Molecular Biotechnology最新文献

{"title":"Thermal Effect on Algae, Biofilm and Their Composition Towards Membrane Distillation Unit: A Mini-review.","authors":"Yin Sim Ng, Derek Juinn Chieh Chan","doi":"10.1007/s12033-023-00853-5","DOIUrl":"10.1007/s12033-023-00853-5","url":null,"abstract":"<p><p>Membrane distillation (MD) has lower operating temperature and potential to recycle waste heat for desalination which catches much attention of the researchers in the recent years. However, the biofouling is still a challenging hurdle to be overcome for such applications. The microbial growth rate, secretion and biofilm formation are sensitive to heat. Membrane distillation is a thermally driven separation, so the increase of temperature in the seawater feed could influence the extent of biofouling on the unit parts. In this review, we present the effect of temperature on algal growth, the range of temperature the microbes, marine algae and planktons able to survive and the changes to those planktons once exceed the critical temperature. Thermal effect on the biofilm, its composition and properties are discussed as well, with association of the biofilm secreting microbes, but the study related to membrane distillation unit seems to be lacking and MD biofouling factors are not fully understood. Characterization of the algae, biofilm and EPS that govern biofouling are discussed. This information not only will help in designing future studies to fill up the knowledge gaps in biofouling of membrane distillation, but also to some extent, assist in pointing out possible fouling factors and predicting the degree of biofouling in the membrane distillation unit.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3512-3522"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10495708","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":"A CHO-Derived Matrix Attachment Region Enhances Transgene Dosage, SATB1 Recruitment, and Monoclonal Antibody Expression in a Dual-Promoter Vector System.","authors":"Sarmishta Majumdar, Rupashree Salvi, Prajakta Dandekar, Ratnesh Jain","doi":"10.1007/s12033-026-01577-y","DOIUrl":"10.1007/s12033-026-01577-y","url":null,"abstract":"<p><p>The production of monoclonal antibodies (mAbs) in Chinese hamster ovary (CHO) cells is often affected by position-effect variegation and the gradual loss of transgene expression over time. Hence, we have designed a dual-promoter IgG expression vector and compared versions that either contained or lacked a CHO-derived matrix-attachment region (MAR). Stable CHO-S pools, cultured in serum-free conditions, revealed that the MAR-containing construct produced higher and more consistent antibody levels across ten passages, as confirmed by Western blot and Protein A Octet analysis. Product-quality analysis by size-exclusion chromatography and reducing SDS-PAGE confirmed formation of properly assembled, mainly monomeric antibodies in both cases. Quantitative PCR indicated greater transgene copy numbers in MAR pools (+ 48% for the light chain and + 71% for the heavy chain), and RT-qPCR showed roughly fourfold higher transcript levels for both chains relative to controls. Bioinformatic analysis revealed several SATB1 binding motifs within the MAR sequence, and ChIP-qPCR demonstrated SATB1 association with the MAR-linked transgene locus. Overall, the data suggested that a CHO-native MAR could enhance transgene dosage and transcriptional activity, while preserving product integrity, possibly through SATB1-mediated chromatin organization. Ongoing work includes chromatin-mark profiling and process-level productivity measurements to better define the impact of MAR-based vector design on biomanufacturing performance.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3625-3640"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817745","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":"Cloning, Transformation, and Reporter Gene Analysis of the SalT Promoter in Barley (Hordeum vulgare).","authors":"Zahid Abbas Malik, Jochen Kumlehn, Sabir Hussain Shah, Zeshan Hassan, Götz Hensel, Nasir Ahmad Saeed","doi":"10.1007/s12033-026-01570-5","DOIUrl":"10.1007/s12033-026-01570-5","url":null,"abstract":"<p><p>Constitutive gene expression can lead to pleiotropic effects. Therefore, spatial or temporal restriction of expression via specific promoters provides a more targeted approach. This study aimed to clone the SalT promoter and analyze its activity in transgenic barley using GFP and GUS reporter genes. The T-DNA constructs carrying the SalT promoter were introduced into barley cv. Golden Promise, and transgenic plants were confirmed through PCR, hygromycin selection, and Southern hybridization. Both constructs, SalT-GFP and SalT-GUS, were transformed in barley cv. Golden Promise. Here, we characterized the expression pattern of the SalT promoter in barley and utilized it to drive the expression of reporter genes GFP and GUS. The SalT promoter was isolated from rice genomic DNA, cloned into the pNos-AB-M vector, and confirmed through PCR and restriction analysis. Subsequently, GFP and GUS genes were cloned under the SalT promoter in the same vector. The constructs were then subcloned into the p6U vector for plant expression. Agrobacterium-mediated genetic transformation of barley cultivar \"Golden Promise\" was conducted, resulting in successful integration of the transgenes. Callus induction, regeneration, and root formation efficiency were assessed, demonstrating the potential of the SalT promoter to drive gene expression during various stages of plant development. Molecular analyses, including PCR and Southern hybridization, confirmed the presence and integration of transgenes in the barley genome. Furthermore, GFP fluorescence and GUS staining analyses revealed strong expression of the respective genes under control of the SalT promoter in different plant tissues. This study provides insights into the application of the SalT promoter for genetic manipulation and functional characterization in barley, offering opportunities for crop improvement and biotechnological applications.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3607-3624"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817721","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 Cloning, Recombinant Expression, and In Silico Structural Analysis of Cu/Zn-Superoxide Dismutase from Trachyspermum ammi.","authors":"Lubna Siddiqui, Deepika Sharma, Shubhangi Pandey, Seneha Santoshi, Meenakshi Gupta, Maryam Sarwat, Alok K Sinha, Nidhee Chaudhary","doi":"10.1007/s12033-026-01584-z","DOIUrl":"10.1007/s12033-026-01584-z","url":null,"abstract":"<p><p>Superoxide dismutase (SOD) is an essential antioxidant metalloenzyme that is critical for the cellular defense against oxidative damage, as it scavenges superoxide radicals and maintains the redox status. Cytosolic Cu/Zn-SOD is particularly important in the regulation of oxidative stress among different isoforms in higher plants. While Cu/Zn-SODs from several plant species have been characterized, molecular information is limited for Trachyspermum ammi, a medicinally important member of a family Apiaceae with antioxidant potential.In the present study, an integrated molecular and in silico approach has been taken to clone and analyze a Cu/Zn type SOD gene from T. ammi to get insight into its structural and evolutionary characteristics. PCR amplification yielded an open reading frame of 456 bp encoding a protein of 152 amino acids. Sequence analysis showed that plant Cu/Zn-SODs, especially those from Daucus carota, were highly similar to one another (about 90-95%).Multiple sequence alignment confirmed the presence of conserved catalytic motifs and metal-binding histidine residues, both of which are crucial for enzymatic function. Physicochemical analysis predicted the protein to be stable, hydrophilic and compatible with cytosolic localization. The analysis of secondary structure indicated a predominance of β-strands, consistent with the conserved β-barrel architecture of plant Cu/Zn-SODs.The three-dimensional structure was built by homology modeling using a closely related plant Cu/Zn-SOD template with high sequence identity. Structural validation demonstrated an acceptable stereochemical quality with 86.3% residues in the favored region of Ramachandran plot, satisfactory ERRAT and Verify3D scores, and a low RMSD value of 0.104 Å on structural superimposition. Phylogenetic analysis placed the enzyme in the Apiaceae lineage, suggesting evolutionary conservation among related plant species. In conclusion, this study presents the first molecular and structural characterization of Cu/Zn-SOD from T. ammi and confirms the existence of a conserved structural framework typical of plant Cu/Zn-SODs. These results provide a basis for further studies concerning recombinant expression, enzymatic validation and potential relevance in antioxidant and plant stress biology.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3700-3714"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148138644","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":"Nanostructured Materials for Drug Delivery and Tissue Engineering Applications.","authors":"Antonela Matić, Emina Karahmet Sher, Esma Karahmet Farhat, Farooq Sher","doi":"10.1007/s12033-023-00784-1","DOIUrl":"10.1007/s12033-023-00784-1","url":null,"abstract":"<p><p>Nanotechnology and nanostructured materials for drug delivery and tissue engineering applications are relatively new field that is constantly advancing and expanding. The materials used are at the nanoscale level. Recently, great discoveries and applications have been made (Agents for use in chemotherapy, biological agents and immunotherapy agents) in the treatment of diseases in various areas. Tissue engineering is based on the regeneration and repair of damaged organs and tissues by developing biological substitutes that restore, maintain or improve the function of tissues and organs. Cells isolated from patients are used to seed 3D nanoparticles that can be synthetic or natural biomaterials. For the development of new tissue in tissue engineering, it is necessary to meet the conditions for connecting cells. This paper will present the ways of connecting cells and creating new tissues. Some recent discoveries and advances in the field of nanomedicine and the application of nanotechnology in drug delivery will be presented. Furthermore, the improvement of the effectiveness of new and old drugs based on the application of nanotechnology will be shown.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3473-3490"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13558372/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10029847","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":"Genome-Wide Identification and Characterization of Thaumatin-Like Proteins in Potato (Solanum tuberosum L.) and Their Role in Stress Tolerance.","authors":"Aiana, Garima Kakkar, Tanvi Mongia, Santosh Kumar Upadhyay, Kashmir Singh","doi":"10.1007/s12033-026-01569-y","DOIUrl":"10.1007/s12033-026-01569-y","url":null,"abstract":"<p><p>Thaumatin-like proteins (TLPs), part of the Pathogenesis-related protein 5 (PR5) family, play key roles in plant defense against biotic and abiotic stresses. In Solanum tuberosum, a crucial global food crop, the functional diversity of TLPs under stress conditions remains poorly understood, hindering efforts to improve stress tolerance. This study aimed to address this gap by performing a genome-wide identification and characterization of the TLP gene family in potato. We identified 34 TLPs (StTLP1 to StTLP34), distributed across 11 chromosomes. Detailed analyses were conducted on their physicochemical properties, gene structures, conserved motifs, and expression patterns. Promoter analysis revealed multiple stress-responsive cis-elements. Differential expression analysis showed that several StTLPs are significantly regulated in response to salinity, heat, and pathogen infection. Protein-protein interaction and miRNA targeting analyses further highlighted the regulatory networks involving StTLPs in stress adaptation. This study advances the theoretical understanding of the roles of StTLPs in stress response. It provides a valuable genetic resource for future efforts to enhance stress resilience in potato, with potential applications in crop improvement strategies. The 0 l expression of three TLP genes under salt and mannitol stress was verified through real-time PCR analysis after the interval of 15 days. Alterations in the expression patterns of StTLPs offered deeper insight into the involvement of this gene family in diverse abiotic stress responses. All three StTLPs were upregulated under both treatments at 400 mM, relative to their expression at 200 mM. The highest level of upregulation was observed in StTLP20, indicating its prominent role in both stress treatments.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3572-3590"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147468650","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":"Gene Cloning, Expression, and Purification of Kunitz Trypsin Inhibitor from Glycine max Using Halo Tag.","authors":"Jemy Jacob, Kannan Vadakkadath Meethal","doi":"10.1007/s12033-026-01583-0","DOIUrl":"10.1007/s12033-026-01583-0","url":null,"abstract":"<p><p>Soybean Kunitz Trypsin Inhibitor (SKTI) is one of the most extensively studied protease inhibitors, with applications in pest management, medicine, the food processing industry, and the leather industry. In this study, SKTI was cloned into the pFN29A Flexi vector containing a barnase gene. Genomic DNA was isolated from tender soybean leaves, and SKTI was amplified by PCR to obtain a 671 bp product. After cloning, an internal 380 bp sequence was amplified using specific primers to confirm that the cloned sequence was a functional SKTI, as non-functional SKTI genes also exist in Glycine max. The amplified PCR product, containing an AsiSI site at the 5' end and a PmeI site at the 3' end, was cloned into the pFN29A vector. The resulting colonies were screened by colony PCR, and the insert sequence was confirmed by Sanger sequencing. The recombinant protein, containing a His-tag, Halo-tag, and a TEV protease cleavage site, was expressed in Escherichia coli BL21 cells. Maximum expression was achieved 5 h after induction with 0.5 mM IPTG at 37 °C. The expressed SKTI was purified using affinity chromatography on HaloLink resin, and the bound SKTI was cleaved with HaloTEV protease to obtain pure SKTI. The purified inhibitor effectively inhibited bovine trypsin, with an IC₅₀ of 0.6 ± 0.003 µg/µl, yielding 1.6 mg per gram of bacterial pellet. The 24 kDa inhibitor remained stable up to a temperature of 50 °C. Kinetic analysis revealed that recombinant SKTI competitively inhibits trypsin, with a Kᵢ value of 14 µM.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3688-3699"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148043078","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":"Assessing Microalgal Protein's Impact on Environment and Energy Footprint via Life Cycle Analysis.","authors":"Hemamalini Rawindran, Jun Wei Lim, Man Kee Lam, Uganeeswary Supramaniam, Woei-Yenn Tong, Hui-Suan Ng, Muhammad Kashif Shahid, Chuxia Lin, Anwar Usman, Mohammad Shahzad Samdani","doi":"10.1007/s12033-023-00955-0","DOIUrl":"10.1007/s12033-023-00955-0","url":null,"abstract":"<p><p>Conventionally, increasing the yield of microalgal biomass has been the primary focus of research, while the significant protein reserve within this biomass has remained largely unexplored. This protein reserve possesses substantial value and versatility, offering a wide range of prospective applications and presenting an enticing chance for innovation and value enhancement for various sectors. Current study employed an innovative research approach that focused solely on the LCA of protein production potential from microalgal biomass, a lesser-explored aspects within this domain. Most environmental impact categories were shown to be significantly affected by cultivation phase because of the electrical obligation, followed by the harvesting and protein extraction phase. Still, the environmental aspect was seen to yield a minimal impact on global warming potential, i.e., 4 × 10^-3 kg CO₂, underscoring the ecologically favorable nature of the process. Conversely, the overall energy impact was seen to intensify with NEB of - 39.33 MJ and NER of 0.49, drawing attention to the importance of addressing the energy aspect to harness the full potential of microalgal protein production.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3765-3774"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"107591773","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 Study of Enzymatic Degradation of Bioplastic by Microalgae: An Outlook on Microplastic Environmental Impact Assessment, Challenges, and Opportunities.","authors":"Nikita Sharma, Suneetha Vuppu","doi":"10.1007/s12033-023-00886-w","DOIUrl":"10.1007/s12033-023-00886-w","url":null,"abstract":"<p><p>Microplastics are tiny pieces of non-biodegradable plastic that can take thousands of years to break down. As microplastics degrade, they release harmful compounds into the environment, which can be found in the surroundings. The microplastics found in the environment are hard to detect and remove because of their small particle sizes. Microplastics cannot decompose naturally, so they accumulate in the environment and cause pollution. As a result, bioplastics can be produced from a vast array of substrates, including biopolymers, citrus peels, leather, and feather wastes. Blue-green microalgae namely Arthrospira platensis (spirulina) contains enzymes such as laccase and catalase which can be responsible for the degradation of bioplastics. In our study, we performed molecular docking to identify the binding affinities of different enzymes such as laccase and catalase with different substrates, focusing on determining the most suitable substrate for enhancing enzyme activity for degradation of bioplastics. The analysis revealed that veratryl alcohol is the most suitable substrate for laccase, whereas lignin is the more preferred substrate for catalase with the highest binding affinity score of - 5.9 and - 8.1 kcal/mol. Moreover, degradation, challenges, opportunities, and applications of bioplastics in numerous domains such as cosmetics, electronics, agriculture, medical, textiles, and food industries have also been highlighted.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3523-3553"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41135540","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":"Organic Molecules are Involved in Ni-Struvite Biomineralization by Streptomyces mirabilis.","authors":"Flávio Silva Costa, Thomas Krüger, Nico Ueberschaar, Olaf Kniemeyer, Axel Brakhage, Erika Kothe","doi":"10.1007/s12033-026-01580-3","DOIUrl":"10.1007/s12033-026-01580-3","url":null,"abstract":"<p><p>The Gram-positive, filamentous soil bacterium Streptomyces mirabilis P16B-1 has been shown to tolerate high environmental concentrations of Ni. The extremely high metal tolerance is partly provided through biomineralization, observed for both Mg bearing struvite and Ni-struvite where Ni ions replace Mg in the mineral lattice. The minerals showed different morphologies. This led us to assume that excreted substances may have the capacity to influence the mineral formation process that occurs at a distance to the cells. Here, we could show with metabolomics and proteomics studies that 23 metabolites as well as 18 proteins potentially co-precipitate with the minerals. Secreted secondary metabolites reduced in the supernatant after mineralization were identified as ergothioneine, diacetyllegionaminic acid, nevaltophin F, and diaminonaphthalene. The compounds phenylacetaldehyde and margaric acid also reduced after mineral precipitation did not alter mineral macromorphology. In addition, ten proteins that co-precipitated with statistical significance were predicted to be secreted. Among those, a specific nickel binding protein, NikA, was reduced in high amounts from the supernatant upon crystal formation. The potential of secreted organic molecules and proteins to bind specific surfaces of the mineral thereby redirecting crystal growth in a microbially influenced biomineralization process. Implications for biotechnological metal-struvite use are discussed.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"3659-3673"},"PeriodicalIF":3.2,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13558439/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147948923","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}