Biotechnology Letters最新文献

{"title":"Optimization and comparative evaluation of fungal laccase-based biobleaching of pineapple leaf pulp: toward sustainable non-wood pulp processing using Paramarasmius palmivorus.","authors":"Sri Harjati Suhardi, Dea Ashari, Hendro Risdianto, Mamat Kandar, Isty Adhitya Purwasena, Dani Permana","doi":"10.1007/s10529-026-03769-7","DOIUrl":"10.1007/s10529-026-03769-7","url":null,"abstract":"<p><p>The conventional chemical bleaching of pulp generates substantial environmental burdens due to toxic effluents and high chemical consumption. This study explores a sustainable alternative through the enzymatic biobleaching of pineapple leaf fiber (PALF) pulp using crude laccase extracted from Paramarasmius palmivorus, a white-rot fungus. The effects of enzyme dosage (1-10 U/g pulp) and reaction time (2-10 h) on pulp quality were systematically optimized using Response Surface Methodology (RSM). Key pulp properties, including Kappa number, brightness, and intrinsic viscosity, were evaluated and compared to chemical bleaching using hydrogen peroxide (H₂O₂). Results showed that laccase treatment reduced the Kappa number by up to 57% (to 3.91) and enhanced brightness by 29.5% ISO, achieving comparable delignification to H₂O₂ treatment (Kappa = 4.07). Although the effect of dosage and time on brightness and viscosity was not statistically significant (p > 0.05), enzyme dosage had a pronounced influence on lignin removal (p < 0.05). Enzymatic bleaching preserved cellulose integrity with only 22.5% viscosity loss, markedly lower than the 34% loss observed under chemical bleaching. Optimization predicted optimal conditions at 9.8 U/g pulp and 2.1 h, balancing high delignification and minimal cellulose degradation. These findings demonstrate the potential of P. palmivorus laccase as an eco-friendly and efficient biocatalyst for partial or total replacement of chemicals in non-wood pulp bleaching, supporting the transition toward greener pulp industries.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148697040","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production, biochemical characterization, and kinetic/thermodynamic analysis of a novel keratinase from Bacillus sp. MSGU2024.","authors":"Fatemeh Salimi, Eisa Jorjani, Mahmoud Salehi, Matia Sadat Borhani, Hossein Sabouri","doi":"10.1007/s10529-026-03761-1","DOIUrl":"10.1007/s10529-026-03761-1","url":null,"abstract":"<p><p>Due to their distinct characteristics compared to other proteases, Keratinases present a promising alternative for various industrial applications. This study focused on isolating and screening keratinolytic bacteria from poultry waste-contaminated soil, optimizing keratinase production and thoroughly analyzing the biochemical, physicochemical, and thermodynamic properties of the keratinase produced by the isolated strain, Bacillus sp. MSGU2024. Optimized culture conditions resulted in a 2.9-fold increase in keratinase production compared to the initial unoptimized basal medium. The enzyme exhibited optimal activity at 55 <math><mmultiscripts><mrow></mrow> <mrow></mrow> <mo>∘</mo></mmultiscripts> </math> C and pH 8. The keratinase displayed stability in the presence of reducing agents, surfactants, and organic solvents, with stability enhanced by 1.5-2.5 times in the presence of non-ionic detergents such as Tween 20 and Tween 80. Its phenylmethylsulfonyl fluoride (PMSF) inhibition confirmed its classification as a serine protease. The enzyme's <math><msub><mi>K</mi> <mi>m</mi></msub> </math> and <math><msub><mi>V</mi> <mtext>max</mtext></msub> </math> values, 0.102 mM and 0.09 <math><mi>μ</mi></math> M·min^-1 respectively, indicated high substrate affinity and catalytic efficiency. Furthermore, the enzyme's half-life under varying temperature and pH conditions underscored its robustness. The calculated Z-value revealed that the D-value decreased tenfold with a 5.62 <math><mmultiscripts><mrow></mrow> <mrow></mrow> <mo>∘</mo></mmultiscripts> </math> C rise in temperature. The thermodynamic analysis provided key insights, with the activation energy for denaturation ( <math><msub><mi>E</mi> <mtext>d</mtext></msub> </math> ) measured at 254.6 kJ <math><mo>·</mo></math> mol <math><mmultiscripts><mrow></mrow> <mrow></mrow> <mrow><mo>-</mo> <mn>1</mn></mrow> </mmultiscripts> </math> . Gibbs free energy ( <math><mrow><mi>Δ</mi> <msup><mi>G</mi> <mo>∗</mo></msup> </mrow> </math> ), entropy ( <math><mrow><mi>Δ</mi> <msup><mi>S</mi> <mo>∗</mo></msup> </mrow> </math> ), and enthalpy ( <math><mrow><mi>Δ</mi> <msup><mi>H</mi> <mo>∗</mo></msup> </mrow> </math> ) values ranged from 101.59 to 108.57 kJ <math><mo>·</mo></math> mol <math><mmultiscripts><mrow></mrow> <mrow></mrow> <mrow><mo>-</mo> <mn>1</mn></mrow> </mmultiscripts> </math> , 436.02 to 443.65 J <math><mo>·</mo></math> mol <math><mmultiscripts><mrow></mrow> <mrow></mrow> <mrow><mo>-</mo> <mn>1</mn></mrow> </mmultiscripts> </math> <math><mo>·</mo></math> K <math><mmultiscripts><mrow></mrow> <mrow></mrow> <mrow><mo>-</mo> <mn>1</mn></mrow> </mmultiscripts> </math> , and 249.32 to 249.41 kJ <math><mo>·</mo></math> mol <math><mmultiscripts><mrow></mrow> <mrow></mrow> <mrow><mo>-</mo> <mn>1</mn></mrow> </mmultiscripts> </math> , respectively. These findings highlight the enzyme's stability and efficiency, positioning it as a strong candidate for diverse biotechnological applications.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148677029","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Potential health aspect of fermented Spirulina product with Lacticaseibacillus rhamnosus.","authors":"Akif Emre Kavak, Didem Balkanlı, Osman Sağdıç","doi":"10.1007/s10529-026-03770-0","DOIUrl":"10.1007/s10529-026-03770-0","url":null,"abstract":"<p><p>Bio-based fermented products have recently become increasingly important for both human health and sustainability due to the growing interest in healthy sustainable diets. In this study, spirulina was used as a substrate for the cultivation of Lacticaseibacillus rhamnosus 23.2 in a 3 L bioreactor and emphasized the evaluation of the potential health effects of the obtained bioactive products. The antioxidant potential of bioactive products has been extensively studied using assays like DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radicals, demonstrating its capacity to scavenge free radicals and mitigate oxidative stress. MTT and cell migration (scratch) assays reduced viability of Caco-2 cells and hindered cell migration cancer cells FS compared to unfermented spirulina (unFS) revealed that FS significantly reduced cell viability and migration in a dose-dependent manner, with the strongest effects observed at 5X dilution. In contrast, unFS showed weaker bioactivity. These findings highlight fermentation as a promising biotechnological approach to enhance the functional properties of natural compounds for cancer prevention and treatment. To summarize, the current study revealed that the potential of spirulina biomass to be a suitable media substrate for L. rhamnosus 23.2 strain. These findings will provide awareness toward milestone the health and food sector industrialization of FS products.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148668239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Steady expression of heterologous genes in insect cells based on rebuilt Bacmid vectors.","authors":"Wenting Song, Kaiwei Shen, Xiaojie Ma, Yingzhe Yu, Qin Chen, Jian Chen","doi":"10.1007/s10529-026-03768-8","DOIUrl":"10.1007/s10529-026-03768-8","url":null,"abstract":"<p><p>The genetic instability of genes transposed into the baculovirus genome by Bacmid technology, often caused by mini-F replicon, raises a crucial challenge for large scale production of recombinant proteins via serial passages in insect cells. To address this problem, we reconstructed Bacmid vector by decoupling the mini-F replicon from Tn7 transposition site. Two Bacmid variants, BacDC1 and BacDC2, were generated through homologous recombination. In BacDC1, the KanR-lacZα-attTn7 cassette remains in its original position while the mini-F replicon is relocated to the chitinase/v-cathepsin loci. Conversely, BacDC2 undergoes the opposite rearrangement. Using the classical Bacmid technology, two genes encoding enhanced green fluorescent protein (eGFP) and nano-luciferase (Nluc) C-terminally tagged spike 1 (S1) protein of porcine epidemic diarrhea virus (PEDV) were cloned as reporters to assess stability. Fluorescence observation analysis indicated that the genetic stability of cloned genes was markedly improved by the Bacmid variant-derived baculoviruses with sustained expression for up to 20 passages, appropriately 2-3 times longer than the wild-type control. Similarly, western blot and luciferase activity analysis demonstrated the S1-Nluc secretion from Sf9 cells infected with the dual expression variants remained stable over 10 passages, whereas the control showed significant expression decline by passage 5. These data suggest that decoupling the mini-F replicon from Tn7 transposition site significantly stabilizes cloned gene expression. The Bacmid variants thus represent promising vectors for industrial recombinant protein production and gene therapy applications.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148652822","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Carotenoid production by Metschnikowia sp.: isolation from environmental samples and optimization of culture conditions.","authors":"M Mokhtari, M Mohammadi-Sichani, M Madani","doi":"10.1007/s10529-026-03767-9","DOIUrl":"10.1007/s10529-026-03767-9","url":null,"abstract":"<p><p>Carotenoids are valuable natural pigments with antioxidant and health-promoting properties. Growing concerns regarding the toxicity and environmental persistence of synthetic colorants have increased interest in safe and sustainable natural alternatives. Compared with plant-derived carotenoids, microbial production offers several practical advantages, including rapid growth, year-round cultivation, utilization of agro-industrial substrates, and simplified downstream processing. This study aimed to isolate carotenoid-producing microorganisms from environmental samples, identify the most productive isolate, and optimize culture conditions for enhanced carotenoid biosynthesis, with particular emphasis on Metschnikowia sp. Five pigmented isolates were recovered, among which Metschnikowia sp. (100% identity, 90% query coverage; GenBank accession no. MZ622714) exhibited the highest carotenoid production (A450 = 4.2) and was selected for further investigation. Using the Taguchi experimental design, the highest biomass yield (10.67 ± 0.58 mg/mL) was obtained in a medium containing glucose and yeast extract at pH 6 and 20 °C, whereas maximum carotenoid production (6.48 µg/g) was achieved under Molasses and peptone (or the correct optimized medium), at pH 7 and 20 °C. Notably, carotenoid biosynthesis was enhanced under stress-associated conditions rather than those favoring maximal biomass formation. UV-Vis spectroscopy and HPLC analyses confirmed carotenoid production, with a dominant chromatographic peak (retention time ⁓21.4 min) consistent with an astaxanthin-like compound. These findings demonstrate the potential of Metschnikowia sp. as a promising non-conventional yeast for sustainable production of natural carotenoids and provide a foundation for future development of industrial microbial bioprocesses.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148598601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic engineering of tobacco plants using Carica papaya NAC transcription factors: a biotechnology tool to improve plant metabolite production.","authors":"Alberto G Aguilar-Santana, Alejandro Pereira-Santana, Francisco Guillén-Chable, Miguel Herrera-Alamillo, Fabiola Escalante-Erosa, Enrique Castano, Rocío Borges-Argaez, Luis Carlos Rodríguez-Zapata","doi":"10.1007/s10529-026-03757-x","DOIUrl":"10.1007/s10529-026-03757-x","url":null,"abstract":"<p><p>Plant secondary metabolism includes the biosynthesis of a wide array of molecules with significant biological and biotechnological applications. However, the regulatory mechanisms governing secondary metabolite production remain largely elusive. Key master regulators of gene expression, such as transcription factors (TFs), are expected to play a central role in controlling the biosynthesis of these compounds. In this context, the present study addresses a critical knowledge gap by elucidating the regulatory role of Carica papaya NAC transcription factors (CpNACs) in reprogramming secondary metabolites in Nicotiana tabacum. Thus, this work advances our understanding of plant metabolic engineering by identifying molecular links between transcription factor regulation and metabolite accumulation. Methanolic extracts from transformed and non-transformed plants were analyzed by gas chromatography-mass spectrometry (GC-MS). The results showed altered accumulation of some specialized metabolites. Specifically, targeted metabolomics revealed increased accumulation of phytosterols, including campesterol, β-sitosterol, and stigmasterol. Non-targeted metabolomics further indicated extensive metabolic reprogramming, affecting pools of organic acids, steroids, and lipids. Furthermore, transformed seedlings showed elevated levels of phenylpropanoids (e.g., coumarin, sinapaldehyde), alkaloids (e.g., tropinone, allocryptopine), and derivatives of aromatic amino acids (e.g., tryptamine, phenylacetic acid). The increased abundance of key metabolic intermediates, such as acetate and acetyl-CoA, suggests a redirection of metabolic flux toward the biosynthesis of these specialized compounds. Our findings suggest that CpNAC transcription factors may regulate specialized metabolite biosynthetic pathways, potentially by modulating metabolic flux at specific enzymatic steps within these networks. This pioneering study proposes an alternative that could improve the performance and productivity of biotechnological processes for obtaining valuable compounds from plants, highlighting the potential of transcription factor-based approaches to manipulate complex plant metabolic networks.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148590098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biosorption of rare earth elements by Escherichia coli displaying a cyclizable terbium(III)-binding peptide.","authors":"Fumika Kojima, Masato Hoshino, Miho Yoshimura, Kaneyoshi Yamamoto","doi":"10.1007/s10529-026-03766-w","DOIUrl":"10.1007/s10529-026-03766-w","url":null,"abstract":"<p><p>Efficient recovery of rare earth elements (REEs) is essential for sustainable technologies. We evaluated a synthetic cyclizable peptide (peptide 2) for bacterial biosorption. Fluorescence resonance energy transfer analysis, utilizing non-linear regression models, revealed that peptide 2 possesses high affinity for middle-range lanthanides, such as Tb³⁺, Gd³⁺, and Dy³⁺. Disulfide-mediated cyclization significantly enhanced binding affinity, yielding a dissociation constant of 2.6 nM for Tb³⁺. To develop a practical biosorbent, we engineered Escherichia coli to display peptide 2 on its outer membrane via the OmpC protein. These recombinant strains exhibited a standardized adsorption capacity of up to 1.11 mg Tb/g dry biomass, representing a statistically significant improvement of up to 33% compared to the native OmpC control. These results highlight the potential of surface-displayed cyclizable peptides for developing efficient, REE-specific biosorbents.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148560901","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improving expression of Photobacterium sp. JT-ISH-224 α2,6-sialyltransferase using ProteinMPNN and machine learning guided combinatory mutagenesis.","authors":"Qingshi Wen, Huang Li, Weiyi Wang, Ziyi Yan, Pengpeng Yang, Dong Liu, Wei Zhuang, Cheng Zheng, Hanjie Ying","doi":"10.1007/s10529-026-03756-y","DOIUrl":"10.1007/s10529-026-03756-y","url":null,"abstract":"<p><p>Industrial-scale production of 6'-sialyllactose (6'-SL), a sialylated human milk oligosaccharide, has recently attracted increasing attention. As a crucial enzyme for both microbial synthesis approaches and enzymatic catalysis, several α2,6-sialyltransferases (α2,6-SiaTs) have been reported and characterized. However, the low expression levels and poor stability of α2,6-SiaTs remain major bottlenecks in the construction of efficient 6'-SL production systems. In this study, we applied a deep-learning-based protein generative model, ProteinMPNN, to redesign an α2,6-SiaT from Photobacterium sp. JT-ISH-224 (pst6-224), resulting in a marked improvement in its heterologous expression in Escherichia coli BL21(DE3). Furthermore, machine learning-guided combinatorial mutagenesis was employed to restore catalytic activity in the redesigned enzyme. The expression level of the final variant was increased by approximately tenfold compared with that of the wild-type pst6-224, and a higher 6'-SL titer was achieved in a 1 L cascade catalytic system starting from CMP. This work demonstrates the potential of machine learning-assisted protein engineering and provides an α2,6-SiaT variant with high expression levels for the biological synthesis of 6'-SL.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148560491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of mannosylated hrsACE2 by baculovirus expression system; the way to remove and redirect SARS-CoV-2 viral particles to macrophages.","authors":"Hanie-Sadat Emami, Bahareh Vakili, Nafiseh Esmaeil, Marc G Aucoin, Ali Jahanian-Najafabadi","doi":"10.1007/s10529-026-03755-z","DOIUrl":"10.1007/s10529-026-03755-z","url":null,"abstract":"<p><strong>Purpose: </strong>The immune system combats viral infections through innate and adaptive responses, with macrophages and dendritic cells playing key roles in pathogen clearance. Neutralizing antibodies aid viral elimination but have variable efficacy against SARS-CoV-2, which uses ACE2 for entry. Human recombinant soluble ACE2 (hrsACE2) acts as a decoy to block viral entry but has limitations in affinity and immune uptake. We produced mannosylated hrsACE2 (Mann-hrsACE2) via a baculovirus expression system to enhance uptake by innate immune cells, aiming to improve viral neutralization and stimulate cell-mediated immunity against SARS-CoV-2.</p><p><strong>Methods: </strong>Mann-hrsACE2 protein was produced in Sf9 insect cells using recombinant baculovirus, purified by Ni-NTA chromatography, and verified by SDS-PAGE and Western blot. Its interaction with macrophages was assessed by FITC-labeling and analyzed through fluorescence microscopy and flow cytometry.</p><p><strong>Results: </strong>Expressed Mann-hrsACE2 displayed an upward shift SDS-PAGE and Western blot analysis. Flow cytometry revealed macrophage uptake of Mann-hrsACE2 at 3, 28, and 98% corresponding to concentrations of 1, 10, and 51 nM, respectively.</p><p><strong>Conclusion: </strong>The increased molecular weight of the expressed protein confirmed its glycosylation. Its mannosylation enabled efficient uptake by macrophages, key antigen-presenting cells in the immune system. Furthermore, by binding to SARS-CoV-2, the protein not only inhibits viral entry but also redirects the virus to macrophages for enhanced immune clearance.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148519843","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced biodiesel production from Fusarium oxysporum via mutagenesis and process optimization using bagasse hydrolysate as a sustainable carbon source.","authors":"Sayeda Abdelrazek Abdelhamid, Einas Hamed El-Shatoury, Mohsen Selim Asker, Samir Khalef Abd-El-Aal, Ashraf Gamil Attallah, Sahar Saleh Mohamed, Mostafa Mostafa Abo Elsoud","doi":"10.1007/s10529-026-03754-0","DOIUrl":"10.1007/s10529-026-03754-0","url":null,"abstract":"<p><strong>Background: </strong>Biodiesel offers a sustainable alternative to fossil fuels with reduced greenhouse gas emissions; however, its commercial viability is constrained by high production costs. This study aimed to develop a cost-effective strategy for enhanced lipid and biodiesel production using Fusarium oxysporum NRC 2017, cultivated on sugarcane bagasse hydrolysate enzymatically saccharified by Bacillus cereus 3SME, as an inexpensive carbon source.</p><p><strong>Methods: </strong>To improve lipid accumulation, the fungal strain was subjected to chemical and physical mutagenesis using gamma radiation (Ɣ-rays), ethidium bromide (EtBr), ethyl methane sulfonate (EMS), and sodium azide (NaN₃). Genetic variation between the wild type and induced mutants was confirmed by inter-simple sequence repeat (ISSR) analysis. The highest lipid-producing mutant, F. oxysporum NRC 2017-1, derived from gamma radiation, was selected for further optimization. Critical process parameters were optimized using response surface methodology (RSM) to maximize lipid yield.</p><p><strong>Results: </strong>Under optimized conditions, the mutant strain achieved a maximum lipid yield of 3.81 g/L, representing a substantial increase over the wild type. Gas chromatography analysis revealed a favorable fatty acid methyl ester (FAME) profile, dominated by C16-C18 fatty acids, which meets the key compositional requirements for high-quality biodiesel. Furthermore, the physicochemical properties of the produced biodiesel, including cetane number, viscosity, and density, were within the recommended limits of international standards (ASTM D6751 and EN 14214).</p><p><strong>Conclusion: </strong>This study presents an integrated and sustainable approach combining strain improvement via mutagenesis, valorization of lignocellulosic agricultural waste, and statistical process optimization. The findings highlight the potential of F. oxysporum NRC 2017-1 as a promising candidate for scalable, cost-effective biodiesel production.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"48 4","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148496979","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}