Journal of biotechnology最新文献

{"title":"Novel polygalacturonase PG-BG31 prevents biofilm formation and increases antibiotic efficacy against catheter-associated Escherichia coli","authors":"Marija Atanaskovic ,&nbsp;Andjela Dilparic ,&nbsp;Mario Zlatović ,&nbsp;Lidija Senerovic","doi":"10.1016/j.jbiotec.2025.07.011","DOIUrl":"10.1016/j.jbiotec.2025.07.011","url":null,"abstract":"<div><div><em>Escherichia coli</em> is a leading cause of urinary tract infections, which are particularly problematic in catheterized patients due to the formation of biofilms that are resistant to standard treatments. Here we investigated a novel polygalacturonase (PG-BG31) from <em>Pedobacter</em> sp. BG31 and its anti-biofilm properties against <em>E. coli</em> isolates originating from the urine of UTI patients. The enzyme was heterologously expressed and biochemically characterized using polygalacturonic acid as substrate. The enzyme showed strong biofilm inhibition with BIC50 values between 2 and 5 µg/mL, which varied depending on the <em>E. coli</em> strain. It outperformed enzymes commonly used in <em>E. coli</em> biofilm studies, including proteinase K, DNase and cellulase. When used as a pretreatment, it significantly improved the efficacy of ciprofloxacin and trimethoprim by reducing biofilm formation by 4 log (99.99 %) at lower antibiotic concentrations. Polygalacturonase PG-BG31 works optimally at temperatures of 25 °C - 42 °C and a slightly acidic pH value (pH 5.0 – 6.0), which corresponds to the environment in urine. In addition, the enzyme showed no toxicity to cells in culture or <em>C. elegans</em>. These results suggest that polygalacturonase has the potential for the development of anti-biofilm strategies for catheter-related urinary tract infections.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 179-189"},"PeriodicalIF":4.1,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144662493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced exogenous RNAi by loop-ended double-stranded RNA in plants","authors":"Neil A. Smith ,&nbsp;Max Corral ,&nbsp;Canwei Shu ,&nbsp;Jiaozi Chen-Wang ,&nbsp;Daai Zhang ,&nbsp;Chengcheng Zhong ,&nbsp;Karam Singh ,&nbsp;Craig C. Wood ,&nbsp;Ling-Ling Gao ,&nbsp;Jonathan P. Anderson ,&nbsp;Ian K. Greaves ,&nbsp;Ming-Bo Wang","doi":"10.1016/j.jbiotec.2025.07.010","DOIUrl":"10.1016/j.jbiotec.2025.07.010","url":null,"abstract":"<div><div>Exogenous RNA interference (exoRNAi) has emerged as a promising tool for gene silencing in plants, yet a robust and efficient exoRNAi technology remains elusive. Here we evaluated a Loop-ended dsRNA (ledRNA) design for enhancing exoRNAi efficacy. ledRNA has a dumbbell-like structure, consisting of a long dsRNA stem flanked by single-stranded loops, with a nick site at the dsRNA stem. When topically applied to leaves, cotyledons, meristems or roots, the ledRNA accumulated at higher levels than traditional hairpin RNA (hpRNA) both in the treated tissues (leaf, cotyledon, roots) and untreated tissues (adjacent leaf areas and remote cotyledons and roots). This suggests that ledRNA possesses greater stability and more efficient uptake and movement in plants than the traditional RNAi molecules. Topically delivered ledRNA induced stronger silencing of the endogenous <em>FAD2</em> gene in <em>N. benthamiana</em> leaves, leading to increased accumulation of monounsaturated fatty acids. Beyond plants, ledRNA also exhibited RNAi activity in diverse kingdoms of life. When introduced into cultures of the fungus <em>Fusarium oxysporum</em>, ledRNA effectively and specifically downregulated expression of the <em>FoCYP51A</em>, <em>FoCYP51B, FoCYP51C, FoSDHB</em>, <em>FoSDHC</em> and <em>FoSDHD</em> genes in the fungal cells. Additionally, oral ingestion of ledRNA targeting the critical <em>MpRack-1</em> gene by green peach aphids (<em>Myzus persicae</em>) significantly reduced their fecundity, leading to a 75 % decrease in progeny production. These findings highlight ledRNA as a promising tool for enhancing the efficacy of exoRNAi-based manipulation of gene expression.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 198-210"},"PeriodicalIF":4.1,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144649575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The CRISPR Cas patent files, part 3: Prime Editing and integrase-based variants","authors":"Ulrich Storz","doi":"10.1016/j.jbiotec.2025.07.006","DOIUrl":"10.1016/j.jbiotec.2025.07.006","url":null,"abstract":"<div><div>The epic patent disputes regarding CRISPR Cas9 and, to a lesser extent, CRISPR Cas12a, have somehow overcast a new patent debate that is materializing at the horizon, namely the one that relates to Prime Editing and its integrase-based siblings. While these techniques offer great promise for the precise integration of large DNA stretches into a host genome, it appears that a new chain of dependencies is about to unfold that makes the establishment of Freedom to Operate complicated for interested parties. (Arnold, 2024).</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 147-153"},"PeriodicalIF":4.1,"publicationDate":"2025-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144633162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acid-triggered ultralong hydrogen release from AB-loaded hollow mesoporous silica nanoparticles enhances salt tolerance in sweetpotato","authors":"Nan Xia ,&nbsp;Zheng Zhuang ,&nbsp;Qi Wang ,&nbsp;Zhiyuan Pan ,&nbsp;Yicheng Yu ,&nbsp;Zongyun Li ,&nbsp;Jian Sun ,&nbsp;Yanjuan Li","doi":"10.1016/j.jbiotec.2025.07.008","DOIUrl":"10.1016/j.jbiotec.2025.07.008","url":null,"abstract":"<div><div>Soil salinization severely restricts crop growth and productivity worldwide. Hydrogen (H₂) has demonstrated considerable potential in enhancing plant stress tolerance. However, its agricultural application is limited by the rapid dissipation of H₂ in traditional electrolysis-generated hydrogen-rich water (HRW). In this work, we synthesized a nanoscale H₂-releasing material (AB@HMSN) by encapsulating ammonia borane (AB) into hollow mesoporous silica nanoparticles (HMSN), achieving ultrahigh AB loading capacity (811 mg·g⁻¹ HMSN) and acid-responsive sustained H₂ release behavior in solution. The H₂ release duration reached 75 h, significantly exceeding that of conventional HRW. When applied to salt-stressed (150 mM NaCl) sweetpotato (<em>Ipomoea batatas</em> (L.) Lam) seedlings, 16 mg·L⁻¹ AB@HMSN significantly improved chlorophyll content, with chlorophyll a, chlorophyll b, and total chlorophyll increasing by 80 %, 100 %, and 88 %, respectively. Photosynthetic performance was markedly enhanced, showing increases of 53 %, 26 %, and 24 % in photosynthetic rate (Pn), stomatal conductance (Gs), and transpiration rate (Tr), respectively. Additionally, AB@HMSN treatment strengthened antioxidant defenses by elevating the activities of key enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), while reducing the accumulation of reactive oxygen species (H₂O₂ and ·O^2-). Mechanistically, AB@HMSN induced endogenous melatonin (MT) production, which activated sodium-proton (Na⁺/H⁺) antiporters and plasma membrane proton-pumping ATPase (PM H⁺-ATPase) activity. This process restored ion homeostasis by facilitating Na⁺ efflux, K⁺ influx, and H⁺ influx, thereby enhancing salt tolerance. These findings not only advance our understanding of hydrogen-mediated stress tolerance but also offer a practical nanomaterial-based strategy for sustainable agriculture in saline environments.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 105-114"},"PeriodicalIF":4.1,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144604306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Heterologous expression of L-lactate dehydrogenase in Chlorella pyrenoidosa: A beginning for poly(L-lactic acid) biosynthesis from CO2 by eukaryotic microalgae","authors":"Xu Zhang ,&nbsp;Xinxin Meng ,&nbsp;Jiaxin Liang ,&nbsp;Dejing Kong ,&nbsp;Yike Qi ,&nbsp;Di Cai ,&nbsp;Bin Wang ,&nbsp;Yong Wang","doi":"10.1016/j.jbiotec.2025.07.009","DOIUrl":"10.1016/j.jbiotec.2025.07.009","url":null,"abstract":"<div><div>Lactic acid is a monomer of poly(lactic acid), and the use of sugar-based raw materials in conventional lactic acid production poses a risk to food security. CO₂ is an abundant carbon resource, and microalgae are crucial for the development of third-generation bio-manufacturing through CO₂ utilization. In this study, the transformation efficiency of <em>Chlorella pyrenoidosa</em> (<em>C. pyrenoidosa</em>) was investigated during <em>Agrobacterium</em>-mediated transformation, and an efficient transformation system was established. L-lactate dehydrogenases (LLDHs) from <em>Bos taurus</em>, <em>Lactobacillus plantarum</em>, and <em>Bacillus coagulans</em> were successfully integrated into <em>C. pyrenoidosa</em>, and the expression of the target genes was confirmed through RT-PCR and enzyme activity assays. The engineered strains expressing <em>BCLLDH</em>, <em>LPLLDH</em>, and <em>BTLLDH</em> produced L-lactic acid titers of 76.64 mg/L, 153.18 mg/L, and 98.21 mg/L, respectively, and their transgenic stability was verified. Homology modeling of the three LLDHs was performed, and molecular docking with pyruvate was conducted to investigate the interaction mechanism. Moreover, binding energy and interaction network analyses supported the obtained enzyme activity results. Ultimately, L-lactic acid biosynthesis was achieved in <em>C. pyrenoidosa</em>, offering a potential pathway for eukaryotic microalgae to synthesize poly(L-lactic acid) based on the generated precursor.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 136-146"},"PeriodicalIF":4.1,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144618051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional nucleic acid-powered hybrid nanocarriers for synchronized targeted delivery of dual-solubility therapeutics","authors":"Xin Liu ,&nbsp;Mengqiu Sun ,&nbsp;Lu Dai ,&nbsp;Yusheng Li ,&nbsp;Rui Song ,&nbsp;Hong Bian ,&nbsp;Hao Zhang","doi":"10.1016/j.jbiotec.2025.07.002","DOIUrl":"10.1016/j.jbiotec.2025.07.002","url":null,"abstract":"<div><div>Combination chemotherapy presents a promising approach to overcome multidrug resistance in cancer treatment. However, conventional delivery systems struggle to achieve synchronized co-delivery of hydrophilic and hydrophobic therapeutics with tumor-specific targeting and controlled release. Here, we developed a multifunctional nanocarrier integrating poly (lactic-co-glycolic acid) nanoparticles with polydopamine and programmable nucleic acids. Within this system, the hydrophobic core encapsulates lipophilic drugs, while the surface nucleic acid architecture enables hydrophilic drug loading via molecular intercalation. Furthermore, tumor-targeting aptamers guide selective cellular uptake, and a tumor microenvironment-responsive mechanism ensures spatiotemporally controlled drug release. As demonstrated experimentally, this unified platform demonstrates promising antitumor activity with potential advantages in drug delivery efficiency. By combining targeted delivery, differential drug loading, and stimuli-triggered release, the design establishes a modular framework for precision combination therapies, thereby advancing adaptable nanomedicine in oncology.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 127-135"},"PeriodicalIF":4.1,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144604339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel approaches to engineering autolysis: A scalable alternative for recovery of bioproducts.","authors":"Kyle Gaius Jonsson,&nbsp;Darren Nesbeth,&nbsp;Eli Keshavarz-Moore","doi":"10.1016/j.jbiotec.2025.07.007","DOIUrl":"10.1016/j.jbiotec.2025.07.007","url":null,"abstract":"<div><div>For intracellularly expressed bioproducts, cell disruption is an essential unit operation in the bioprocess. Traditional cell lysis techniques, such as homogenisation or chemical lysis, may result in product damage due to the shear forces or the harsh chemicals used, respectively. Autolysis, or the expression of lytic proteins to trigger self-lysis, presents as an alternative approach for intracellular bioproduct release. However, challenges of using autolysis in bioprocessing have hindered the wide adoption of the technique. There are three main issues related to autolysis: leaky gene expression leading to reduced performance in fermentation, low lysis efficiencies compared to traditional methods, and a lack of consideration for scale-up in the current literature. This review summarises these challenges and discusses the potential solutions.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 115-126"},"PeriodicalIF":4.1,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144604305","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microwave-assisted biogenic synthesis of pure copper oxide nanoparticles by Lepidium sativum L. leaves extract: Characterization and evaluation of antibacterial activity","authors":"Ahmed A. Ibrahim","doi":"10.1016/j.jbiotec.2025.07.003","DOIUrl":"10.1016/j.jbiotec.2025.07.003","url":null,"abstract":"<div><div>This study delineates a sustainable and environmentally compatible route for the biogenic synthesis of copper oxide nanoparticles (CuO-NPs) utilizing the leaf extract of <em>Lepidium sativum L</em>. as a biocompatible reducing agent, employing microwave technology, alongside an evaluation of their antibacterial activity and biofilm formation against <em>Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli,</em> and <em>Streptococcus mutans</em>. The size, shape, crystalline structure, and stability of the synthesized CuO-NPs were determined by energy dispersive X-ray (EDX), scanning electron microscopy (SEM), X-ray diffraction (XRD), infrared (IR) and ultraviolet-visible (UV-Visible), dynamic light scattering (DLS), and X-ray photoelectron spectroscopy (XPS). The stability of the synthesized CuO-NPs exhibited exceptional stability within 90 days. XRD confirmed that the synthesized CuO-NPs had a spherical morphology, with an average particle diameter of 30 nm. Furthermore, CuO-NPs displayed strong antibacterial activity against the bacterial strains, as well as significant prevention of biofilm formation. This study's findings confirmed the successful synthesis of an effective, economically viable, and eco-friendly synthesis method for CuO-NPs. The results reveal their substantial antibacterial activities, indicating their use as potential alternatives to standard antimicrobial therapies, with major advantages in terms of cost and biocompatibility.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 82-90"},"PeriodicalIF":4.1,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144583999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of 3D-printed acrylonitrile butadiene styrene and polylactic acid carrier-beads for Gluconobacter oxydans immobilization","authors":"Ricardo Gonzalo Ramírez Brenes ,&nbsp;Isabella Maria Tenório Soares Santos ,&nbsp;Livia da Silva Chaves de Carvalho ,&nbsp;Ninoska Isabel Bojorge Ramírez ,&nbsp;Nei Pereira Jr.","doi":"10.1016/j.jbiotec.2025.07.001","DOIUrl":"10.1016/j.jbiotec.2025.07.001","url":null,"abstract":"<div><div>Bioprocesses play a crucial role in the production of biotechnological compounds, and 3D printing offers innovative solutions, including the creation of beads for cell immobilization. This technology enables the development of customized structures with various geometries and properties (shape, size, porosity, density, and roughness). This study produced carrier beads using fused filament fabrication with two different thermopolymers: acrylonitrile butadiene styrene (ABS) and polylactic acid (PLA). The carrier beads were printed with three distinct millipore geometries—hexagonal, square, and triangular—to evaluate immobilizing <em>Gluconobacter oxydans</em>' efficiency and investigate how immobilization varies based on the material type and millipore geometry. ABS exhibited greater surface roughness compared to PLA, which enhanced cell immobilization. The highest cell immobilization efficiencies were achieved with the ABS beads, with the hexagonal millipore geometry demonstrating the best efficiency at 84.7 %. Determining mechanical parameters, including storage and loss modulus, is essential for ensuring the durability of the beads in industrial processes. The glass transition temperatures for ABS and PLA were found to be 105 °C and 60 °C to 65 °C, respectively. Thus, 3D printing stands out for its versatility in material selection and geometry, optimizing the performance of bioprocesses.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 154-167"},"PeriodicalIF":4.1,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144575506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient genome engineering in Agrobacterium tumefaciens C58 using recombineering assisted by CRISPR/Cas9","authors":"Shiqing Gao ,&nbsp;Xiuling Wang ,&nbsp;Runyu Yang,&nbsp;Zhilong Bian,&nbsp;Daojing Yong,&nbsp;Shouying Xu,&nbsp;Liujie Huo,&nbsp;Qiang Tu,&nbsp;Youming Zhang,&nbsp;Jun Fu,&nbsp;Ruijuan Li","doi":"10.1016/j.jbiotec.2025.07.005","DOIUrl":"10.1016/j.jbiotec.2025.07.005","url":null,"abstract":"<div><div>Recombineering, a technique derived from phage-encoded homologous recombination, has emerged as a vital approach for bacterial genome engineering. <em>Agrobacterium tumefaciens</em> is extensively utilized to transfer DNA into the host plant genomes. To facilitate the transformation of various plant species, particularly those of considerable economic value, genetic modifications of <em>Agrobacterium</em> strains are essential. Our previous studies established an <em>Agrobacterium</em>-specific phage-encoded homologous recombination system for <em>Agrobacterium</em> species. Yet, recent investigations have indicated that there is a substantial variability in the recombination efficiency of these recombineering systems for gene editing across different genome loci in <em>A. tumefaciens</em>. In this work, we present the development of an efficient genome engineering tool for <em>A. tumefaciens</em> by integrating recombineering with CRISPR/Cas9 technology. Initially, we found that lengthening the homology arms significantly enhanced genome editing efficiency. Nevertheless, at certain genomic sites, even when the length of the homology arms was increased, the editing efficiency remained suboptimal. Subsequently, combination of the <em>Agrobacterium</em>-specific recombineering system with the CRISPR/Cas9 system markedly enhanced the genome engineering efficiency. This study offers an enhanced and efficient genome engineering tool for <em>A. tumefaciens</em>, which could potentially be applied to other species within the <em>Agrobacterium</em> genus.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"406 ","pages":"Pages 99-104"},"PeriodicalIF":4.1,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144575507","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}