Molecular Biotechnology最新文献

{"title":"Construction of a Yeast Two-Hybrid Library for Lilium × formolongi and Screening for Proteins Interacting with LfSVP.","authors":"Yu-Sha Pan, Zhen Zhou, Jin-Long Qian, Hai-Xia Chen, Ji-Ren Chen, Yu-Fan Li","doi":"10.1007/s12033-026-01567-0","DOIUrl":"10.1007/s12033-026-01567-0","url":null,"abstract":"<p><p>The MADS-box transcription factor SHORT VEGETATIVE PHASE (SVP) plays a pivotal role in flowering regulation. In this study, we aimed to elucidate the protein interaction network of LfSVP in Lilium × formolongi. A high-quality yeast two-hybrid (Y2H) cDNA library was constructed and screened using LfSVP as bait. From an initial pool of 18 candidate interactors, six proteins were confirmed through pairwise Y2H assays. These include known floral regulators APETALA1 (AP1), JASMONATE ZIM-DOMAIN (JAZ), SEUSS (SEU), and E3 ubiquitin-protein ligase ARI8 (ARI8), as well as stress-associated factors WRKY5 and SENSITIVE TO PROTON RHIZOTOXICITY1 (STOP1). Notably, virus-induced gene silencing (VIGS) demonstrated that LfSVP cooperates with LfAP1 to regulate floral organ development. While these findings suggest potential roles for LfSVP in flower development pathway, further functional validation is required in planta to establish the biological relevance of most interactions identified here. This work provides a potentially foundational interaction network for LfSVP and highlights candidates for future functional studies in lily flowering regulation.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3294-3304"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147390528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Protein Nanocages as Versatile Vectors for Nucleic Acid Delivery: Main Systems and Their Loading Mechanisms.","authors":"Carlos Francisco Coffeen, Aquetzali Estefanía Hernández-Gutiérrez, Ismael Bustos-Jaimes","doi":"10.1007/s12033-026-01564-3","DOIUrl":"10.1007/s12033-026-01564-3","url":null,"abstract":"<p><p>The rapid advancement of nucleic acid-based technologies, such as mRNA vaccines and CRISPR-based gene editing, has transformed modern medicine. Yet, their full therapeutic potential remains constrained by the need for precise cargo loading and efficient delivery. This limitation has inspired the development of novel vector systems capable of both protecting nucleic acids and directing them to specific cellular targets. Protein nanocages have emerged as a highly versatile and programmable class of delivery vectors. Considerable research has focused on repurposing and engineering various platforms, including virus-like particles, encapsulins, and ferritins, to serve as robust nanocarriers. Using innovative strategies, researchers have developed sophisticated cargo-loading mechanisms that have evolved from simple electrostatic attraction to high-specificity, affinity-based systems that mimic viral packaging. At the same time, a deeper understanding of how nucleic acids interact with the nanocage interior is revealing some key structural factors that drive efficient packaging. In this review, we highlight the key advances in the design and application of protein nanocages for nucleic acid delivery. We analyze the primary systems and their distinct loading mechanisms, discuss current challenges, and explore future directions to help establish engineered nanomaterials as a new and powerful category of medical therapies.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3091-3108"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13279676/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147372947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cloning and Characterization of a PL6 Alginate Lyase Aly94 from the Marine Bacteria.","authors":"Chunni Wu, Bohan Zeng, Zeting Ning, Wenwen Wang, Yuanpeng Wang, Qingdong Zhang, Danrong Lu","doi":"10.1007/s12033-026-01551-8","DOIUrl":"10.1007/s12033-026-01551-8","url":null,"abstract":"<p><p>Microbial alginate lyases are essential biocatalysts for analyzing alginate structure and sustainably producing bioactive alginate oligosaccharides (AOS). In this study, we characterized Aly94, a novel alginate lyase from the polysaccharide lyase family 6 (PL6) family, identified from a marine sediment metagenomic library. Biochemical analyses showed Aly94 exhibits optimal activity at 40 ℃ in 50 mM NaH₂PO₄-Na₂HPO₄ buffer (pH 7.0). Adding 20 mM NaCl significantly increases its catalytic efficiency. The enzyme exhibits a strong preference for polyguluronate (polyG) over polymannuronate (polyM), with specific activities of 4.19 U/mg (polyG), 0.25 U/mg (polyM), and 2.45 U/mg (alginate). When degrading substrates-particularly polyG-Aly94 primarily generates trisaccharides. Although Aly94 acts as an endolytic alginate lyase, it also could digest the monosaccharides from small oligosaccharide chains (∆G3, ∆G4). These catalytic properties, combined with its polyG-specific depolymerization, made Aly94 a promising candidate for biotechnological applications requiring controlled alginate saccharification and high-value AOS production.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3158-3170"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145959887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of Oncoviruses in Cancer Progression and Emerging Phytochemical-Based Therapies from Medicinal Plants.","authors":"Shivani Singh, Pardeep Yadav","doi":"10.1007/s12033-026-01561-6","DOIUrl":"10.1007/s12033-026-01561-6","url":null,"abstract":"<p><p>Oncoviruses continues to be an unattended cause of the global cancer load as they cause about 12-15 % of human malignancies in the world. The oncogenic infections that become persistent, such as Human Papillomavirus (HPV), Epstein-Barr Virus (EBV), Hepatitis B and C viruses (HBV, HCV), Human T cell Leukemia Virus-1 (HTLV-1), and Kaposi Sarcoma-associated Herpesvirus (KSHV), are associated with malignant transformation by continuous destabilization of host cell cycle regulation, immune surveillance, and metabolic homeostasis. These viruses abuse critical oncogenic signaling networks like PI3K/AKT/mTOR, NF -KB, JAK/STAT, MAPK, Wnt/β-catenin, and p53-dependent networks, hence, facilitating unchecked proliferation, chronic inflammation, genomic instability, and tumor progression. Although there have been improvements in the traditional forms of treatment, such as chemotherapy, radiotherapy, antiviral agents, immunotherapy, and gene-based treatments, yet clinical outcomes are still hampered by drug resistance, viral latency, systemic toxicity, and inaccessibility, and other severe side effects especially in low- and middle-income nations. Hence, there is a dire need to introduce new medicinal plant-based therapeutic approaches against oncovirus. This review analyses the molecular pathways of viral oncogenesis critically and discusses the clinical translation and a promising future potential of phytochemicals derived from medicinal plants like Phyllanthus emblica, Datura stramonium, Cannabis sativa, Andrographis paniculata, Aegle marmelos, Calotropis procera, and Prosopis cineraria proven to have bioactive compounds functioning as antivirals, immune-modulator, pro-apoptotic, and cell cycle regulatory effects in the preclinical models along with multi-targets.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3032-3050"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147458639","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An Integrated Computational and Biophysical Approach for Investigating the Structure-Function Impact of blaOXA-58 Mutations in Acinetobacter baumannii.","authors":"Huma Jalil, Hina Jalil, Shazia Shaheen Mir, Saad Amin, Kalsoom Khan, Asad Ullah, Ibrar Khan, Sadiq Azam, Sajjad Ahmad","doi":"10.1007/s12033-026-01553-6","DOIUrl":"10.1007/s12033-026-01553-6","url":null,"abstract":"<p><p>Carbapenems are the last-resort antibiotic option against A. baumannii infections, and Carbapenem resistance leads to the emergence of CRAB strains, which are difficult to treat. The CRAB novel reported mutation in the blaOXA-58, whose resistance mechanism was not explored. Here, we conducted molecular docking and molecular dynamics simulation to unveil and compare the molecular mechanisms of blaOXA-58 wild and mutant types (I87M, A88L, and double-mutated). The crystal structure of blaOXA-58 was retrieved from PDB, mutations were induced using the AlphaFold online server, and the structures of Carbapenem drugs (DOR, ETP, MEM, and IPM) were retrieved from PubChem database. Using PyRx 0.8, molecular docking was performed between Carbapenem drugs and protein blaOXA-58, both in wild-type and mutant variants. The results were validated through molecular dynamics simulations lasting 100 ns. Docking results showed the binding score of Carbapenem drugs with wild-type blaOXA-58 protein, I87M, A88L, and double-mutated forms, which revealed that the single mutation enhanced the binding affinity of Carbapenems toward the protein. Meanwhile, the decreased binding affinity of Carbapenem toward the double-mutated protein may be due to combined effect of the mutations on the protein structure. Overall, Ertapenem showed good binding scores (-8.1, -8.8, -8.8, and -6.1 kcal/mol), and Imipenem showed weak affinity (-6.3, -5.6, -6.3, and -5 kcal/mol) toward wild-type protein, I87M, A88L, and the double-mutated blaOXA-58 protein, respectively. Wild and double-mutated complexes were subjected to molecular dynamics simulation which revealed mutant-ETP was the most stable complex with low RMSD, RMSF, RoG, and B-factor values. PCA showed reduced flexibility, mutation affected surface loop arrangements in mutant complexes, raising SASA in DOR/IPM, reducing it in ETP, minimal effect in MEM. In contrast, mutant-IPM had higher RMSD, RMSF, RoG, lower hydrogen bonding, and better flexibility. Mutant-DOR exhibited a loss of α-helices and β-strands. The outcomes of the current analysis demonstrated that slight mutation in blaOXA-58 affects working of carbapenem antibiotic. The complicated impact of the double mutation on stability and binding should improve antibiotic-resistant pathogen research by enhancing targeted medication or inhibitor design.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3171-3191"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146030325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In-silico Analysis of the Cytochrome B Gene and its Role in Species Traceability from Fresh Meat Samples Through a Non-enzymatic DNA Extraction Method.","authors":"Muhammad Faisal, Rajwali Khan, Sarzamin Khan, Syed Muhammad Suhail, Ijaz Ahmad, Farhan Anwer Khan, Ahmad Faraz, Kamran Farid, Sania Qureshi, Amel Ayari-Akari","doi":"10.1007/s12033-026-01565-2","DOIUrl":"10.1007/s12033-026-01565-2","url":null,"abstract":"<p><p>In the present study, we first performed in-silico analysis and confirmed that the mitochondrial cytochrome b gene can be used for species differentiation, as it contains both conserved and variable regions, making it suitable for species identification. Furthermore, we used the non-enzymatic DNA extraction method as a novel procedure for DNA extraction from fresh meat samples. The results revealed that 6% of the collected samples were mislabeled, in which cattle meat was substituted with buffalo meat. The Chamkani tehsil of the target area showed the highest 12% mislabeling, followed by Shah Alam tehsil at 8% and Peshawar city tehsil at 4% in the collected samples. In duplex PCR, DNA fragments of two species (Goat/Sheep, Cattle/Buffalo) were amplified, and in multiplex PCR, DNA fragments of three species (Buffalo, cattle, and horse) were amplified for the target fragments of the cytochrome B gene. The present study concluded that the non-enzymatic DNA extraction method is equally effective for meat samples. Furthermore, we also found that peri-urban areas are prone to meat mislabeling, which needs vigilance from the concerned regulatory authorities.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3271-3280"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147481048","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plant-Based Oral Vaccines: Molecular Biotechnology Approaches Toward Functional Food-Based Immunization.","authors":"Lovepreet Singh, Harshita Jain, Shivani Jaswal, Richa Patel, Purusottam Banjare","doi":"10.1007/s12033-025-01550-1","DOIUrl":"10.1007/s12033-025-01550-1","url":null,"abstract":"<p><p>Plant-based oral vaccines are now becoming a more transformative medical form in immunotherapy; they are cheap and needle-free. These systems use natural bioencapsulation in plant cell walls, which preserves antigen stability within the gastrointestinal tract and promotes targeted release to mucosal immune tissues. Tests indicate that plant-derived antigens preserved their structure after exposure and they produced robust mucosal IgA and systemic IgG responses. From the early trial, such as seroconversion among human subjects consuming hepatitis B surface antigen (HBsAg)-expressing potatoes and high mucosal immunity generated by CTB-producing plants, promising results show their immunogenic potential. These technologies including chloroplast transformation that can produce more than 100 times higher antigen yields, CRISPR/Cas9 precision editing, and AI-equipped antigen design, further strengthen this platform. Although their relative strengths in standardization and poor digestibility are not widely available, plant-based oral vaccines have proven clear success with significant potential for immunization through functional foods, especially for resource-limited areas.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"2953-2971"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145912482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CRISPR-Cas9: Genome Engineering and Future Vaccine Applications.","authors":"Masoumeh Madhi, Pourya Gholizadeh","doi":"10.1007/s12033-026-01563-4","DOIUrl":"10.1007/s12033-026-01563-4","url":null,"abstract":"<p><p>The CRISPR-Cas9 system, a transformative genome engineering tool derived from prokaryotic adaptive immunity, is reshaping the landscape of biological research and therapeutic development. This review provides a critical synthesis of its rapidly evolving, yet underexplored, application in rational vaccine design. We analyze how CRISPR-Cas9 and its derivative platforms (including base editing, prime editing, and CRISPRi/a) are being repurposed from therapeutic gene editing to become indispensable assets in vaccinology. This transition is powered by the convergence of CRISPR-mediated precision with synthetic biology, enabling the rapid engineering of novel vaccine vectors and attenuated strains, the precise optimization of antigen sequences for enhanced breadth and potency, and the direct modulation of host immune responses. Notwithstanding this potential, significant technical and translational hurdles persist, including off-target editing risks, delivery inefficiencies in vivo, and unresolved regulatory pathways for genetically modified vaccines. We detail these mechanisms and evaluate the current preclinical and clinical landscape, while addressing persistent challenges in safety, delivery, and scalability. By delineating these advances and obstacles, this review outlines a forward-looking framework for leveraging CRISPR technology to create programmable, precision vaccines against emerging and re-emerging pathogens, moving the field beyond empirical methods toward a new paradigm of rational immunization.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3068-3090"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147468583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular Characterization of OsHAD Phosphoprotein: Expression, Catalytic Properties, and Functional Annotation.","authors":"Anuradha Pandey, Aparna Ramakrishnan, Raja Bhaiyya, Ravindra Pal Singh, Dipak Gayen","doi":"10.1007/s12033-026-01566-1","DOIUrl":"10.1007/s12033-026-01566-1","url":null,"abstract":"<p><p>Phosphoproteins and phosphatases play crucial roles in regulating diverse cellular processes in plants, including signal transduction, stress responses, and metabolic control, primarily through reversible phosphorylation. However, the functional and biochemical information on many rice phosphatases remains limited. In this study, we report the functional and biochemical characterization of OsHAD-2, a previously uncharacterized haloacid dehalogenase (HAD) family phosphatase from Oryza sativa.OsHAD-2 was heterologously expressed in Escherichia coli BL21 (DE3) and purified to homogeneity using GST-affinity chromatography. Enzymatic activity was assessed using p-nitrophenyl phosphate (pNPP), which confirmed that OsHAD-2 is a catalytically active phosphatase, with defined optimal reaction conditions, specific activity, and catalytic efficiency. Physical characterization of the purified protein was performed using circular dichroism (CD) spectroscopy, which revealed well-defined and stable secondary structure. In addition, mass spectrometry (MS) analysis validated the identity and purity of the recombinant protein. Further computational analysis provided insights into its structural features, stability determinants, and potential post-translational modifications. Together, these findings establish OsHAD-2 as a functional phosphatase and provide new biochemical and structural insights into the HAD-family phosphatase family, thereby advancing our understanding of rice phosphatase biology and laying a foundation for future studies on its role in plant physiology and stress adaptation.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3281-3293"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147343395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative Microbiome Analysis of Rhodiola fastigiata Rhizosphere Versus Bulk Soil in Xizang with Targeted Isolation of Rhizosphere-Derived Functional Strains.","authors":"Shuxin Zhou, Yao Yao, Ruiying Yuan, Weihua Chu","doi":"10.1007/s12033-026-01560-7","DOIUrl":"10.1007/s12033-026-01560-7","url":null,"abstract":"<p><p>Rhodiola fastigiata, a critically endangered medicinal plant of the Qinghai-Xizang Plateau, faces severe threats from habitat degradation. This study aimed to support its microbial-assisted conservation by characterizing the rhizosphere microbiome and isolating functional plant growth-promoting (PGP) bacteria. Using high-throughput sequencing of the 16S rRNA gene and ITS region, we found the rhizosphere community to be distinct and enriched in key taxa (e.g., Sphingomonas, Mortierella). Metabolic predictions suggested upregulated stress-adaptive pathways. Crucially, from 126 isolates, we obtained four Bacillus strains that concurrently produce protease, amylase, and cellulase and solubilize phosphate-quantifying a multifunctional PGP profile critical for nutrient-poor soils. These culturable, spore-forming strains provide direct resources for developing synthetic inoculants. Our work bridges microbial ecology with applied biotechnology, delivering both a foundational microbial map and candidate strains to enable the cultivation and conservation of this endangered species in extreme environments.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3259-3270"},"PeriodicalIF":3.2,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147284377","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}