte de l\u0026rsquo;expressi\u0026oacute; d\u0026rsquo;una lipasa recombinant sobre el metabolisme central de \u003cem\u003ePichia pastoris\u003c/em\u003e. 2013. Universitat Aut\u0026ograve;noma de Barcelona. https://hdl.handle.net/10803/117310\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e Bertels LK, Murillo LF, Heinisch JJ. The pentose phosphate pathway in yeasts\u0026ndash;more than a poor cousin of glycolysis. Biomolecules. 2021. doi:10.3390/biom11050725.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e Hou J, Lages NF, Oldiges M, Vemuri GN. Metabolic impact of redox cofactor perturbations in Saccharomyces cerevisiae. Metab Eng. 2009 Jul;11(4\u0026ndash;5):253\u0026ndash;61. doi:10.1016/j.ymben.2009.05.001.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e Vemuri GN, Eiteman MA, Mcewen JE, Olsson L, Nielsen J. Increasing NADH oxidation reduces overflow metabolism in \u003cem\u003eSaccharomyces cerevisiae\u003c/em\u003e. Proc Natl Acad Sci U S A. 2007 Feb 13;104(7):2402-7. doi: 10.1073/pnas.0607469104.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"microbial-cell-factories","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"micf","sideBox":"Learn more about [Microbial Cell Factories](http://microbialcellfactories.biomedcentral.com/)","snPcode":"12934","submissionUrl":"https://submission.nature.com/new-submission/12934/3","title":"Microbial Cell Factories","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Komagataella phaffii, NADH kinase, alternative oxidase, Rhizopus oryzae lipase, POS5, AOX, redox cofactor balance, metabolic burden, genome-scale metabolic model","lastPublishedDoi":"10.21203/rs.3.rs-9453242/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9453242/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eRecombinant protein extracellular production is a highly demanding process in terms of energy and redox cofactor regeneration. Yeasts such as \u003cem\u003eKomagataella phaffii\u003c/em\u003e (aka. \u003cem\u003ePichia pastoris\u003c/em\u003e) seem to adapt their metabolism by increasing ATP, NADH or NADPH regeneration to compensate for the metabolic burden exerted by such costly process. Earlier studies have proven that increased NADPH regeneration capacity has a positive effect on recombinant protein production when using glucose as sole carbon source, while counteracting increased NADH/NAD\u003csup\u003e+\u003c/sup\u003e ratios has also been proven beneficial.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eTo evaluate the metabolic impact of increasing NADPH and/or NAD\u003csup\u003e+\u003c/sup\u003e regeneration caused by the introduction of a cytosolic NADH kinase and an alternative NADH oxidase, we initially performed \u003cem\u003ein silico\u003c/em\u003e simulations based on the use of a genome-scale model. Next, we investigated the effect of overexpressing the \u003cem\u003eS. cerevisiae\u003c/em\u003e\u0026rsquo;s \u003cem\u003ePOS5\u003c/em\u003e NADH kinase encoding gene (targeted to the cytosol) and an alternative oxidase (\u003cem\u003eAtOX\u003c/em\u003e) in a \u003cem\u003eK. phaffii\u003c/em\u003e strain producing extracellularly a \u003cem\u003eRhizopus oryzae\u003c/em\u003e lipase (Rol) under control of the alcohol oxidase 1 (\u003cem\u003eAOX1\u003c/em\u003e) promoter (P\u003csub\u003e\u003cem\u003eAOX1\u003c/em\u003e\u003c/sub\u003e), thereby assessing the impact of such engineering strategies on recombinant protein production and cell performance under methanol growth conditions. Small-scale cultures allowed us to conclude that such genetic modifications increase protein production significantly under methanol growth conditions. The redox-engineered strains were further characterized in bioreactor chemostat cultures using a glucose/methanol mix (60/40%, w/w) as a carbon source to corroborate their performance. Thereby, up to a 36% increase in Rol specific productivity was observed when overexpressing two copies of \u003cem\u003ePOS5\u003c/em\u003e, whereas AtOX overexpression did not improve production. Notably, combined two-copy \u003cem\u003ePOS5\u003c/em\u003e and AtOX overexpression resulted in a 53% increase in lipase production compared to the reference strain, suggesting that perturbation of the NADH/NAD\u003csup\u003e+\u003c/sup\u003e ratio leads to widespread changes in metabolism ultimately affecting cell growth and product formation, resulting in the observed additive effect. An \u003cem\u003ein-silico\u003c/em\u003e interpretation of the physiological macroscopic data from chemostat cultures using a genome-scale metabolic model allowed us to gain further insights on the metabolic impact of these modifications.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eOur study provides a DBTL cycle workflow generating valuable insights for a quantitative understanding and prediction of cellular response to redox- based perturbations and subsequent test bedding of metabolic engineering applications in \u003cem\u003eK. phaffii\u003c/em\u003e.\u003c/p\u003e","manuscriptTitle":"Metabolic impact of redox perturbations on recombinant protein production in Komagataella phaffii under methanolic growth conditions","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-05-06 09:14:43","doi":"10.21203/rs.3.rs-9453242/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"18613497749706296660027320383385518849","date":"2026-05-05T06:46:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"211103829903931296992922034956929268306","date":"2026-05-04T06:48:11+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"205017377504490529069311296869527235092","date":"2026-04-28T13:31:03+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-28T09:00:32+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-04-24T19:49:05+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-04-24T09:57:00+00:00","index":"","fulltext":""},{"type":"submitted","content":"Microbial Cell Factories","date":"2026-04-18T00:39:26+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"microbial-cell-factories","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"micf","sideBox":"Learn more about [Microbial Cell Factories](http://microbialcellfactories.biomedcentral.com/)","snPcode":"12934","submissionUrl":"https://submission.nature.com/new-submission/12934/3","title":"Microbial Cell Factories","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a414474f-ddaf-4c88-b439-340df0ea483c","owner":[],"postedDate":"May 6th, 2026","published":true,"recentEditorialEvents":[{"type":"reviewerAgreed","content":"18613497749706296660027320383385518849","date":"2026-05-05T06:46:54+00:00","index":13,"fulltext":""},{"type":"reviewerAgreed","content":"211103829903931296992922034956929268306","date":"2026-05-04T06:48:11+00:00","index":12,"fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-05-06T09:14:43+00:00","versionOfRecord":[],"versionCreatedAt":"2026-05-06 09:14:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9453242","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9453242","identity":"rs-9453242","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}