Applied Microbiology and Biotechnology最新文献

{"title":"Ammonium and nitrate fluxes drive metabolic functional shifts in tap water biofilms","authors":"Binessi Edouard Ifon,&nbsp;Wei Zhang,&nbsp;Zhongbo Zhou,&nbsp;Okugbe Ebiotubo Ohore,&nbsp;Yeyuan Xiao","doi":"10.1007/s00253-026-13905-y","DOIUrl":"10.1007/s00253-026-13905-y","url":null,"abstract":"<p>Nitrogen availability critically influences biofilm development in drinking water distribution systems (DWDSs), yet the distinct impacts of ammonium (NH₄⁺) and nitrate (NO₃⁻) on biofilm and pathogen risks remain unclear. Using bench-scale annular reactors simulating DWDSs, we investigated how ammonium depletion/supplementation (AD/AS) and nitrate depletion/supplementation (ND/NS) regulate biofilm profiles over 28 days. Water chemistry analyses and functional annotation (KEGG, FAPROTAX) revealed that AS and NS increased total nitrogen retention (AS: 2.046–2.261 mg/L; NS: 10.625–11.800 mg/L) and biofilm prokaryotic cell abundance. AS enriched biosynthesis and virulence pathways (e.g., amino acid transport, energy production), while AD activated stress-response mechanisms (e.g., DNA repair, carbohydrate metabolism). NS favored Gram-negative bacteria, stress-tolerant taxa, and pathogenic genera (e.g., <i>Acinetobacter</i>, <i>Mycobacterium</i>), whereas ND upregulated organic nitrogen scavenging. Nitrogen supplementation preferentially enriched genera including WHO-listed critical-risk pathogens, which correlated strongly with inorganic nitrogen levels (Mantel r &gt; 0.5, <i>P</i> &lt; <i>0.05</i>). In contrast, genera including high/medium-risk pathogens exhibited increased relative abundance under depletion via organic nutrient reliance. These findings demonstrate that NH₄⁺ and NO₃⁻ differentially regulate biofilm pathogenicity: NH₄⁺ drives growth and virulence, while NO₃⁻ enhances structural resilience and redox flexibility. The study advocates targeted nitrogen management in DWDS to monitor and mitigate biofilm-associated health risks and for optimizing microbial stability.</p><p>• <i>Nitrogen species (NH₄⁺ and NO₃⁻) enrichment and availability elevated biofilm cell density and altered functional groups and metabolic processes, revealing strong coupling between nutrient availability and microbial proliferation in DWDSs.</i></p><p>• <i>Ammonium enrichment drove virulence- and metabolism-related functions, whereas nitrate promoted oxidative resilience and biofilm matrix stability.</i></p><p>• <i>Nitrogen limitation activated stress-adaptive and organic nitrogen scavenging pathways, reshaping community composition toward stress-tolerant, opportunistic taxa.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13905-y.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148350404","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"N-terminal lipidation enhances membrane interaction and antifungal activity of a Trematocine-derived decapeptide","authors":"Damiano Squitieri,&nbsp;Federica Massaro,&nbsp;Melinda Mariotti,&nbsp;Margherita Cacaci,&nbsp;Stefano Borocci,&nbsp;Fernando Porcelli,&nbsp;Francesco Buonocore,&nbsp;Francesca Bugli,&nbsp;Maurizio Sanguinetti","doi":"10.1007/s00253-026-13929-4","DOIUrl":"10.1007/s00253-026-13929-4","url":null,"abstract":"<p>Invasive fungal infections are a growing global health threat, driven by the limited availability of antifungal therapies and the rapid spread of multidrug-resistant yeasts. This study explored the effects of N-terminal lipidation on Trem-4, a decapeptide derived from the antimicrobial peptide Trematocine-HSK, to enhance its antifungal potential. Trem-4 was modified with caprylic (C8) and myristic (C14) fatty acid chains, generating two lipopeptides: Cap-T4 and Myr-T4. Biophysical analyses, including fluorescence-based assays and circular dichroism, showed that lipidation significantly improved peptide–membrane interactions compared to the non-lipidated form. The two derivatives displayed distinct behaviors, with differences in aggregation and membrane selectivity depending on acyl chain length. Antifungal activity was assessed against 60 clinically characterized yeast isolates, including <i>Candida</i> spp., <i>Candidozyma haemuli complex</i>, and <i>Cryptococcus neoformans</i>, under both planktonic and biofilm conditions. Both lipopeptides exhibited broad-spectrum activity, against clinically relevant species, with conserved activity against antifungal-resistant strains. Importantly, both compounds remained effective against mature biofilms, causing metabolic disruption and structural damage. Cap-T4 demonstrated consistent activity across species, while Myr-T4 showed enhanced potency against selected isolates, particularly <i>C. auris</i>. Biocompatibility evaluation revealed a concentration-dependent cytotoxicity and hemolysis, more pronounced for Myr-T4. However, both compounds were well tolerated in vivo in the <i>Galleria mellonella</i> model. Overall, Cap-T4 emerged as the most promising candidate for further preclinical development. These findings support N-terminal lipidation as an effective strategy to improve the antifungal efficacy of short antimicrobial peptides and highlight its potential for developing new treatments against drug-resistant fungal pathogens.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13929-4.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148350517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improving the reproducibility of mixing-time experiments in stirred single-use bioreactors","authors":"Isabelle Barth,&nbsp;Christian Holbeck,&nbsp;Charlotte Capitain,&nbsp;Tanja Schirmeister,&nbsp;Percy Kampeis","doi":"10.1007/s00253-026-13931-w","DOIUrl":"10.1007/s00253-026-13931-w","url":null,"abstract":"<p>Mixing time is a key metric for characterizing bioreactors, as it governs bulk homogenization and correlates with the underlying flow field and shear exposure. Having knowledge of turbulent flow regimes is particularly important for shear-sensitive (mammalian) cells, which are often cultivated in single-use systems. To characterize such systems in terms of their mixing-time behavior, colorimetric pH-shift assays can be employed. However, operator-dependent variability during the reagent addition often limits the comparability of such experiments. To address this limitation and enhance reproducibility, a compact linear actuator was developed that enables automation and, thus, standardization of the addition process. The device was designed using computer-aided design software, fabricated via 3D printing, and subsequently integrated into mixing-time experiments. It is compatible with both stirred single-use bioreactors and glass bioreactors. For automated experiments using the linear actuator, neither the mixing time nor the replicate variance increased in the single-use bioreactor relative to the glass bioreactor under non-aerated conditions, demonstrating strong comparability between the two systems. The use of the linear actuator increased measurement precision, as evidenced by a 13.6% reduction in the coefficient of variation. This improvement resulted from the injection profiles, which were consistent in their immersion depth, immersion angle, tracer volume, and injection speed. Accordingly, this approach provides a low-cost, plug-and-play method to improve the reproducibility of mixing-time experiments, thereby facilitating bioreactor characterization, technology transfer, and validation of computational fluid dynamics models.</p><p><i>Automated acid/base dosing improves reproducibility versus manual addition.</i></p><p><i>Variation of the local mixing is detectable through spatially resolved analysis.</i></p><p><i>Enables robust characterization of both stirred single-use and glass bioreactors.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13931-w.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148350480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of controlled fermentation by Lactiplantibacillus plantarum PBS067 on Vitis vinifera L. leaf extract","authors":"Marta Armari,&nbsp;Federica Cascella,&nbsp;Aurora Tritto,&nbsp;Farah Hamed,&nbsp;Shwetha Bangalore Renukaprasad,&nbsp;Ana Moreno Villena,&nbsp;Giuseppe Meca De Caro,&nbsp;Carlos Luz Mínguez,&nbsp;Cesar Francisco Trejo,&nbsp;Barbara Azzimonti","doi":"10.1007/s00253-026-13942-7","DOIUrl":"10.1007/s00253-026-13942-7","url":null,"abstract":"<p><i>Vitis vinifera</i> L. leaves represent one of the most phenolic-rich grapevine biomasses, exhibiting a broad spectrum of biological activities, including prebiotic and microbiota-modulating ones. However, as they are mainly regarded as agro-industrial waste, they are still an underexploited resource. Probiotic-driven fermentation may offer a promising strategy to enhance their bioactive potential while addressing this challenge. The aim of this study was to evaluate the effects of the in vitro fermentation of <i>Vitis vinifera</i> L. leaf extract (VVLE) by <i>Lactiplantibacillus plantarum</i> PBS067 in terms of phenolic content, antioxidant activity, lactic acid and protein content, and metabolomic profile. Our analysis shows the prebiotic activity of VVLE, as VVLE-supplemented medium allowed probiotic growth without toxic effects. Leaves fermentation led to a significant enrichment in lactic acid and the biotransformation of complex plant-secondary metabolites into smaller derivatives. However, VVLE antioxidant capacity slightly decreased after the fermentation, likely due to the phenolic conversion itself. These results highlight how controlled lacto-fermentation valorizes grapevine leaves, transforming them into sustainable upcycled bioactive products with potential applications within cosmetics, pharmaceuticals, nutraceuticals and complementary medical product sectors.</p><p>• <i>L. plantarum PBS067 thrives in medium supplemented with polyphenol-rich leaf extract</i></p><p>• <i>L. plantarum PBS067-driven fermentation modulates VVLE phytochemical profile</i></p><p>• <i>L. plantarum PBS067-driven fermentation valorizes viticulture by-products</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13942-7.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148350381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Poly(3-hydroxybutyrate) production from ricotta whey by Cupriavidus necator: effects of coffee oil and oxidative stress","authors":"Angela Longo,&nbsp;Andrea Torreggiani,&nbsp;Luca Sconosciuto,&nbsp;Michela Verni,&nbsp;Vito Emanuele Carofiglio,&nbsp;Domenico Centrone,&nbsp;Marianna Villano,&nbsp;Gaia Salvatori,&nbsp;Erica Pontonio,&nbsp;Marco Montemurro,&nbsp;Carlo G. Rizzello","doi":"10.1007/s00253-026-13933-8","DOIUrl":"10.1007/s00253-026-13933-8","url":null,"abstract":"<p>In this study, ricotta cheese exhausted whey (RCEW), an abundant dairy by-product, was evaluated as a low-cost substrate for poly(3-hydroxybutyrate) (PHB) production by <i>Cupriavidus necator</i> DSM 428 through an integrated process strategy combining substrate conditioning, enzymatic lactose hydrolysis, oxidative stress modulation, and cultivation under controlled bioreactor conditions. Shake-flask experiments demonstrated that RCEW supported efficient biomass formation (1.60 g L⁻¹) and high intracellular PHB accumulation (71.4% of dry cell weight, DCW), while supplementation with coffee oil extract increased biomass up to 4.41 g L⁻¹ and PHB titer to 2.87 g L⁻¹. Controlled oxidative stress further enhanced polymer accumulation, with hydrogen peroxide addition (5.0 mmol L⁻¹ at 24 h) promoting intracellular PHB contents up to 80.8% DCW. Cultivation in a stirred-tank bioreactor confirmed process robustness, and fed-batch cultivation resulted in PHB concentrations up to 3.85 g L⁻¹ (77.9% DCW) using a chlorine-free downstream recovery protocol. The recovered polymer consisted exclusively of 3-hydroxybutyrate units and exhibited a high–molecular weight (876 kDa). Overall, these results demonstrate the feasibility of producing high-quality PHB from RCEW using a non-recombinant <i>C. necator</i> strain, supporting the development of sustainable bioprocesses for dairy by-product valorization.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13933-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148337215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tandem triplicate A29L peptide elicits robust immune responses against monkeypox virus in mice","authors":"Chia-Yu Liang,&nbsp;Ting-Yu Chang,&nbsp;Pei-Hsuan Chen,&nbsp;Tai-Ling Chao,&nbsp;Sui-Yuan Chang,&nbsp;Shih-Chung Chang","doi":"10.1007/s00253-026-13936-5","DOIUrl":"10.1007/s00253-026-13936-5","url":null,"abstract":"<p>Monkeypox (Mpox) remains a significant public health concern due to the emergence of rapidly evolving strains and the suboptimal efficacy of existing smallpox vaccines. Recombinant subunit vaccines offer a promising and safe alternative. The A29L protein of the Mpox virus (MPXV) is essential for viral attachment and entry. The N-terminal residues 1–34 of A29L, which harbor the heparin-binding domain, are critical targets for neutralizing antibodies. Based on this, we hypothesized that this truncated 34-residue peptide of A29L could serve as a potent subunit vaccine candidate. To enhance immunogenicity, the recombinant antigen comprising three tandem repeats of the 34-residue peptide (designated as Tri(1–34)) was engineered. Following the immunization of BALB/c mice, Tri(1–34) elicited robust A29L-specific antibody titers comparable to those induced by the full-length protein, demonstrating equivalent immunogenicity. Furthermore, antisera collected from Tri(1–34)-immunized mice specifically recognized the A29L N-terminus and effectively neutralized MPXV infection in Vero E6 cells, as evidenced by the plaque reduction neutralization test (PRNT) assays. Immunization with Tri(1–34) also triggered strong Th1-biased cellular immune responses, characterized by significant interferon-gamma (IFN-γ) production. In conclusion, the engineered Tri(1–34) antigen represents a safe and effective subunit vaccine candidate, offering a strategic framework for future Mpox vaccine development.</p><p>• <i>Engineered Tri(1–34) targets the critical A29L N-terminus of Mpox virus</i></p><p>• <i>Tri(1–34) elicits potent neutralizing antibodies against Mpox virus</i></p><p>• <i>Tri(1–34) induces a robust Th1-biased response with high IFN-γ levels</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13936-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148337453","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cultivar and tree age jointly shape rhizosphere bacterial diversity and community assembly in Macadamia integrifolia","authors":"Xingze Li,&nbsp;Linan Kang,&nbsp;Chengmeng Zhang,&nbsp;Huiping Zeng,&nbsp;Guixiang Yu,&nbsp;Jiangmin Yang,&nbsp;Furong Li,&nbsp;Wenliang Cai,&nbsp;Yanhuang Feng,&nbsp;Haibo Wu","doi":"10.1007/s00253-026-13909-8","DOIUrl":"10.1007/s00253-026-13909-8","url":null,"abstract":"<p>This study investigates the regulatory effects of cultivar and tree age on the rhizosphere bacterial community of <i>Macadamia integrifolia</i>, aiming to provide theoretical support for targeted regulation of the rhizosphere microecosystem and high-quality, high-yield production in the industry. Using rhizosphere soil from 5-year-old and 8-year-old plants of five major cultivated cultivars as materials, the bacterial community characteristics were revealed through 16S rRNA gene high-throughput sequencing combined with R-based data analysis and visualization. The results show that the community is dominated by two phyla, Actinobacteria and Chloroflexi, with the core dominant genus being <i>Rhodoplanes</i>. Cultivar is the leading factor driving community differentiation, and the effect of tree age is cultivar-specific. Functionally, the community is dominated by amino acid, carbohydrate, and lipid metabolic pathways, which are well-adapted to the host plant’s growth demands; the co-occurrence network exhibits predominantly strong positive correlations and high modularity, suggesting synergistic adaptation and metabolic complementarity among dominant bacterial taxa. This study clarified that the rhizosphere bacterial community of <i>Macadamia integrifolia</i> is primarily regulated by cultivar, and has identified core functional taxa such as <i>Rhodoplanes</i>, laying a scientific foundation for the development of specialized microbial inoculants and the coordinated regulation of cultivar-microbe interactions.</p><p>• <i>Cultivar is the dominant driver of macadamia rhizosphere bacterial community, with tree age exerting cultivar-specific effects.</i></p><p>• <i>Rhodoplanes is the core dominant genus, and the bacterial community functions are dominated by metabolic pathways.</i></p><p>• <i>Macadamia rhizosphere bacterial co-occurrence network shows strong positive correlations and modularity, guiding microbial inoculant development.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13909-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148337689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A new Kayvirus vB_SauM-MUHD-1 combats Methicillin-resistant Staphylococcus aureus wound infections","authors":"Maha Kabil,&nbsp;Noha T. Abou El-Khier,&nbsp;Wafaa Mowafy,&nbsp;Abeer M. Abd El-Aziz,&nbsp;Mohamed Abdelmoteleb","doi":"10.1007/s00253-026-13898-8","DOIUrl":"10.1007/s00253-026-13898-8","url":null,"abstract":"<p>Methicillin-resistant <i>Staphylococcus aureus</i> (MRSA), a leading cause of chronic and post-surgical wound infections, is a hard-to-treat pathogen. In this study, we isolated and characterized a lytic MRSA bacteriophage, vB_SauM-MUHD-1, and evaluated its therapeutic potential in a murine wound infection model. Among 107 clinical wound samples, MRSA was reported in 48.6% of cases. Phage vB_SauM-MUHD-1, isolated from sewage, demonstrated lytic activity against 70.6% of the tested MRSA isolates. The phage exhibited efficient replication kinetics and remained stable under physiologically relevant conditions. Whole-genome sequencing identified a ~ 134 kb dsDNA genome (~ 30.45% GC) lacking detectable lysogeny-associated, virulence, or antimicrobial-resistance genes. In a BALB/c excisional wound model infected with MRSA, topical phage treatment significantly reduced bacterial burden, accelerated wound closure, and improved clinical severity scores compared to untreated controls, performing comparably to phage-linezolid combination therapy and outperforming linezolid monotherapy in bacterial clearance. These findings support that our phage vB_SauM-MUHD-1 has potential for treating MRSA-infected wounds and should be further investigated for efficacy in more challenging chronic or biofilm-rich wound environments.</p><p>• <i>This study provides a newly kayvirus, strictly lytic anti-MRSA phage vB_SauM-MUHD-1.</i></p><p>• <i>Phage exhibited favorable replication kinetics, physical stability and genomic safety.</i></p><p>• <i>Topical phage therapy reduced bacterial burden and accelerated wound healing.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13898-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148323720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human papillomavirus 18 E6-specific Affibody inhibits cervical cancer cell proliferation and induces apoptosis","authors":"Haiyan Dong,&nbsp;Jing Zhang,&nbsp;Le Wu,&nbsp;Xi Zhang,&nbsp;Jun Chen,&nbsp;Wangqi Du,&nbsp;Saidu Kamara,&nbsp;Shanli Zhu,&nbsp;Lifang Zhang","doi":"10.1007/s00253-026-13903-0","DOIUrl":"10.1007/s00253-026-13903-0","url":null,"abstract":"<p>Human papillomavirus (HPV) 18 E6 oncoprotein drives cervical carcinogenesis by degrading p53, enabling uncontrolled cell proliferation. In this study, we developed novel trans-activator of transcription (TAT)-conjugated Affibody molecules (TAT-Z_HPV18E6) targeting HPV18 E6 for therapeutic applications. High-affinity Affibody variants were screened by phage display using recombinant HPV18 E6 expressed in <i>Escherichia coli</i>. Three candidates (TAT-Z_HPV18E6: 4, 59, 352) exhibited high binding affinity, with equilibrium dissociation constant (<i>K</i>_D) ranging from 10⁻⁶ to 10⁻⁴ M, with TAT-Z_HPV18E6: 59 showing superior specificity for native HPV18 E6 in HeLa229 cells, as validated by surface plasmon resonance (SPR), enzyme-linked immunosorbent assay (ELISA), and immuno-fluorescence. In vivo near-infrared fluorescence imaging of DyLight 755-labeled TAT-Z_HPV18E6: 59 in tumor-bearing mice demonstrated rapid tumor accumulation (peak at 2 h) and prolonged retention (&gt; 12 h). Mechanistically, TAT-Z_HPV18E6: 59 restored p53 stability and upregulated pro-apoptotic factors (Bax, PUMA) and cell cycle regulator p21. Notably, combined treatment with TAT-Z_HPV18E6: 59 and E7-targeting TAT-Z_HPV18E7: 228 combinatorial enhanced apoptosis and suppressed HeLa229 proliferation, as confirmed by CCK-8 and clonogenic assays. These results demonstrate the utility of TAT-Z_HPV18E6 Affibody molecules as targeted agents, highlighting combinatorial E6/E7 targeting as a potent strategy for HPV18-driven cervical cancer therapy.</p><p>• <i>TAT-Z</i>_HPV18E6:<i> 59 achieved high affinity for HPV18 E6, enabling precise molecular targeting.</i></p><p>• <i>Demonstrates dual utility for in vivo imaging and p53-dependent tumor suppression.</i></p><p>• <i>Dual targeting of E6/E7 yields synergistic therapeutic efficacy in cervical cancer models.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13903-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148337254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Redox-based dual-substrate fermentation enhances sustainable production of 3-hydroxypropionic acid by recombinant E. coli","authors":"Kala Baskar,&nbsp;Vishnu Prasad J,&nbsp;Guhan Jayaraman","doi":"10.1007/s00253-026-13926-7","DOIUrl":"10.1007/s00253-026-13926-7","url":null,"abstract":"<p>3-Hydroxypropionic acid (3-HP) is an important platform chemical for a circular bioeconomy since it is a precursor for the production of several high-value derivatives such as acrylic acid as well as biodegradable plastics. <i>Escherichia coli</i> is a promising host for 3-HP biosynthesis due to its well-characterized metabolic network and ease of genetic manipulation. In <i>E. coli</i>, 3-HP is produced from glycerol via a two-step pathway involving glycerol dehydratase and an NAD⁺-dependent aldehyde dehydrogenase. However, a major limitation in this process is redox imbalance, as both glycerol catabolism and 3-HP biosynthesis generate excess NADH, thereby restricting NAD⁺ availability and limiting product formation. In this study, we systematically evaluated the influence of cosubstrate selection to enhance 3-HP production by decoupling biomass formation from product synthesis while achieving redox balance. An engineered <i>E. coli</i> strain, with knockouts of glycerol kinase (<i>glpK</i>) and glycerol dehydrogenase (<i>gldA</i>) to prevent glycerol catabolism, was employed to redirect glycerol flux exclusively toward 3-HP biosynthesis. A secondary carbon source was supplied to support cell growth and promote NAD⁺ regeneration. Six different sugars were evaluated as growth substrates to assess their impact on redox balance and 3-HP production. Among the tested secondary substrates, sucrose and xylose enabled high 3-HP titers of 61 and 50 g/L, respectively, significantly surpassing the 30 g/L titer obtained using glycerol as the sole carbon source. The strategy was successfully extended to renewable feedstocks such as molasses and pretreated lignocellulosic liquor hydrolysate in combination with crude glycerol, achieving 3-HP titers in the range of ~ 35–40 g/L.</p><p>• <i>Redox imbalance limits glycerol-based 3-HP biosynthesis in E. coli.</i></p><p>• <i>Redox-optimized dual-substrate feeding enhances 3-HP titer, yield and productivity.</i></p><p>• <i>Validation with renewable substrates provides feasibility for a circular bioeconomy.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"110 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2026-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-026-13926-7.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148337198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}