Biochemical Engineering Journal最新文献

{"title":"Model-based optimization of stripping onset in Saccharomyces cerevisiae very-high-gravity ethanol fermentations","authors":"I.I.K. Veloso ,&nbsp;V.T. Mazziero ,&nbsp;D.A. Lemos ,&nbsp;A.J.G. Cruz ,&nbsp;M.O. Cerri ,&nbsp;A.C. Badino","doi":"10.1016/j.bej.2025.110057","DOIUrl":"10.1016/j.bej.2025.110057","url":null,"abstract":"<div><div>Very-high-gravity (VHG) fermentation increases ethanol yield and decreases water use, but it imposes severe osmotic and ethanolic stress on <em>Saccharomyces cerevisiae</em>, which frequently limits overall productivity. In situ ethanol removal through CO₂ gas stripping can mitigate this inhibition. However, the optimal timing for stripping onset under different temperature conditions remains not fully characterized. This study examined the effect of stripping initiation timing in VHG fed-batch fermentations at 28, 30, 32, and 34°C. A mechanistic fermentation model based on Monod-type kinetics with ethanol inhibition terms and coupled gas-liquid mass transfer was estimated using differential evolution. Distinct, temperature-specific ethanol concentration thresholds for stripping onset were identified, which maximized ethanol productivity (<span><math><msub><mrow><mi>P</mi></mrow><mrow><mi>CE</mi></mrow></msub></math></span>). Plateau analysis, based on identification of concentration regions where delayed initiation produced negligible changes in <span><math><msub><mrow><mi>P</mi></mrow><mrow><mi>CE</mi></mrow></msub></math></span>, refined these thresholds to within ±0.5 % of the maximum productivity. This resulted in CO₂ gas savings equivalent to 1.05 ± 0.06 h per fermentation cycle. Model-based time-varying temperature control optimization predicted an increase in ethanol productivity to 12.32 g L⁻¹ h⁻¹. The findings provide a simulation and parameter estimation framework for temperature-integrated extractive control strategies to improve ethanol production in high-biomass VHG fermentations.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110057"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145880162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DLF-BBP: A novel deep learning framework for blood-brain barrier penetrating peptides design","authors":"Lei Huang ,&nbsp;Zhanglong Li ,&nbsp;Zhengtai Li ,&nbsp;Hongjiang Li ,&nbsp;Tiejun Bing ,&nbsp;Yingji Li ,&nbsp;Changyuan Yu","doi":"10.1016/j.bej.2025.110014","DOIUrl":"10.1016/j.bej.2025.110014","url":null,"abstract":"<div><div>The blood-brain barrier (BBB) maintains brain homeostasis by selectively regulating substance exchange but also limits central nervous system (CNS) drug delivery. Blood-brain barrier penetrating peptides (BBPs) are short peptides capable of traversing the BBB, serving as drug carriers or therapeutic agents. Therefore, designing novel BBPs is crucial for overcoming the BBB's limitations and advancing CNS-targeted drug development. In this study, we propose a novel deep learning framework for BBPs design (DLF-BBP). Beginning with a Self-Attention Generative Adversarial Network (SAGAN) model to generate novel candidate BBPs that were evaluated in silico, we subsequently constructed a predictive model based on a Convolutional Neural Network (CNN) for the initial screening of GAN-designed BBPs, achieving an area under the curve (AUC) of 0.988 and an accuracy of 0.954. Further multi-step screening was conducted using physicochemical property analysis, molecular docking, and online platforms. Finally, the filtered candidate GAN-designed BBPs were validated through in vitro BBB penetration assay, leading to the identification of three promising GAN-designed BBPs with high permeation capability, demonstrating penetration comparable to that of the positive control Lixisenatide. Our research offers new possibilities for treating neurological disorders.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110014"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145617517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biodegradation characteristics, pathway elucidation, and toxicity assessment of oxytetracycline-polyethylene combined pollution by two Aspergillus fungi","authors":"Yuan Zhang,&nbsp;Shuicao Liu,&nbsp;Tingting Ding,&nbsp;Wanwan Hao,&nbsp;Shuwen Ren,&nbsp;Jingwen Min,&nbsp;Congyang Zou","doi":"10.1016/j.bej.2025.110027","DOIUrl":"10.1016/j.bej.2025.110027","url":null,"abstract":"<div><div>The widespread use of antibiotics and agricultural microplastics has become a global environmental issue, yet studies on the degradation of their combined pollution remain limited. Here, two functional strains, <em>Aspergillus</em> sp. DL-1 and DL-2, selected from 10 isolates and identified via ITS sequencing and phylogenetic analysis, were investigated for their ability to degrade oxytetracycline (OTC)–polyethylene (PE) combined pollution under optimal conditions. The OTC-PE combined treatment with fungi exhibited superior degradation performance. The OTC degradation rate in the OPD2 treatment reached 96.33 ± 0.36 %, while the weight loss rates of PE in the OPD1 and OPD2 were 10.83 ± 0.53 % and 8.92 ± 0.51 %, respectively. Furthermore, the WCA of the PE films in the combined treatments remained consistently lower than in the single-PE treatments, accompanied by significant changes in surface morphology and structure. Under the treatment of the strains, OTC can be degraded through multiple pathways, including decarbonylation, decarboxylation, deamination, demethylation, and dehydration. Specifically, biodegradation by DL-1 and DL-2 markedly lowered the developmental toxicity, mutagenicity, and bioaccumulation potential of OTC and its intermediates, thereby alleviating the environmental toxicity burden. In parallel, PE cleaved the chain under the action of microbial enzymes to generate low molecular hydrocarbons, which are further oxidized to carboxylic acids and other intermediate products. These findings provide a novel approach for addressing compound pollution in agricultural environments.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110027"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145682448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Addressing temperature gradient challenge in scaling up solid-state fermentation: A strategy using thermophilic strains for biosurfactant production","authors":"Jose Bueno-Mancebo,&nbsp;Adriana Artola,&nbsp;Syeda Amna Farooq,&nbsp;Raquel Barrena,&nbsp;Teresa Gea","doi":"10.1016/j.bej.2025.110037","DOIUrl":"10.1016/j.bej.2025.110037","url":null,"abstract":"<div><div>Solid-state fermentation (SSF) is a promising approach for sustainable bioproduction production, particularly when using organic waste as a substrate. However, temperature gradients inherent to large-scale SSF often reduce process efficiency. This study developed a strategy for designing of a lipopeptide biosurfactant production process<em>,</em> scaling SSF in packed-bed bioreactors by thermophilic strains using winterisation oil cake (WOC) and sugarcane molasses (MOL) as nutrient sources. Fermentations at lab-scale (0.5 L) and pilot-scale (50 L) demonstrated a robust and reproducible process despite temperature changes from microbial activity. Among the strains tested, <em>Bacillus subtilis</em> CBI-7S1 was the highest producer, with optimal substrates amounts of 24 g of WOC and 12 g of MOL, producing 24.9 mg of crude lipopeptides per gram of dry matter (DM) at 0.5 L scale. These conditions were reproduced at pilot scale, with concentrations from 12 to 24 mg g⁻¹ DM. Moisture content strongly influenced biosurfactant production, while oxygen consumption was a reliable monitoring parameter. Surface tension was evaluated under different pH, temperature, and salinity, and mass spectrometry identified surfactin, iturin, and fengycin congeners. These findings provide insights into overcoming temperature gradients during scale-up and show that thermophilic strains enable biosurfactant production under SSF at pilot scale. This approach enhances SSF technological maturity, supporting its broader use in sustainable production of targeted metabolites across diverse bioprocesses.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110037"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145734006","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the effect of media composition on growth and mAb production in CHO cells using a piecewise hybrid dFBA-PLS framework","authors":"Zahra Negahban ,&nbsp;Ali Ghodba ,&nbsp;Anne Richelle ,&nbsp;Chris McCready ,&nbsp;Valerie Ward ,&nbsp;Hector Budman","doi":"10.1016/j.bej.2025.110013","DOIUrl":"10.1016/j.bej.2025.110013","url":null,"abstract":"<div><div>In this study, we present a hybrid modeling framework that integrates piecewise Partial Least Squares (PLS) regression with Dynamic Flux Balance Analysis (dFBA) to simulate and optimize Chinese Hamster Ovary (CHO) cell fed-batch culture. Twenty-four Ambr15 experiments were conducted to systematically vary feed and inoculum compositions. Time-resolved metabolite, biomass, and Monoclonal antibodies (mAb) concentrations were collected and modeled. The hybrid model achieved high prediction accuracy (Normalized Mean Squared Error (NMSE) <span><math><mo>&lt;</mo></math></span> 0.15 for most metabolites) and provided interpretable flux profiles. Multivariate analysis revealed consistent metabolic signatures tied to media formulation, where specific feed–inoculum combinations drove shifts in glycolysis, TCA cycle flux, and nitrogen metabolism. These insights demonstrate the model’s capacity to capture key metabolic adaptations and support data-driven media optimization in CHO cell culture.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110013"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145594588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An adsorption-crosslinking immobilization approach for enhancing the operational stability of D-allulose 3-epimerase immobilized on chitosan-modified resin","authors":"Yingying Zheng ,&nbsp;Hongwei Liu ,&nbsp;Shumin Guo ,&nbsp;Jingwen Fei ,&nbsp;Junqing Wang ,&nbsp;Ruiming Wang ,&nbsp;Piwu Li","doi":"10.1016/j.bej.2025.110039","DOIUrl":"10.1016/j.bej.2025.110039","url":null,"abstract":"<div><div><span>D</span>-allulose is a naturally occurring rare ketohexose that serves as a potential sucrose substitute. In this study, we developed a novel adsorption-crosslinking immobilization strategy based on a chitosan (CTS)-modified resin. After screening 15 resin types, D213 and ZGA351 were selected as carriers, and the CTS-modified carriers were crosslinked with glutaraldehyde to immobilize <span>D</span>-allulose 3-epimerase (DAE; EC 5.1.3.30). CTS modification significantly enhanced the performance of the immobilized enzymes, with the activity recovery rates of D213@CTS@DAE and ZGA351@CTS@DAE increasing compared to the unmodified carriers. The thermal, pH, and storage stabilities of the immobilized enzymes were significantly higher than those of free enzymes. D213@CTS@DAE retained high initial activity across multiple recycling and reuse cycles and sustained productivity in a packed-bed reactor after 29 days of continuous operation. The carrier material could be efficiently regenerated and reused. After regeneration, the immobilized enzyme maintained a high activity retention rate after five cycles. The findings provide a green biocatalytic solution for <span>D</span>-allulose production, which enhances the practicality and commercial viability of the immobilized enzyme.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110039"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hybrid section-based modeling of gas-phase hydrodynamics in biotrickling filters: Influence of packing material on residence time distributions","authors":"Felipe A. Carreño-López ,&nbsp;Patricio A. Moreno-Casas ,&nbsp;Felipe Scott ,&nbsp;Alberto Vergara-Fernández ,&nbsp;José Sierra-Pallares ,&nbsp;Raúl Muñoz","doi":"10.1016/j.bej.2025.110049","DOIUrl":"10.1016/j.bej.2025.110049","url":null,"abstract":"<div><div>In this study, the gas-phase hydrodynamics within a 6.5 L biotrickling filter were quantified using pulse-injection residence time distributions (RTDs) recorded with a cost-effective metal oxide (MOx) sensor. This was conducted at a constant gas flow rate of 8 L min⁻¹ across trickling liquid velocities (TLV) ranging from 0 to 10 <span><math><msup><mrow><mtext>m h</mtext></mrow><mrow><mo>−</mo><mn>1</mn></mrow></msup></math></span>, using three different packings: polyurethane foam, Tri-Packs Jaeger spheres, and wood bark. A hybrid section-based model was developed and evaluated against two closed-closed axial dispersion variants (AD<span><math><msub><mrow></mrow><mrow><mi>c</mi><mi>c</mi></mrow></msub></math></span>+CSTR and AD<span><math><msub><mrow></mrow><mrow><mi>c</mi><mi>c</mi></mrow></msub></math></span>+PFR). Model discrimination using AIC/BIC and <span><math><msup><mrow><mi>R</mi></mrow><mrow><mn>2</mn></mrow></msup></math></span> indicated that the optimal model description is contingent upon the packing material and operating regime: the hybrid model most accurately represented foam and most wood cases, whereas AD<span><math><msub><mrow></mrow><mrow><mi>c</mi><mi>c</mi></mrow></msub></math></span>+CSTR was sufficient for spheres under irrigation, and AD<span><math><msub><mrow></mrow><mrow><mi>c</mi><mi>c</mi></mrow></msub></math></span>+PFR consistently underperformed. Employing the optimal model in each scenario, the Peclet number increased with TLV for PUF and spheres (with diminishing returns at the highest TLV), whereas wood exhibited a low Peclet number and non-monotonic behavior. By explicitly partitioning the mean residence time between the buffer zones and the bed, the hybrid model effectively separates the boundary hold-up from the core dispersion, yielding a more precise representation of the packed bed Peclet number. Overall, MOx-based RTD measurements, in conjunction with model comparison, offer a cost-effective in situ diagnostic tool for differentiating boundary mixing from bed transport, thereby informing packing selection and operating parameters in gas-phase biofiltration.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110049"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioconversion kinetics and valorisation of oil palm empty fruit bunches for biomethane production and lignin derived compounds","authors":"Irnia Nurika ,&nbsp;Eka Nur Shabrina ,&nbsp;Nurul Azizah ,&nbsp;Sri Suhartini ,&nbsp;Guy C. Barker ,&nbsp;Timothy D.H. Bugg","doi":"10.1016/j.bej.2025.110053","DOIUrl":"10.1016/j.bej.2025.110053","url":null,"abstract":"<div><div>This study assesses the facultative anaerobe <em>Comamonas testosteroni</em> as a mild, chemical-free pre-treatment for valorising oil palm empty fruit bunches (OPEFB) into biomethane and lignin-derived aromatics. Incubation with 2 % (v/v) <em>C. testosteroni</em> for 7 days at 30 °C, reduced lignin to 15.67 % (33.42 % removal), while retaining high cellulose (53.48–56.19 %) and hemicellulose (up to 16.21 %). Pre-treated OPEFB showed 20 % weight loss (vs 8 % in controls) and a 51 % rise in total soluble phenols, evidencing active lignin depolymerisation. GC–MS of liquor and solids qualitatively confirmed lignin breakdown and identified representative low-molecular-weight products (e.g., phenol, betulin, acetic acid and benzoxazole), supporting co-product potential. In biochemical methane potential tests, the pre-treated residue achieved a specific methane potential of 0.173 m³ CH₄ kg⁻¹ VS, an 85 % increase over the abiotic residue and higher than non-treated OPEFB. Kinetic fitting with Transference, Modified Gompertz and Logistic models yielded good agreement (R² up to 0.92), with Modified Gompertz best capturing cumulative methane production and the shortest lag phases. A simple energy check indicates a favourable margin: the incremental methane (ΔSMP ≈ 0.08 m³ CH₄ kg⁻¹ VS) equates ∼0.8 kWh kg⁻¹ VS, whereas estimated mixing for pre-treatment is ∼0.017–0.034 kWh kg⁻¹ VS. Overall <em>C. testosteroni</em> pre-treatment enables dual valorisation (biomethane plus aromatics) under mesophilic conditions, offering a practical route for integrated OPEFB biorefineries.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110053"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145836647","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insights into microbial and sorptive regulation of chlorpyrifos-bispyribac dissipation in floating treatment wetlands","authors":"Kashaf Farooq ,&nbsp;Muhammad Arslan ,&nbsp;Samina Anwar ,&nbsp;Maryam Noor ,&nbsp;Muhammad Afzal","doi":"10.1016/j.bej.2025.110045","DOIUrl":"10.1016/j.bej.2025.110045","url":null,"abstract":"<div><div>Agricultural runoff remains one of the most persistent threats to water quality worldwide. Floating wetlands (FWs), when designed appropriately, offer a promising nature-based solution. Here, we show how biologically and sorptively active FWs can remove two agrochemicals—chlorpyrifos (CPF) and bispyribac‑sodium (BIS), under response surface methodology (RSM)‑backed optimal operation (pH 8; 35 °C; 10 mg L⁻¹ each; 1 % glucose). FWs built with <em>Phragmites australis</em> (common reed) were amended with a defined consortium (CB2H, 1 % v/v), plant‑derived biochar (1.5 % w/v), biochar‑immobilized CB2H (1.5 % w/v), and nutrients (N 25 mg L⁻¹, P 25 mg L⁻¹, K 20 mg L⁻¹). CPF and BIS declined exponentially, fitting pseudo‑first‑order kinetics with adsorption component (high S, K_d in the immobilized system). CPF disappeared fastest in the consortium‑only and biochar‑immobilized treatments (k = 0.07 and 0.09 day⁻¹), resulting in &gt; 99 % removal (&gt;9.9 mg L⁻¹) by day 20; BIS peaked at &gt; 84 % (8.4 mg L⁻¹) with immobilized cells. FTIR shifts (∼2920–2840, 2800 cm⁻¹) and new C<img>O bands (1600–1800 cm⁻¹) indicated hydrogen bonding in the <em>Phragmites</em> biochar. Approximately 40 % of the CPF loss in the controls was abiotic (sorption/photolysis/hydrolysis). Chemical oxygen demand and ancillary pollutants also declined. Enhanced performance was supported by microbial colonization within biochar pores. The study provides key design constants (k, K_d), positioning engineered FWs as a scalable nature-based technology for pesticide-laden agricultural runoff.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110045"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improvement of furfural tolerance and butanol production in Clostridium beijerinckii ART44 through overexpression of Class I heat shock proteins","authors":"Chaoyue Shi ,&nbsp;Caolin Liu ,&nbsp;Xin Cheng,&nbsp;Yujie Jing,&nbsp;Qinghua Zhang,&nbsp;Wenjie Lai,&nbsp;Hanguang Li","doi":"10.1016/j.bej.2025.110047","DOIUrl":"10.1016/j.bej.2025.110047","url":null,"abstract":"<div><div>The conversion of lignocellulosic hydrolysate into butanol offers a significant approach to reduce environmental pollution and the greenhouse effect. However, the presence of inhibitors in the hydrolysate restricts its utilization efficiency. In this study, we investigated the role of Class I heat shock proteins (HSPs) in <em>Clostridium beijerinckii</em> ART44 under furfural stress. Notably, the <em>grpE</em> gene was identified as being significantly upregulated under this condition. Therefore, to enhance the furfural tolerance and butanol production, the <em>grpE</em> gene was overexpressed. With this effort, the resulting recombinant strain, named as ART44/<em>grpE</em>, exhibited a 73.75 % increase in butanol production (5.34 ± 0.29 g/L) and a 53.99 % increase in total solvent production (7.67 ± 0.16 g/L) under 3.0 g/L furfural stress, compared to the wild-type strain ART44. Furthermore, mechanistic studies revealed that <em>grpE</em> overexpression reduced cell surface hydrophobicity, minimized the leakage of intracellular macromolecules, and increased the proportion of saturated fatty acids in the cell membrane. These alterations contributed to enhancing cell viability, improving butanol biosynthesis enzyme activity, and reducing membrane permeability under furfural stress. Collectively, the results demonstrate that overexpressing <em>grpE</em> in <em>C. beijerinckii</em> ART44 is an effective and attainable approach to obtain a high furfural tolerance strain, and then improving its butanol production from lignocellulose hydrolysate.</div></div>","PeriodicalId":8766,"journal":{"name":"Biochemical Engineering Journal","volume":"227 ","pages":"Article 110047"},"PeriodicalIF":3.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145786986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}