BMC Biotechnology最新文献

{"title":"3D-printed metamaterial femoral prostheses via scalar field fusion and directional porous structure regulation.","authors":"Guoqing Zhang, Xiaoyu Zhou, Junxin Li, Yongsheng Zhou, Juanjuan Xie, Aibing Huang, Yuchao Bai","doi":"10.1186/s12896-026-01186-2","DOIUrl":"10.1186/s12896-026-01186-2","url":null,"abstract":"<p><strong>Background: </strong>This study integrates topology optimization, scalar field fusion (a method to realize smooth transition between solid and porous regions through implicit function interpolation), and directional porous structure control to develop a design method for customized metamaterial femoral prostheses, addressing key problems of conventional prostheses including poor implant compatibility, excessive bone resection, and insufficient long-term stability.</p><p><strong>Results: </strong>We initially completed the personalized design of femoral prostheses using a combination of retrograde and antegrade methods. This was followed by integrating topology optimization with porous structure filling to design the metamaterial. The prostheses were directly manufactured using 3D printing technology, and their performance was thoroughly evaluated. The results showed that the femoral prosthesis experienced the highest stress at 70° knee flexion (half-squat position), with a maximum stress of 1.269 × ⁴ MPa and a maximum displacement of 2.77 mm. Following topology optimization, although stress concentration and displacement slightly increased, the stress distribution became more uniform, resulting in a weight reduction of approximately 38.19%. Prostheses treated through scalar field fusion and adjustment of the filling direction of the porous structure exhibited metamaterial properties. The 3D-printed femoral prostheses and their matching components maintained high surface quality with no significant warping or deformation. Moreover, the porous structure displayed a well-defined pore configuration and connectivity. The prostheses and their components achieved a close fit, with dimensional accuracy and compatibility that generally met the requirements.</p><p><strong>Conclusions: </strong>After post-processing, the developed prototype serves as a valid proof-of-concept, thus establishing a foundation for the subsequent development and practical application of high-performance personalized biofix femoral prostheses.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13523339/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148275919","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":"Characterization of novel cellobiose 2-epimerases from Teredinibacter Haidensis and Cellvibrio Japonicus.","authors":"Yaxian Liu, Sabine Lutz-Wahl, Lutz Fischer","doi":"10.1186/s12896-026-01184-4","DOIUrl":"10.1186/s12896-026-01184-4","url":null,"abstract":"<p><strong>Background: </strong>Cellobiose 2-epimerases (CEs) are promising enzymes that catalyze the conversion of lactose to the bioactive disaccharides epilactose and lactulose. Low-temperature (≤8 °C) lactose bioconversion is essential in dairy applications to prevent microbial contamination. Unfortunately, most CEs are not particularly active at these low temperatures. Therefore, this work aims to discover and characterize novel cold-active CEs.</p><p><strong>Results: </strong>Two novel mesophilic CEs from Teredinibacter haidensis (ThCE) and Cellvibrio japonicus (CjCE) were characterized, which showed both epi- and isomerization activity additionally at low temperatures. ThCE showed a maximum epimerization activity (595.4 ± 4.4 nkat/mg protein) at pH 7.5 and 35 °C, while CjCE achieved maximum epimerization activity (887.4 ± 3.9 nkat/mg protein) at pH 9.5 and 40 °C. In addition, CjCE maintained an >80% activity across a wide pH (6-9.5) and temperature range (30-45 °C). Both CEs exhibited significant activity and stability at 8 °C. Specifically, ThCE and CjCE retained around 20 and 40% of their epimerization activity at 8 °C and showed half-lives of 96.3 and 72.2 days, respectively. Moreover, CjCE was also found to be capable of catalyzing the isomerization reaction at 8 °C. The isomerization activity of ThCE and CjCE increased ~370-fold (70.8 ± 2.7 nkat/mg protein) and ~230-fold (66.1 ± 0.3 nkat/mg protein), respectively, at lactose concentrations up to 600 mM (35 °C for ThCE and 40 °C for CjCE). This demonstrates how important it is to conduct investigations of CEs for lactose isomerization to lactulose up to the solubility limit of lactose.</p><p><strong>Conclusions: </strong>This study identified two new CEs that exhibit interesting catalytic properties for possible application in the generation of disaccharides with prebiotic properties from lactose at low temperatures.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"26 1","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13281645/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148275909","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 novel Carica papaya fruit peel peroxidase: kinetics, stability, and dye degradation.","authors":"Ozoemena E Eje, Rachael C Chinweuba, Okpanachi N Oyibo, Immaculata A Eze, Arinze L Ezugwu, Ferdinand C Chilaka","doi":"10.1186/s12896-026-01178-2","DOIUrl":"10.1186/s12896-026-01178-2","url":null,"abstract":"<p><p>Dyes in industrial effluents constitute a major source of environmental pollution, posing a threat to human life. The use of enzymes for bioremediation is a greener, more efficient approach than conventional methods. This study reported partial purification of a novel peroxidase (POD) from Carica papaya (pawpaw) peels using a two-step approach: 80% ammonium sulfate precipitation followed by gel filtration chromatography on Sephadex G-200. The enzyme was purified up to 24-fold with 23% yield and a specific activity of 6020.9 U/mg proteins. The molecular weight of the enzyme, determined by gel filtration, was 54.82 kDa, and the pH and temperature optima were 5.5 and 40 °C, respectively. The apparent Michaelis-Menten constants (K_m) for hydrogen peroxide (H₂O₂) and o-dianisidine were 0.026 and 0.93 mM, respectively. Metal ions: Co³⁺, K⁺, Ca²⁺, Na⁺, Fe²⁺, and Mn²⁺ inhibited activity, while Cu²⁺ was an activator in a concentration-dependent manner; 20 mM Zn²⁺ activated the enzyme. The enzyme retained 98.2% of its original activity after incubation at pH 7 and 40 °C for 30 min, while 49.8% was retained after 90 min. Peroxidase decolorized malachite green (up to 88.09%), eriochrome black T, local, methyl orange, methylene blue, and geni dyes. Hence, Carica papaya fruit peel peroxidase exhibited good stability after heat treatment. Meanwhile, the pronounced dye-decolorizing ability and the sustainable source of this peroxidase make it robust and cost-effective for biotechnological applications.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13508467/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148270191","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":"Green-synthesized quantum dots from quercus brantii for infected polymicrobial wound healing: mechanisms and biocompatibility.","authors":"Yasaman Sadat Nabipour, Ardeshir Hesampour, Salman Ahmady Asbchin, Maryam Tajabadi, Arman Rostamzad","doi":"10.1186/s12896-026-01183-5","DOIUrl":"10.1186/s12896-026-01183-5","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;The rise of antimicrobial resistance (AMR) has created an urgent need for alternative therapies, particularly against biofilm-driven polymicrobial wound infections. Although green-synthesized nanoparticles have emerged as promising candidates, many studies lack rigorous validation of their quantum properties, fail to clarify mechanisms of action, and omit essential cytotoxicity profiling in human cells.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Objective: &lt;/strong&gt;To synthesize silver (Ag), copper (Cu), and zinc oxide (ZnO) quantum dots (QDs) using Quercus brantii acorn extract, confirm quantum confinement effects, and evaluate their antimicrobial efficacy, mechanistic basis, and biosafety in a clinically relevant murine wound model.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;QDs were synthesized via a standardized hydrothermal approach. Quantum confinement was verified using photoluminescence (PL) spectroscopy and Tauc plot analysis. Antimicrobial activity was tested against multidrug-resistant (MDR) clinical isolates of Pseudomonasaeruginosa, methicillin-resistant Staphylococcus aureus (MRSA), Acinetobacter baumannii, and Klebsiella pneumoniae. To differentiate oxidative stress from chemical artifacts, we employed non-thiol reactive oxygen species (ROS) scavengers (Trolox, Mannitol) and electron paramagnetic resonance (EPR). Cytotoxicity was assessed in human keratinocytes (HaCaT) and dermal fibroblasts (HDF). In vivo efficacy was evaluated in a murine excisional wound model infected with a polymicrobial consortium, tracking pathogen-specific clearance. Hydrogel rheology and stability were also characterized.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;Monodispersed, crystalline QDs with evidence consistent with quantum confinement were obtained (Ag-QDs: 7.2 ± 1.5 nm; apparent bandgap: 2.85 eV). Ag-QDs showed strong antimicrobial activity (MIC: 4.5-18.1 µg/mL) and biofilm inhibition (up to 85% at ½×MIC). Mechanistic experiments, including EPR and ROS-scavenger assays, supported ROS-associated oxidative stress as a major contributor to bacterial killing under the tested conditions. Ag-QDs exhibited bactericidal activity at concentrations that were non-toxic to human skin cells in the employed assay (Selectivity Index for MRSA: 18.9). In vivo, topical application of an Ag-QD hydrogel (shear-thinning) accelerated wound closure to 95.3% by Day 14 (comparable to uninfected controls) and reduced the total bacterial burden by 4.2 log₁₀ CFU/g; all four pathogens were below the detection limit at the study endpoint. No overt systemic toxicity was observed based on the assessed biomarkers. Resistance development in serial passage assays was limited (2-fold MIC increase after 30 passages), noting that longer-term polymicrobial studies are required to fully evaluate resistance evolution.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;Q. brantii-mediated Ag-QDs are a promising green-synthesized nanomaterial with physicochemical characteristics consistent with quantum-dot behavior and not","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13501690/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148263165","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":"Library-based, multiplexed strategy for mapping protein interaction networks via crosslinking.","authors":"Yuxin Lu, Shuting Meng, Yi Zhang, Fei Wang, Tingting Shi, Yijia Chi, Song Chen, Dongxin Zhao","doi":"10.1186/s12896-026-01181-7","DOIUrl":"10.1186/s12896-026-01181-7","url":null,"abstract":"<p><strong>Background: </strong>Protein-protein interactions are fundamental to cellular function, yet resolving their interaction interfaces and dynamic behaviors in native biological contexts remains challenging, particularly for weak or transient interactions. Crosslinking strategies based on noncanonical amino acids offer an effective means to capture such interactions; however, traditional single-site incorporation provides limited coverage and may overlook critical interaction hotspots.</p><p><strong>Results: </strong>By employing a mutagenesis library, multiple interaction partners and cross-linking sites of a target protein can be simultaneously screened in a single experiment, without prior knowledge of its precise structural or functional features, enabling effective and unbiased analysis of its interaction network. In this study, we constructed an amber codon-scanning mutagenesis library of PSMD10, facilitating independent incorporation of the photocrosslinking ncAA p-azido-phenylalanine at multiple distinct residues. This approach allowed us to systematically interrogate and precisely map potential interaction regions across the protein surface. Coupled with crosslinking mass spectrometry, we identified multiple residues involved in intermolecular interactions, as well as previously unreported interaction partners, including T2FA, TBA1C, and ATRIP.</p><p><strong>Conclusions: </strong>These findings expand our understanding of PSMD10-associated proteasome interactome, demonstrate a multiplexed strategy for in situ mapping of protein interaction interfaces with broad coverage, and offer a valuable platform for developing therapeutics that target protein-protein interactions.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13551947/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148263195","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":"Au-IDE/LFA biosensor: an integrated sensing platform with lateral flow immunoassay for quantitative detection of hCG hormone using non-faradaic continuous FFT-admittance voltammetry.","authors":"Nazanin Behrooz, Farzaneh Shemirani, Reza Faridi-Majidi","doi":"10.1186/s12896-026-01175-5","DOIUrl":"10.1186/s12896-026-01175-5","url":null,"abstract":"<p><p>The development of the novel Au-IDE/LFA (gold interdigitated electrode with lateral flow immunoassay) biosensor, an integrated electrochemical sensing platform with lateral flow immunoassay, for quantitative detection of hCG (human Chorionic Gonadotropin) hormone levels is successfully directed here. To fabricate the Au-IDE/LFA setup, a free multi-electrode array of AU-IDE was configured exactly on five zones of LFA on the flow direction, employing non-faradaic measurements through a combination of two electrodes. The impedance behavior of each zone, connected with an Au surface electrode in the Au-IDE/LFA setup was characterized by frequency response analysis (FRA) using non-faradaic electrochemical impedance spectroscopy. Precisely, FRA validated that Au surfaces exhibit similar current homogeneity due to produce the repeatable impedance. The assay optimization using CFFT (Continuous Fast Fourier Transform)-admittance voltammetry, a new method, pinpointed that optimizing the excitation potential by utilizing lower SW frequencies at 88 Hz, amplitude at 10 mV, and DC potential at 10 mV resulted in excellent admittance responses of Au-IDE/LFA biosensor. Both bare and standard LFA were used for signal baseline establishment to ignore the background signal noise induced by the anti-β-hCG-AuNPs (6.01 µA/V), PBS buffer (11.51 µA/V), and urine background (22.11 µA/V). Admittance response showed a high sensitivity about three times more than impedance response with limit of detection by 2.53 mIU/mL in a linear range of 5-500 mIU/mL for hCG with R²= 0.998. This performance metric was utilized to determine the performance efficiency of proposed biosensor. The lack of any cross-interference with other human hormone like hCG and discriminating beta unit from alpha unit of hCG confirmed high selectivity and specificity of fabricated biosensor. The functional performance of biosensor resulted in the direct and quantitative detection of the native hCG in urine biological matrix, which offers a promising approach with high potential for early diagnosis of pregnancy test in clinical samples.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13491876/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148249065","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":"Rational design and high-yield expression of SARS-CoV-2 nucleocapsid antigen for a sensitive and specific in-house ELISA.","authors":"Abbas Tazarghi, Mehdi Behrouzikhah, Hamidreza Abbasi, Mohsen Ebrahimi, Alijan Tabarraei, Ahad Yamchi, Hadi Razavi Nikoo","doi":"10.1186/s12896-026-01177-3","DOIUrl":"10.1186/s12896-026-01177-3","url":null,"abstract":"<p><strong>Background: </strong>Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a global pathogen with a high prevalence rate. Among diagnostic methods, serological assays have been effective for rapid screening and identifying a history of infection or vaccination. The nucleocapsid protein is the most conserved and immunogenic antigen of SARS-CoV-2, making it a key target for such tests. Although commercial kits are available for this purpose, a custom design with superior specificity, sensitivity, and accuracy can be both clinically and economically advantageous. In the present study, we designed a recombinant nucleocapsid protein (rNP) using a multi-parameter suite of bioinformatic approaches, including codon optimization, ribosomal binding site engineering, mRNA secondary structure analysis, and optimized expression of the final construct in an E. coli system. The purified rNP was then applied in an in-house ELISA to compare its overall diagnostic efficacy with that of a commercially available kit on a pool of 37 predetermined positive and 15 negative sera for SARS-CoV-2.</p><p><strong>Results: </strong>Results showed that both assays were 100% specific, but the in-house design showed higher sensitivity (91.9% vs. 83.8%) and a greater area under the curve (0.94 vs. 0.84) compared to the commercial ELISA.</p><p><strong>Conclusion: </strong>This demonstrates that our optimized rNP production platform enables the development of a high-performance serological assay with superior diagnostic accuracy compared to a commercial alternative, providing a powerful and economical tool for surveillance and research in the post-pandemic era.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13491942/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148249114","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":"Enhanced anti Pseudomonas aeruginosa efficacy of niosomal-encapsulated Frondoside A from sea cucumber: a novel biotechnological approach.","authors":"Tohid Piri-Gharaghie, Maryam Hassanpoor, Neda Jegargoshe-Shirin, Mona Soosanirad, Shahoo Khayati, Ali Farhadi-Biregani","doi":"10.1186/s12896-026-01172-8","DOIUrl":"10.1186/s12896-026-01172-8","url":null,"abstract":"<p><strong>Background: </strong>Pseudomonas aeruginosa (P. aeruginosa) forms persistent biofilms that limit antibiotic efficacy. Frondoside A, a triterpenoid glycoside from sea cucumbers, possesses antimicrobial potential but suffers from poor solubility and stability. This study aimed to enhance its antibacterial and anti-biofilm activities via niosomal encapsulation and to assess its effect on biofilm-related gene expression.</p><p><strong>Methods: </strong>Niosomal Frondoside A was synthesized by thin-film hydration and characterized for size, charge, and encapsulation efficiency. Antibacterial activity was determined using agar diffusion and MIC/MBC assays, while biofilm inhibition was quantified by crystal violet assay. Expression of GacS and PslA was analyzed by qRT-PCR with 16 S rRNA as an internal control.</p><p><strong>Results: </strong>The optimized niosomes had a zeta potential of - 8.6 mV, and 78 ± 0.408% encapsulation efficiency. Compared to the free drug, the niosomal formulation reduced MIC/MBC values from 6.25/12.5 mg mL⁻¹ to 1.56/3.125 mg mL⁻¹, enlarged inhibition zones by 45-60%, and decreased biofilm biomass by 68-79% (p < 0.001). Furthermore, qRT-PCR showed significant downregulation of GacS and PslA genes.</p><p><strong>Conclusions: </strong>Niosomal encapsulation markedly improves the antimicrobial and gene-suppressive efficacy of Frondoside A, offering a potent nanocarrier approach against multidrug-resistant P. aeruginosa biofilms.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13459401/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148194371","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":"Electroporation-mediated site-specific knock-in of a human coding sequence into the bovine ALB locus in embryos.","authors":"Kyeong-Hyeon Eom, Soo-Young Yum, Gyeong-Min Gim, Woo Jae Son, Young-Chai Kim, Jae-Yoon Choi, Dong-Hyeok Kwon, Goo Jang","doi":"10.1186/s12896-026-01180-8","DOIUrl":"10.1186/s12896-026-01180-8","url":null,"abstract":"<p><strong>Background: </strong>Efficient delivery of genome-editing tools into embryos is essential for the production of gene-edited cattle. Somatic cell nuclear transfer and microinjection are commonly used techniques, but electroporation has recently gained attention owing to its technical simplicity, high-throughput capacity, and suitability for large-scale applications. However, its application has been largely restricted to gene knockout owing to the presence of the zona pellucida and size limitations of deliverable materials. To address these challenges, in this study, we optimized electroporation-mediated knock-in (KI) strategies in bovine embryos using single-stranded oligodeoxynucleotides (ssODNs) and adeno-associated virus (AAV) vectors as donors.</p><p><strong>Results: </strong>Initial experiments using ssODN donors with asymmetric homology arms (10 bp left and 25 bp right) resulted in a KI blastocyst (BL) frequency of 13.9 ± 14.1%, although outcomes varied widely across replicates. By contrast, AAV donors carrying 61 bp cargo flanked by 500 bp homology arms enabled consistent KI events, after optimizing the concentration of AAV and length of homology arm. Using the optimized system, a 2.2 kb human albumin coding sequence was successfully inserted into the bovine albumin locus, yielding KI BL frequency of 55.6 ± 41.6%.</p><p><strong>Conclusions: </strong>Collectively, these findings demonstrate that electroporation combined with AAV-mediated donor delivery represents an efficient strategy for generating transgenic bovine embryos, which is a crucial first step toward producing transgenic cattle as bioreactors for high-value protein production.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13459287/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148194415","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":"Unveiling a putative class 3 L-asparaginase in Glutamicibacter sp.: from soil isolation to in silico characterization.","authors":"Mahdis Mofidi, Gholamhossein Ebrahimipour, Maryam Azimzadeh Irani, Ali Etemadi, Mohammad Yaghoubi-Avini","doi":"10.1186/s12896-026-01179-1","DOIUrl":"10.1186/s12896-026-01179-1","url":null,"abstract":"<p><strong>Background: </strong>Type II L-asparaginases are important therapeutic agents for treating childhood leukemia, but numerous side effects of these enzymes, besides neutralization by antibodies due to repeated injection, prompt extensive research to identify alternatives in other classes of this enzyme.</p><p><strong>Results: </strong>Among the six asparaginase-producing strains, the best-producing isolate was identified as Glutamicibacter. The asparaginase and glutaminase activities in culture supernatant were 15 U/ml and 1.5 U/ml, respectively. The full-length asparaginase gene was sequenced, and the phylogenetic tree revealed strong clustering with recently identified class 3 asparaginases from Rhizobium etli. The presence of key residues from Class 3 ReAV and ReAIV further confirms this similarity. The AlphaFold 3 server confidently predicted the enzyme's 3D structure. Molecular docking of L-asparagine and L-glutamine using Rosetta revealed binding energies of -9.99 REU for both substrates that were higher than those of the Type IV and V enzymes. Molecular dynamics results indicated that enzyme-Asn was more stable than enzyme-Gln with average RMSD values of 0.33 nm and 0.44 nm, respectively. Enzyme-Asn reached a relatively stable plateau after 0.05 ns, whereas enzyme-Gln showed a sudden jump after 0.75 ns, indicating the separation of glutamine from the protein. A comparison of the antigenic determinants of GlAVI with those of ReAIV, ReAV, EcAII, and ErAII revealed that GlAVI exhibited lower immunogenicity than the other enzymes. Allergenic epitopes were predicted using bioinformatics, demonstrating that nonallergenic epitopes cover a significant portion of the enzyme.</p><p><strong>Conclusion: </strong>The newly isolated Glutamicibacter L-asparaginase appears to be a new type of class 3 asparaginases with high potential for novel therapeutic drug development because of its less and dissimilar immunological properties.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":" ","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13455196/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148161059","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}