Microbial Biotechnology最新文献

{"title":"Characterisation of a Novel Jumbo Lytic Aeromonas dhakensis Bacteriophage P19 and Its Endolysin","authors":"Farhat Ansari,&nbsp;Vandan Nagar","doi":"10.1111/1751-7915.70289","DOIUrl":"10.1111/1751-7915.70289","url":null,"abstract":"<p><i>Aeromonas dhakensis</i> is an emerging multidrug-resistant (MDR) bacterial pathogen that presents significant threats to public health and aquaculture. This study reports the isolation and comprehensive characterisation of jumbo bacteriophage P19. It is a lytic phage with a long, contractile tail and is placed in the genus <i>Ceceduovirus</i> of the family <i>Straboviridae</i>. P19 possesses a large genome typical of jumbo phages, which likely contributes to its enhanced replication efficiency and adaptability. P19 exhibited strong lytic activity with a high efficiency of plating (EOP) against multiple <i>A. dhakensis</i> strains, a burst size of approximately 50 plaque-forming units (PFU) per cell, and a latent period of 30 min. It remained highly stable across pH 6–10 and temperatures up to 60°C. Genomic analysis revealed a 228 kb genome with 414 open reading frames (ORFs) and absence of genes related to lysogeny, antibiotic resistance or virulence, indicating its potential for safe therapeutic and biocontrol applications. ORF_358 encodes a stable and soluble T4-type lysozyme endolysin (molecular weight 18.8 kDa) belonging to the glycoside hydrolase family 24. Molecular docking analysis demonstrated strong binding affinity (−6.4 kcal/mol) between P19_358 and the peptidoglycan NAG-NAM dimer. Recombinant P19_358 endolysin exhibited 99.98% relative lytic activity against Gram-positive bacterium <i>Bacillus subtilis</i> at a concentration of 75 μg/mL without EDTA pre-treatment; whereas, it showed broad-spectrum antibacterial activity against several Gram-negative pathogens, including <i>A. dhakensis</i>, <i>Shigella flexneri, Salmonella Typhimurium, Pseudomonas aeruginosa</i> and <i>Escherichia coli,</i> with relative activity ranging from 71% to 97% following pre-treatment with 100 mM EDTA. These findings collectively indicate that phage P19 and its endolysin P19_358 possess potent lytic activity and favourable stability profiles, supporting their development as promising antimicrobial agents against MDR Gram-negative pathogens.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 1","pages":""},"PeriodicalIF":5.2,"publicationDate":"2026-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12771599/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145905353","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cyanobacterial Extracellular Vesicles as Protein Carriers: Towards Fish Vaccination","authors":"Jorge Matinha-Cardoso,&nbsp;Gabriela Gonçalves,&nbsp;Filipe Coutinho,&nbsp;Steeve Lima,&nbsp;Lourenço Bonneville,&nbsp;Mónica Serrano,&nbsp;Paula Tamagnini,&nbsp;Aires Oliva-Teles,&nbsp;Ana Couto,&nbsp;Cláudia R. Serra,&nbsp;Paulo Oliveira","doi":"10.1111/1751-7915.70294","DOIUrl":"10.1111/1751-7915.70294","url":null,"abstract":"<p>Fish aquaculture faces significant economic losses from disease outbreaks. Vaccination is the most effective prevention strategy, and bacterial extracellular vesicles (EVs) show promise as vaccine platforms due to their strong immuno-stimulating properties. However, the application of EVs derived from pathogenic bacteria is limited by toxicity risks and production challenges. Alternatively, genetic engineering of non-pathogenic microorganisms is being explored to produce tailored EVs to deliver antigens and serve as carriers of therapeutic proteins. Recently, we have engineered the model cyanobacterium <i>Synechocystis</i> sp. PCC 6803 for the expression of the reporter green fluorescent protein (sfGFP) and its targeting to EVs. Here, taking advantage of the <i>Synechocystis</i> sfGFP-loaded EVs, the stability of vesicles and their cargo was evaluated in the long term when stored under different temperature conditions and after freeze-drying. The possibility of using <i>Synechocystis</i> EVs as a tool for eliciting specific/adaptive immune responses was assessed in European seabass, a high commercial value fish, by following the amount of total and sfGFP-specific immunoglobulins produced after immunisation through injection. <i>Synechocystis</i> EVs were shown to be resilient nanostructures that can induce specific immune responses in fish with additional adjuvant features. This represents a biotechnological breakthrough towards a novel antigen-carrier platform for sustainable fish-pathogen control.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 1","pages":""},"PeriodicalIF":5.2,"publicationDate":"2026-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://enviromicro-journals.onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.70294","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145888380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to “Combination of an Engineered Lactococcus lactis Expressing CXCL12 With Light-Emitting Diode Yellow Light as a Treatment for Scalded Skin in Mice”","authors":"","doi":"10.1111/1751-7915.70296","DOIUrl":"10.1111/1751-7915.70296","url":null,"abstract":"<p>Zhao, X., S. Li, J. Ding, J. Wei, P. Tian, H. Wei, and T. Chen. 2021. “Combination of an Engineered <i>Lactococcus lactis</i> Expressing CXCL12 With Light-Emitting Diode Yellow Light as a Treatment for Scalded Skin in Mice.” <i>Microbial Biotechnology</i> 14, no. 5: 2090–2100.</p><p>In Figure 2A, two unintentional image issues were identified: one is an unintentional reuse of images from the same HE section; the other is an inadvertent swapping of image positions representing two different time points within the same experimental group. These have been corrected in the version of Figure 2 shown below. It is important to note that these corrections do not affect the core conclusions of this study in any way.</p><p>We apologize for this error.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 1","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12743141/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145843380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microbial Community Restructuring Enhances Composting Efficiency: Synergistic Roles of Thermal Cycling and Fungal Inoculants (Fomes lignosus and Penicillium glabrum) in Metabolic Adaptation","authors":"Yukun Chen,&nbsp;Xiaofang Gong,&nbsp;Xiaobin Xiong,&nbsp;Gangjin Liu,&nbsp;Zhiye Wang,&nbsp;Ying Zhu","doi":"10.1111/1751-7915.70290","DOIUrl":"10.1111/1751-7915.70290","url":null,"abstract":"<p>This study demonstrates that synergistic integration of thermal cycling (28°C–58°C) and fungal inoculants (<i>Fomes lignosus</i>, <i>Penicillium glabrum</i>) enhances humification in cattle manure composting by restructuring microbial communities toward metabolic adaptation. Through a temperature-phased aerobic system, both inoculants significantly improved carbon conversion efficiency, with <i>F. lignosus</i> (B) and <i>P. glabrum</i> (G) increasing total organic matter by 4.71% and 3.42% (vs. control), humic acid content by 4.58-fold and 2.35-fold, and FDA hydrolase activity by 3.28-fold and 1.22-fold, respectively, confirming improved humification and nutrient cycling. Temperature–inoculant synergy drove functional differentiation. Respiratory profiling revealed that <i>P. glabrum</i> enhanced oxygen consumption by 1.3-fold during the early thermophilic phase (0–168 h at 58°C). Subsequently, temperature-induced respiration hierarchies (control &gt; B &gt; G) converged over time with microbial domestication. High-throughput sequencing and network analyses revealed that temperature–inoculant synergy reshaped the microbiome into simplified consortia, which comprise seven dominant bacterial phyla (e.g., <i>Firmicutes</i>, <i>Actinobacteriota</i>) and three dominant fungal phyla (e.g., <i>Ascomycota</i>), with marked functional differentiation characteristics. <i>P. glabrum</i> selectively enriched humification-related taxa, providing regulatory strategies for enhanced carbon stabilization; whereas <i>F. lignosus</i> favoured lignocellulose-degrading communities, optimising substrate valorisation efficiency. This strategy establishes a targeted microbial framework for optimising the resource utilisation efficiency of lignocellulosic waste within fermentation systems, thereby contributing to circular bioeconomy goals in sustainable organic waste management.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 12","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12742449/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145831734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Betulinic Acid Eradicates Implant-Associated Infections by Disrupting the S. aureus Biofilm Matrix and Potentiating Host Immune Clearance","authors":"Dongbin Guo,&nbsp;Ye Tao,&nbsp;Luanbiao Sun,&nbsp;Xinyao Liu,&nbsp;Yuan Gao,&nbsp;Peitong Jiang,&nbsp;Han Gao,&nbsp;Bingmei Wang,&nbsp;Li Wang","doi":"10.1111/1751-7915.70293","DOIUrl":"10.1111/1751-7915.70293","url":null,"abstract":"<p><i>Staphylococcus aureus</i> biofilms are major contributors to chronic and recurrent infections due to their intrinsic tolerance to antibiotics and host immune clearance, highlighting the urgent need for safe and effective antibiofilm strategies. This study evaluated the inhibitory effects and underlying mechanisms of betulinic acid (BA), the principal active constituent of the traditional Chinese medicine <i>Liquidambaris fructus</i>, against <i>S. aureus</i> biofilms. <i>In vitro</i> assays demonstrated that the minimum biofilm inhibitory concentration (MBIC) of BA was 32 μg/mL, which was markedly lower than its minimum inhibitory concentration (MIC, 512 μg/mL), indicating preferential activity against biofilm formation. Serial passage experiments revealed no detectable induction of drug resistance. Mechanistic studies revealed that BA suppressed early biofilm adhesion and aggregation, downregulated the expression of adhesion-related genes (<i>clfA</i>, <i>clfB</i>, <i>fnbpA</i> and <i>fnbpB</i>), and reduced the production of extracellular polysaccharide (EPS) and extracellular DNA (eDNA). BA further disrupted mature biofilm architecture, promoted macrophage infiltration, enhanced bacterial clearance and attenuated the expression of immune evasion factors (<i>scin</i>, <i>chip</i>, <i>lukE</i> and <i>nuc</i>). <i>In vivo</i>, BA significantly alleviated implant-associated infections, mitigated local inflammatory responses and facilitated tissue repair. Collectively, these findings reveal that BA inhibits <i>S. aureus</i> biofilms through multiple coordinated mechanisms, with a low propensity for resistance development and favourable biosafety, supporting its potential as a promising lead compound for the development of novel antibiofilm therapeutics.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"19 1","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12743172/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145843420","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 Modified Method Incorporating Multiplex PCR Reveals Fusobacterium Prevalence in Southern Chinese Population and Its Correlations in Cancers","authors":"Tingting Shen,&nbsp;Jiarui Liang,&nbsp;Xuyu Li,&nbsp;Xiaoxie Xu,&nbsp;Liqiong Li,&nbsp;Yinjuan Xu,&nbsp;Shanshui Zeng,&nbsp;Bingyu Li,&nbsp;Hui Li,&nbsp;Mengyao Hu,&nbsp;Lang Zhou,&nbsp;Siqi Yan,&nbsp;Ya Zhang,&nbsp;Ziwei Zhou,&nbsp;Huaaishi Liang,&nbsp;Shulei Chen,&nbsp;Zhikun Liang,&nbsp;Congrong Wang,&nbsp;Hongwei Zhou,&nbsp;Dongxin Zhang","doi":"10.1111/1751-7915.70292","DOIUrl":"10.1111/1751-7915.70292","url":null,"abstract":"<p><i>Fusobacterium</i>, a gram-negative anaerobic bacillus in mouth, gastrointestinal tract and elsewhere, has long been considered as opportunistic pathogen. Increasing evidence indicate the association of <i>Fusobacterium</i> with human diseases, especially cancer. However, previous studies demonstrated contradictory prevalent features of <i>Fusobacterium</i> species in normal and patient population. To address this dissonance, we developed a high-precision multiplex PCR assay that allows concurrent identification of five species of <i>Fusobacterium</i> (<i>F. nucleatum</i>, <i>F. mortiferum</i>, <i>F. ulcerans</i>, <i>F. varium</i> and <i>F. necrophorum</i>) and four subspecies of <i>F. nucleatum</i> (<i>nucleatum</i>, <i>animalis</i>, <i>vincentii</i> and <i>polymorphum</i>). By employing the PCR method, we investigated the prevalent features of <i>Fusobacterium</i> communities in Southern Chinese population and cancer patients. Surprisingly, we found <i>F. nucleatum</i> was dominant in both Southern Chinese population and cancer patients, and discovered the correlations of <i>Fusobacterium</i> species to host conditions. Moreover, <i>F. mortiferum</i> exhibited better diagnostic performance for cancers compared to other species, and the combination of <i>F. mortiferum</i>, <i>F. nucleatum</i>, body mass index and haemoglobin by a logistic regression model showed excellent diagnostic performances for cancers. Additionally, we determined the compositional features and loads of <i>Fusobacterium</i> communities in paired tumour, adjacent tissues and normal tissues of colorectal cancer and lung cancer. Hence, we developed a high-precision multiplex PCR assay to profile <i>Fusobacterium</i> in human faeces and tumour, and demonstrate its prevalence in Southern Chinese population with correlations to host conditions and cancers.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 12","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12740677/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145831703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering an AlkS-PalkB Transcription Factor-Based Biosensor With Improved Sensitivity to Isobutanol and Other Short-Chain Alcohols","authors":"Jiaming Chen,&nbsp;Xizi Wang,&nbsp;Can Zhu,&nbsp;Stefanie Frank,&nbsp;Jian Hao,&nbsp;Frank Baganz","doi":"10.1111/1751-7915.70288","DOIUrl":"10.1111/1751-7915.70288","url":null,"abstract":"<p>Isobutanol, a promising biofuel with higher energy content than ethanol, presents a sustainable alternative through biosynthesis. However, enhancing yield remains challenging due to the inefficiencies in microbial synthesis. This study introduces a transcription factor-based biosensor using the AlkS-PalkB system in <i>Escherichia coli</i>, which correlates green fluorescence with isobutanol concentration. Employing directed evolution, we modified AlkS to detect isobutanol, significantly improving biosensor specificity. Initial modifications increased the dynamic response from non-detectable to a 2.60-fold change. Subsequent optimisations through site-directed mutagenesis and promoter engineering further enhanced this response to a 5.56-fold change, equivalent to a 114% increase. Although engineered for isobutanol detection with high sensitivity, the engineered biosensor retains responsiveness to several short-chain alcohols. This biosensor provides a foundation for high-throughput screening of isobutanol and other short-chain alcohol-producing strains, though additional improvements in selectivity and operating range may be required for efficient implementation.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 12","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12740625/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145831731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing Salmonella Inhibition in Black Soldier Fly Larvae (Hermetia illucens L.) Conversion by Bioaugmentation With Gut Microbiota","authors":"Yuanpu Zhang,&nbsp;Wen Chen,&nbsp;Binbin Wang,&nbsp;Kashif ur Rehman,&nbsp;Arnold van Huis,&nbsp;Ahmed R. Henawy,&nbsp;Minmin Cai,&nbsp;Longyu Zheng,&nbsp;Zhuqing Ren,&nbsp;Feng Huang,&nbsp;Jibin Zhang","doi":"10.1111/1751-7915.70242","DOIUrl":"10.1111/1751-7915.70242","url":null,"abstract":"<p>Black soldier fly larvae (BSFL) can efficiently convert organic waste into biomass and reduce pathogenic bacteria in organic waste. The microbial composition of the substrate and the gut of BSFL is a pivotal factor in determining the efficacy of BSFL in pathogen elimination. However, there are insufficient data on the gut microbiology of BSFL in relation to pathogen inhibition. To address this gap, we investigated the dynamics of <i>Salmonella</i> during the conversion of chicken manure by BSFL and examined the role of intestinal bacterial communities and core bacteria in reducing <i>Salmonella</i> levels. The results indicate that BSFL treatment can reduce the amount of <i>Salmonella</i> in chicken manure, with the gut microbiome of the BSFL playing a crucial role in this reduction. Combining metagenomic analysis with culturomics methods, we isolated 158 strains from the larval gut, in which seven gut bacteria belonging to the genus <i>Bacillus</i> can promote BSFL to reduce <i>Salmonella</i>. In reinoculation and validation experiments, the combination of BSFL and <i>Bacillus velezensis</i> A2 enhanced the elimination of <i>Salmonella</i> from chicken manure and larvae. This study provides insight into how BSFL can reduce pathogenic bacteria in chicken manure and suggests that pairing BSFL with functional microorganisms can improve the biosafety of organic waste conversion by BSFL.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 12","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12721473/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145809128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phages Shape the Transformation of Organic Matter During Composting","authors":"Yuanyuan Bao,&nbsp;Jan Dolfing,&nbsp;Ruirui Chen,&nbsp;Chongwen Qiu,&nbsp;Jianwei Zhang,&nbsp;Xin Zhou,&nbsp;Liang Liu,&nbsp;Yiming Wang,&nbsp;Xiangui Lin,&nbsp;Youzhi Feng","doi":"10.1111/1751-7915.70291","DOIUrl":"10.1111/1751-7915.70291","url":null,"abstract":"<p>Microorganisms drive the biotransformation of dissolved organic matter (DOM) during organic wastes composting, yet the role of phages with different lifestyles (i.e., temperate and virulent) in this process remains poorly understood. Here, bulk metagenomic sequencing combined with electrospray ionisation (ESI) Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) was used to investigate the dynamics of temperate and virulent phage communities, microbial functional traits represented by the growth yield (Y)–resource acquisition (A)–stress tolerance (S) life-history strategies (Y-A-S) framework, and molecular changes in DOM composition, as well as their potential linkages during the composting of a rice chaff and chicken manure mixture. Our results revealed that the ratio of temperate/virulent phage, microbial Y/A strategy, and microbial-/plant-derived DOM components exhibited highly consistent dynamic patterns, all peaking during mid-composting stage when temperatures are elevated and remaining low at the initial and final stages. Random forest analysis further identified the ratio of temperate/virulent phages and the microbial Y/A strategy as key predictors of the variance in microbial Y/A trade-offs and microbial−/plant-derived DOM components, accounting for 10% and 13% of the explained variance, respectively. Together, our results demonstrate that an increased prevalence of temperate phages promoted the microbial Y-strategy and the accumulation of microbial-derived DOM components, while a greater dominance of virulent phages favoured the A-strategy and plant-derived DOM enrichment. These findings offer new insights into the ecological role of phages in mediating material transformation during organic waste composting.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 12","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12719608/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145802769","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Developing High-Efficiency Electroporation Protocols for Hard-To-Transform Halomonas spp.","authors":"André A. B. Coimbra,&nbsp;Ian J. White,&nbsp;Leonardo Rios-Solis","doi":"10.1111/1751-7915.70285","DOIUrl":"10.1111/1751-7915.70285","url":null,"abstract":"<p><i>Halomonas</i> species have recently emerged as attractive candidates for next-generation industrial biotechnology, due to their ability to thrive under high-salt conditions, which allows for fermentation under open, unsterile conditions. However, their genetic manipulation has long been hindered by difficulties in genetic transformation. In this study, we report the development of a highly efficient electroporation protocol for <i>Halomonas elongata</i> DSM 2581. By optimising competent cell preparation and electroporation parameters, and using plasmid DNA purified from <i>dam</i>/<i>dcm</i> methylation-deficient <i>Escherichia coli</i>, we achieved a maximum transformation efficiency of 2.8 × 10⁸ ± 0.2 × 10⁸ CFU/μg DNA—the highest efficiency reported for any <i>Halomonas</i> species to date. Notably, we observed that growing cells in low-salt medium and harvesting them at late-stationary phase considerably improved electroporation efficiency. Moreover, we further demonstrated that the use of non-methylated plasmids helped evade the defence systems of <i>H. elongata</i> DSM 2581 that target foreign DNA. Importantly, the protocol proved transferable to other industrially relevant strains, achieving efficiencies of 5.3 × 10⁷ ± 0.1 × 10⁷ and 5.4 × 10⁶ ± 1.2 × 10⁶ CFU/μg DNA in <i>Halomonas boliviensis</i> LC1 and <i>Halomonas campaniensis</i> LS21, respectively. Altogether, this work establishes a robust, high-efficiency electroporation method for <i>Halomonas</i> spp., facilitating future genetic manipulation and strain engineering work, as well as encouraging further research into underexplored <i>Halomonas</i> species.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 12","pages":""},"PeriodicalIF":5.2,"publicationDate":"2025-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12719606/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145802739","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}