Applied Microbiology and Biotechnology最新文献

{"title":"Knockout of glycerol metabolic pathways enables efficient mycolicibacterial phytosterol conversion using glycerol as cosovlent.","authors":"Shikui Song, Ye Wang, Jinpeng Yu, Qianqian Ma, Xiyao Cheng, Baoguo Zhang, Zhengding Su","doi":"10.1007/s00253-024-13360-7","DOIUrl":"https://doi.org/10.1007/s00253-024-13360-7","url":null,"abstract":"<p><p>Mycobacterial phytosterol conversion naturally occurs within a vegetable oil environment, a process that has been widely adopted in water‒oil two-phase fermentation to produce active pharmaceutical intermediates of steroids. The use of hydroxypropyl-β-CD (HP-β-CD) as a potential replacement for vegetable oils has been explored. However, both approaches encounter challenges, particularly the need for a high concentration of either vegetable oil or HP-β-CD in the fermentation medium, which significantly affects the efficiency of mycobacterial phytosterol conversion as well as the recovery of the resulting products. In this study, we demonstrated that the knockout of glycerol metabolic pathways enables the utilization of glycerol as cosovlent for efficient phytosterol conversion in Mycolicibacterium neoaurum HGMS6, an industrial strain known for its production of 4-androstene-3,17-dione (4-AD). Our bioinformatic analysis revealed two parallel glycerol metabolic pathways in the HGMS6 strain. These pathways are independently initiated by glycerol kinase (GLPK) and glycerol dehydrogenase (ADHC). To investigate their roles, we conducted gene knockout experiments in which both the glycerol kinase gene (glpk) and the glycerol dehydrogenase gene (adhc) were targeted in HGMS6, resulting in the creation of a glycerol metabolism-deficient mutant, denoted HGMS6^{Δadhc/Δglpk}. This double mutant exhibited complete inactivation of mycobacterial glycerol metabolism without any adverse effects on mycobacterial growth when cultured in conventional fermentation medium containing glucose. We then evaluated HGMS6^{Δadhc/Δglpk} in an aqueous phase fermentation medium and determined that fermentation media consisting of 10% glycerol and 0.5% HP-β-CD provided the optimal conversion efficiency. In pilot-scale fermentation experiments, this mutant was highly efficient at converting phytosterols, achieving a remarkable conversion rate of up to 83.8% (mol/mol%). This bioconversion level was on par with that of HGMS6 and HGMS6^{Δadhc/Δglpk} when operating in vegetable oil- or HP-β-CD-containing fermentation media. This study provides a promising strategy for enhancing the efficiency of mycobacterial phytosterol conversion. KEY POINTS: • TheGLPK and ADHC of M. neoaurum were confirmed and examined. • The enzymology of ADHC and GLPK was characterized in vitro. • An adhc/glpk-knockout mutant efficiently converts phytosterols in fermentation media containing 10% (w/v) glycerol and 0.5%(w/v) HP-β-CD.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"169"},"PeriodicalIF":3.9,"publicationDate":"2025-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144673813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a chemotactic SynCom bioorganic fertilizer for biocontrol of bacterial wilt in tobacco fields.","authors":"Yanxia Liu, Weiqi Zhang, Li Guo, Han Li, Jingwei Zhu, Xiang Li, Xiaoni Zhang, Xinrun Yang, Yangchun Xu, Qirong Shen, Tianjie Yang, Zhong Wei","doi":"10.1007/s00253-025-13547-6","DOIUrl":"10.1007/s00253-025-13547-6","url":null,"abstract":"<p><p>Bacterial wilt caused by Ralstonia solanacearum is one of the most severe plant diseases all over the world. Currently, many scientists are using SynCom (synthetic microbial community) to control this disease. However, designing of highly efficient SynCom remains challenging. In this study, we isolated 372 bacteria with different morphologies from the rhizosphere soil of healthy tobacco plants in a diseased field. Based on the antagonistic activity, compatibility, and presence of cheA gene, a marker gene of chemotaxis, we constructed a chemotactic SynCom comprising two Pseudomonas strains and one Bacillus strain. In vitro experiments revealed that vitamin C, propionic acid, and esculetin significantly promoted the growth and antagonistic ability of chemotactic SynCom strains. Next, we optimized the proportion of organic fertilizer to support the growth of the SynCom strains. The application of the bioorganic fertilizer containing chemotactic SynCom can protect diseased tobacco field from R. solanacearum invasion and enhance yields. Finally, rhizosphere bacterial community analysis showed that phyla Bacillota and Chloroflexiota were significantly enriched. This study highlights the potential of chemotactic SynCom for effective biocontrol, offering a new approach for managing bacterial wilt and enhancing crop yields. KEY POINTS: • The bioorganic fertilizer containing chemotactic SynCom suppresses R. solanacearum and boosts tobacco yields. • Root exudate-selected prebiotics enhance the antagonistic effects of SynCom against pathogens. • SynCom bioorganic fertilizer regulates the rhizosphere microbiome, enriching beneficial bacterial phyla.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"168"},"PeriodicalIF":3.9,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12267373/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144648404","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":"Multi-walled carbon nanotubes as reusable boosters of pyocyanin production for anticancer research.","authors":"Joanna Honselmann Genannt Humme, Kamila Dubrowska, Magdalena Perużyńska, Marek Droździk, Radosław Birger, Martyna Jurkiewicz, Tomasz Kędzierski, Ewa Mijowska, Tomasz Idzik, Jacek G Sośnicki, Elżbieta Filipek, Mateusz Piz, Rafał Rakoczy, Adrian Augustyniak","doi":"10.1007/s00253-025-13543-w","DOIUrl":"10.1007/s00253-025-13543-w","url":null,"abstract":"<p><p>Carbon nanotubes (CNTs) emerged as nanomaterials with a wide variety of applications, e.g., as boosters of bioprocesses efficiency. The stimulation of the production of the blue pigment called pyocyanin is one of numerous examples. Moreover, its importance comes from the potential anticancer properties of the pigment. Therefore, this contribution evaluated different commercially available multi-walled carbon nanotubes (MWCNTs) in pyocyanin production using the Design of Experiment methodology. The interactions between pigment-producing bacteria and nanomaterials were revealed as well. Moreover, the purified pigment was tested against normal and cancer cell lines. Interestingly, the results showed that all tested CNTs stimulated pyocyanin production. The most effective CNTs were used in the process optimisation in terms of temperature (32 °C) and carbon nanomaterial concentration (812 μg/mL). It was also revealed that the optical density and viability of the bacterial culture were elevated, while the pyoverdine production was decreased. Furthermore, no oxidative stress was detected. Moreover, the confocal microscopy study indicated that the cells surrounded the aggregates of MWCNT and produced more proteins within the biofilm structure, compared to the control experiment. The tests on neoplastic cell lines showed an excellent antiproliferative activity of pyocyanin against melanoma without pronounced adverse effects on normal fibroblasts. The nanomaterial incorporated in the bioprocess was successfully reused, making the method sustainable and cost-effective. KEY POINTS: • The stimulative effect of nanomaterial on pyocyanin production was optimised • Nanomaterial can be reused in the bioprocess without losing the stimulative effect • Pyocyanin exhibits significant antiproliferative action against melanoma.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"167"},"PeriodicalIF":3.9,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12259743/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144625219","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":"Genomically-selected antifungal Bacillaceae strains improve wheat yield and baking quality.","authors":"Alejo Casal, Fernán Oscar Gizzi, Sol Agostina Figueroa, Tomás Denis Petitti, Facundo Ferragutti, Jimena Gaido, Mariano Alberto Torres Manno, Gabriel Céccoli, Luciana Paoletti, Christopher Dunlap, Lucas Damián Daurelio, Martín Espariz","doi":"10.1007/s00253-025-13544-9","DOIUrl":"10.1007/s00253-025-13544-9","url":null,"abstract":"<p><p>Soil microbial diversity degradation through agricultural intensification necessitates sustainable alternatives. This study employed genomic and phenotypic approaches to characterize wheat rhizosphere-associated Bacillaceae for agricultural applications. Initial screening of 576 sporulating isolates for antifungal activity against Fusarium graminearum, followed by RAPD analysis, identified 39 distinct genetic profiles, out of which 15 were classified in Bacillus amyloliquefaciens or Priestia megaterium groups by 16S RNA sequence. Whole-genome sequencing of selected strains enabled precise taxonomic classification and comprehensive trait prediction using in silico tools. Genomic mining revealed strain-specific distributions of beneficial traits, including antimicrobial compound production pathways and plant growth-promoting characteristics. Phenotypic validation confirmed key predicted traits while uncovering additional functionalities not detected in silico. Integration of kernel bioassays, pot experiments, and field trials identified Bacillus velezensis ZAV-W70 and P. megaterium ZAV-W64 as promising biofertilizer and biocontrol candidates, demonstrating enhanced yield without fungicides and improved bread-making quality, respectively. These findings highlight the value of combining genomic analysis with traditional screening methods for developing effective agricultural biologicals, contributing to sustainable wheat production practices. KEY POINTS: • Rhizosphere Bacillaceae strains show dual plant growth promotion and biocontrol • B. velezensis ZAV-W70 and P. megaterium ZAV-W64 increase wheat yield • ZAV-W64 increases bread-making quality including total gluten and alveograph W.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"164"},"PeriodicalIF":3.9,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12241182/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144599246","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":"Application of MALDI-TOF MS and FT-IR spectroscopy in identification and antibiotic resistance profiling of lactic acid bacteria.","authors":"Tamás Kocsis, Anna Győrffy, Andrea Pomázi","doi":"10.1007/s00253-025-13550-x","DOIUrl":"10.1007/s00253-025-13550-x","url":null,"abstract":"<p><p>This study evaluates the combined application of MALDI-TOF MS and Fourier transform infrared (FT-IR) spectroscopy for the purpose of species identification and strain-level differentiation of lactic acid bacteria isolated from commercial yoghurts. MALDI-TOF MS provided rapid species-level identification, while FT-IR detected metabolic variations by analysing key cellular components such as membrane fatty acids (3000-2800 cm^-1), protein amide bands (1800-1500 cm^-1), and polysaccharides (1200-900 cm^-1). These spectral features reflected phenotypic differences among strains linked to antibiotic resistance profiles. Disc diffusion antibiotic susceptibility testing revealed significant variability among isolates, and a strong correlation was observed between FT-IR-derived linear discriminant analysis (LDA) clusters and resistance to oxacillin, clindamycin, and tetracycline (Chi² test, p < 0.05). This demonstrates FT-IR as a rapid, non-invasive tool for the early detection of resistant strains, facilitating real-time monitoring of bacterial adaptation during fermentation. The findings of this study provide support for integrating FT-IR and MALDI-TOF MS into industrial quality control, aiding in strain selection and enhancing food safety. KEY POINTS: • FT-IR enables rapid phenotypic typing of lactic acid bacteria strains. • Spectral profiles correlate with antibiotic resistance. • MALDI-TOF MS and FT-IR offer a robust, complementary identification method.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"165"},"PeriodicalIF":3.9,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12245945/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144599245","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":"Prediction model based on gut microbiota as a non-invasive tool for gastric cancer diagnosis.","authors":"Changchang Chen, Chen Chen, Xiaoguang Zheng, Weizhong Wang, Jian Shen, Gulei Jin, Jianxin Lyu, Lijun Lin","doi":"10.1007/s00253-025-13548-5","DOIUrl":"10.1007/s00253-025-13548-5","url":null,"abstract":"<p><p>Gastric cancer (GC) is a malignant cancer of the digestive tract with high morbidity and mortality. Previous studies have shown that current diagnostic methods largely rely on invasive procedures. Moreover, there are no highly sensitive and accurate biomarkers available for early GC diagnosis. Recent studies using 16S rRNA technology show that gut microbiota can differentiate between diseased and healthy individuals. However, fewer studies emphasize the gut microbiome's value in GC diagnosis. In this study, we collected 455 fecal samples, including 100 from healthy individuals (healthy controls [HCs]), 153 from GC patients, 43 from patients with non-neoplastic diseases of the stomach, and 159 from verification individuals. Our analysis revealed a significantly increased microbial richness in the GC group (Chao1 index, P < 0.05) and distinct compositional differences (principal coordinates analysis). Linear discriminant analysis effect size analysis identified 19 HC-enriched genera (e.g., Bacteroides) and 31 GC-enriched genera (e.g., Streptococcus). The random forest model selected 20 key diagnostic genera, achieving an area under the receiver operating characteristic curve (AUC) of 0.81. By integrating 10 tumor biomarkers, the combined diagnostic model improved the AUC to 0.86 (validation set: 0.84). Tumor biomarker positivity (60.78%) did not directly correlate with microbiota, but the microbiota-biomarker model improved non-invasive diagnostic accuracy, providing a new approach for early GC screening. KEY POINTS: • Changchang Chen and Chen Chen contributed equally to this work • Gut microbiota changes significantly in gastric cancer • Microbiome shows promise as non-invasive diagnostic markers • The combined microbiota-tumor marker model improves diagnosis.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"166"},"PeriodicalIF":3.9,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12245982/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144607257","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":"The engineering toolbox of Parageobacillus thermoglucosidasius.","authors":"Marie Millgaard, Belinda Escher, Ivan Pogrebnyakov, Alex Toftgaard Nielsen","doi":"10.1007/s00253-025-13508-z","DOIUrl":"10.1007/s00253-025-13508-z","url":null,"abstract":"<p><p>The thermophilic bacterium Parageobacillus thermoglucosidasius is considered a promising host for industrial microbial production. However, its genetic engineering toolbox is still under development. The focus of this review is to provide an organised summary of the currently available resources to facilitate the use of this organism. The article offers a comprehensive overview of regulatory parts used for the construction of genetic circuits and plasmids, including promoters, ribosome binding sites, terminators, antibiotic resistance genes, reporter genes, and other genetic elements of interest. Additionally, it examines the developments in P. thermoglucosidasius vector designs over the years. Here, vectors were categorised either as replicative vectors intended for gene expression, or integrative vectors intended for genomic engineering. The functionality of each vector was described, and their contributions to the progress of molecular tools available for P. thermoglucosidasius were evaluated. The review also summarises recent advancements in CRISPR-based systems relevant to this organism. Finally, this review discusses potential improvements that could further contribute to the expanding engineering toolbox of P. thermoglucosidasius, paving the way for more advanced applications. KEY POINTS: • The existing engineering toolbox for P. thermoglucosidasius contributes to the growing interest in using it as a thermophilic production host. • The engineering toolbox for P. thermoglucosidasius has expansion potential in genetic circuit parts and CRISPR-based tools. • Thermophilic hosts like P. thermoglucosidasius are in need of more thermostable reporter genes and thermostable selection markers.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"163"},"PeriodicalIF":3.9,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12241125/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590360","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":"Bacillus spore showcasing key antigens of African swine fever potentializes a promising oral vaccine candidate.","authors":"Xiaohua Wang, Baochao Fan, Tao Zhu, Hanwen Shen, Lian Li, Sihan Wang, Xiulian Yin, Baoxia Tian, Jinfeng Zhang, Bin Li, Chuping Luo","doi":"10.1007/s00253-025-13511-4","DOIUrl":"10.1007/s00253-025-13511-4","url":null,"abstract":"<p><p>African swine fever (ASF) remains a widespread and fatal infectious disease affecting both domestic pigs and wild boars around the world, and there is still no vaccine available for full protection. In recent years, Bacillus spores have been developed into effective vaccine carriers. Here, Bacillus amyloliquefaciens spores from strain CPLK1314 were used as a vaccine delivery system to display three highly antigenic proteins, p72, p30, and p54 of ASF, and their immune responses in BALB/c mice were evaluated. Our results demonstrated that the antibiotic-free Bacillus gene editing system constructed in this study can work effectively, and p72 and the fusion antigen p3054 were successfully expressed in the spore coat of CPLK1314 by fusing with CotB and CotG, respectively. Further animal experiments showed that the recombinant spores ΔBa72 and ΔBa3054 exhibited good immunogenicity, which induced high levels of antigen-specific serum IgG, proliferation of splenic lymphocytes, secretion of immune-related factors, as well as a significant increase in the proportion of T lymphocytes CD3⁺ CD4⁺ and CD3⁺ CD8⁺, and B lymphocytes B220⁺ CD19⁺. Interestingly, when providing the same number of spores, a 1:1 mixture of the two, ΔBa72 + 3054, could simultaneously trigger similar levels of both p72 and p30 antibodies without cross-affecting production levels. Therefore, based on the CRISPR-Cas9 continuous gene editing system constructed in this study, the simultaneous co-expression of one antigen species or synergistic expression of multiple antigens at different spatial positions of B. amyloliquefaciens spores might make it a promising candidate for an ASF oral vaccine. KEY POINTS: • Bacillus amyloliquefaciens spore is an ideal and effective vaccine delivery system • Bacillus spore showcasing key antigens of ASF is a promising oral vaccine candidate • The Bacillus vaccine can trigger effective protective responses in animals.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"162"},"PeriodicalIF":3.9,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12238207/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582934","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":"Prevalence and genomic insights into Yersinia enterocolitica in Southeastern China (2008-2022).","authors":"Lei Fang, Shuxuan Li, Jie Rong, Shengkai Li, Yuwen Zhang, Huihuang Lou, Zhongbi Xie, Yuqin Hu, Yuejin Wu, Airong Xie, Yi Li","doi":"10.1007/s00253-025-13500-7","DOIUrl":"10.1007/s00253-025-13500-7","url":null,"abstract":"<p><p>Yersinia enterocolitica is a significant foodborne pathogen causing gastrointestinal illnesses worldwide. This study investigates the prevalence and genomic characteristics of Y. enterocolitica to assess potential health risks in southeastern China, a region lacking mandatory yersiniosis monitoring. From 2939 samples collected between 2008 and 2022, 105 isolates were recovered. The highest prevalence was found in rodents (8.1%), followed by retail meats (7.1%), other foods (3.7%), and human clinical cases (0.8%). In addition to meats and rodents, ready-to-eat salads, seafood, and frozen food products were identified as potential transmission vehicles. Various bioserotypes and sequence types (STs) was identified, including twelve previously unreported STs. Biotype 1A, exhibiting greater genetic diversity than more pathogenic biotypes (3 and 4), was frequently found in human clinical cases. Phylogenetic analysis revealed two main lineages, with isolates primarily clustered by biotype and pathogenic traits. Antimicrobial susceptibility testing revealed 46.7% (49/105) of isolates were multidrug resistant (MDR), with frequent resistance to polymyxin B (100%), azithromycin (50.5%), and sulfanilamide isoxazole (31.4%). These findings highlight the ecological complexity and diversity of Y. enterocolitica, especially non-pathogenic biotype 1A strains, and underscore the need for enhanced food safety and antimicrobial stewardship to mitigate the public health impact of Y. enterocolitica infections. KEY POINTS: Biotype 1 A strains exhibited greater genetic diversity than pathogenic biotypes. Pathogenic strains were mainly associated with lineage HC1490_2, not HC1490_10. Higher MDR levels were observed in biotype 3 and 4 strains.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"161"},"PeriodicalIF":3.9,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12234613/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144574745","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":"Novel centromeric plasmid for stable extrachromosomal gene expression in Aurantiochytrium limacinum.","authors":"Person Pesona Renta, Cian-Huei Syu, Ta-Yu Huang, Yi-Ting Chang, Yu-Feng Liang, Ssu-Ting Chen, Po-Wei Weng, Ming-Chen Hsu, Keng-Hung Lin, Tsunglin Liu, Anna C-C Jang, Che-Chia Tsao, Han-Jia Lin, Hung-Yun Lin, Yi-Min Chen","doi":"10.1007/s00253-025-13527-w","DOIUrl":"10.1007/s00253-025-13527-w","url":null,"abstract":"<p><p>Aurantiochytrium limacinum holds great promise for producing sustainable single-cell oil as an alternative to fish oil. However, research into its complex biological and biochemical characteristics and efforts toward strain improvement have been hampered by insufficient genetic tools. Until now, genetic transformations of A. limacinum have relied solely on chromosome integration, which is inefficient and prone to insertional mutagenesis and other issues related to genetically modified organisms (GMOs). This paper describes the first centromeric plasmid for A. limacinum. Amplification of this shuttle vector by E. coli enables direct delivery into A. limacinum via electroporation, where it undergoes stable replication and segregation into daughter cells. The key to the stable plasmid maintenance lies in a 500 bp segment derived from chromosome 24 of Phaeodactylum tricornutum. While this segment does not significantly enhance the efficiency of vector transformation, it enables the replication and maintenance of the shuttle vector in the host cell as closed circular DNA. The plasmid from three transformants demonstrates a high segregation efficiency of 96.8 ± 0.3% (n = 3), even in the absence of antibiotic selection. This novel centromeric plasmid considerably enhances the flexibility of genetic manipulations and gene expression in A. limacinum, opening new avenues for its study and industrial application. KEY POINTS: • First centromeric plasmid developed for genetic transformation in A. limacinum. • The novel plasmid enhances flexibility in genetic manipulation and gene expression. • The plasmid achieves 96.8 ± 0.3% (n = 3) segregation efficiency without antibiotic selection.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":"160"},"PeriodicalIF":3.9,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12227449/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558908","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}