Journal of biotechnology最新文献_第8页

Corrigendum to “Substitution of reactive centre loop residues from C1 esterase inhibitor increases the inhibitory specificity of alpha-1 antitrypsin for plasma kallikrein” [J. Biotechnol. 405 (2025) 205–214] 从C1酯酶抑制剂中取代反应性中心环残基增加α -1抗胰蛋白酶对血浆钾激肽的抑制特异性[J]。生物工程学报，2005(5):389 - 389。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2026-01-01 Epub Date: 2025-11-08 DOI: 10.1016/j.jbiotec.2025.11.003

Sangavi Sivananthan , S. Ameer Ahmed , Ammaar M. Baig , Varsha Bhakta , William P. Sheffield

引用次数: 0

Beyond tyrosine feeding: A novel fed-batch cultivation strategy based on tyrosine metabolic engineering in recombinant CHO cells 超越酪氨酸饲养：一种基于酪氨酸代谢工程的重组CHO细胞补料分批培养新策略。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2026-01-01 Epub Date: 2025-10-10 DOI: 10.1016/j.jbiotec.2025.10.003

Lei Cao , Liang Zhao , Qian Ye , Wen-Song Tan

{"title":"Beyond tyrosine feeding: A novel fed-batch cultivation strategy based on tyrosine metabolic engineering in recombinant CHO cells","authors":"Lei Cao , Liang Zhao , Qian Ye , Wen-Song Tan","doi":"10.1016/j.jbiotec.2025.10.003","DOIUrl":"10.1016/j.jbiotec.2025.10.003","url":null,"abstract":"<div><div>CHO cells dominate monoclonal antibody (mAb) production in fed-batch biomanufacturing, where tyrosine supply is limited by low solubility in neutral media and the complexities of alkaline tyrosine feeds. Existing studies confirm overexpressing pterin-4α-carbinolamine dehydratase 1 (PCBD1) and phenylalanine hydroxylase (PAH) restores tyrosine prototrophy and matches non-engineered cells’ production in tyrosine-supplemented cultures. However, these studies focus on enzyme regulation without resolving feeding-phase supply challenges in high-density scenarios. To address this, this study verified that recombinant CHO (rCHO) cells rely strongly on exogenous tyrosine for growth and production. Multi-level expression analysis further confirmed low PCBD1/PAH levels restrict endogenous tyrosine synthesis, and identified quinoid dihydropteridine reductase (QDPR)—a key tetrahydrobiopterin (BH4) regeneration enzyme—as a previously unrecognized bottleneck, particularly in tyrosine-limited conditions. By co-overexpressing QDPR in high PCBD1/PAHexpressing cells to remodel the tyrosine biosynthesis pathway, a novel fed-batch strategy was established: basal medium with 3.0 mM tyrosine and tyrosine-free singlefeeding medium. Results showed this strategy, effective in high-density fed-batch settings, enabled rCHO cells to reach a final mAb titer of 4.24 g/L, representing a 32.50% increase compared to cells overexpressing only PCBD1 and PAH, and a 10.70 % increase compared to the conventional strategy. In summary, the strategy offers a simplified nutrient alternative by eliminating alkaline tyrosine feeds, highlighting holistic metabolic pathway optimization’s importance in biomanufacturing and targeted value for high-density, tyrosine-limited CHO cell-based mAb production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 22-32"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145279962","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-concentration Mg2+ stress improves L-lactic acid biosynthesis of Bacillus coagulans revealed by combined analysis of transcriptome and metabolome 转录组和代谢组联合分析表明高浓度Mg2+胁迫促进了凝结芽孢杆菌l -乳酸的生物合成。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2026-01-01 Epub Date: 2025-09-30 DOI: 10.1016/j.jbiotec.2025.09.016

Zhitong Sun , Jiaxin Liang , Gang Wang , Yumeng Zhen , Jinlong Liu , Di Cai , Bin Wang , Yong Wang

{"title":"High-concentration Mg2+ stress improves L-lactic acid biosynthesis of Bacillus coagulans revealed by combined analysis of transcriptome and metabolome","authors":"Zhitong Sun , Jiaxin Liang , Gang Wang , Yumeng Zhen , Jinlong Liu , Di Cai , Bin Wang , Yong Wang","doi":"10.1016/j.jbiotec.2025.09.016","DOIUrl":"10.1016/j.jbiotec.2025.09.016","url":null,"abstract":"<div><div>The application of magnesium oxide as a neutralizing agent significantly enhanced L-lactic acid (L-LA) production in <em>Bacillus coagulans</em> (<em>B. coagulans</em>), increasing titer, yield, productivity, and cell viability by 21.81 %, 7.61 %, 22.22 %, and 18.50 times, respectively. To elucidate the metabolic response to high-concentration Mg<sup>2+</sup> stress, transcriptomic analysis identified 1021 differentially expressed genes (DEGs), with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealing predominant biological processes and pathway dysregulation. Metabolomic profiling detected 1196 differential metabolites, of which 216 were annotated to 129 metabolic pathways. Integrated multi-omics analyses delineated 10 core pathways involving 139 DEGs and 29 metabolites, demonstrating that Mg<sup>2+</sup> stress enhances lactic acid fermentation efficiency through reprogramming of carbon flux, nucleic acid biosynthesis, amino acid metabolism, cofactor dynamics, membrane transport, and transcriptional regulation. This study provides mechanistic insights into <em>B. coagulans</em> adaptation to Mg<sup>2+</sup> stress and proposes a viable strategy to optimize industrial L-LA bioproduction.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 1-13"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145212742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Morphology and bioproduction potential of DCB-induced cell wall-deficient tobacco BY-2 cells and transcriptomic analysis dcb诱导的烟草BY-2细胞壁缺陷细胞形态、生物生产潜力及转录组学分析。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2026-01-01 Epub Date: 2025-11-05 DOI: 10.1016/j.jbiotec.2025.11.007

Uddhab Karki , Shekoofeh Sadravi , Katelin Kellar , Marin Hirata , Fnu Mamta , Jianfeng Xu

{"title":"Morphology and bioproduction potential of DCB-induced cell wall-deficient tobacco BY-2 cells and transcriptomic analysis","authors":"Uddhab Karki , Shekoofeh Sadravi , Katelin Kellar , Marin Hirata , Fnu Mamta , Jianfeng Xu","doi":"10.1016/j.jbiotec.2025.11.007","DOIUrl":"10.1016/j.jbiotec.2025.11.007","url":null,"abstract":"<div><div>Plant cell cultures, especially tobacco BY-2 cells, represent a promising platform for recombinant protein production. However, their full potential is hindered by several challenges. Some of these challenges, such as inefficient protein secretion, the presence of large vacuoles, and difficulties in cryopreservation can be attributed to the rigidity of the plant cell wall. We developed and characterized cell wall-deficient BY-2 cells by gradually adapting them to the cellulose biosynthesis inhibitor 2,6-dichlorobenzonitrile (DCB). The DCB-adapted cells exhibited marked morphological changes, including cell clumping due to pectin overaccumulation and an ∼80 % reduction in cellulose content. Despite these alterations, the DCB-adapted BY-2 cells maintained robust growth and demonstrated efficient <em>Agrobacterium</em>-mediated transformation. The production and secretion levels of the model protein EGFP and the therapeutic protein human EPO were 2.1- to 2.8-fold higher than those observed in native BY-2 cells. However, secretion of EGFP fused with a hydroxyproline-<em>O</em>-glycosylated module (SP)₃₂ was dramatically enhanced by up to 35-fold. Transcriptomic analysis revealed broad gene expression changes, including down-regulation of genes involved in cellulose biosynthesis and upregulation of genes associated with branched pectic polysaccharide synthesis. These results provide a proof of concept that cell wall-deficient plant cell lines can serve as effective platforms for recombinant protein production, laying foundation for further engineering of plant cells as next-generation biofactories.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 170-181"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145471132","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Semi-rational design of the substrate-binding pocket of acyl-CoA dehydrogenase Tfu_1647 to improve catalytic activity for 6-carbon substrates 半合理设计酰基辅酶a脱氢酶Tfu_1647底物结合袋，提高对6碳底物的催化活性。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2026-01-01 Epub Date: 2025-10-27 DOI: 10.1016/j.jbiotec.2025.10.007

Lixia Liu , Shenghu Zhou , Yu Deng

{"title":"Semi-rational design of the substrate-binding pocket of acyl-CoA dehydrogenase Tfu_1647 to improve catalytic activity for 6-carbon substrates","authors":"Lixia Liu , Shenghu Zhou , Yu Deng","doi":"10.1016/j.jbiotec.2025.10.007","DOIUrl":"10.1016/j.jbiotec.2025.10.007","url":null,"abstract":"<div><div><em>Tfu</em>_1647, an acyl-CoA dehydrogenase, reversible catalyses the conversion of enoyl-CoA to acyl-CoA and is often utilized in the biosynthesis of adipic acid. However, its applicability constrained by a lack of three-dimensional structural information and its broad substrate acceptance. In this study, we demonstrate structural and biochemical investigations of 6-carbon substrate (6C-CoA) dehydrogenase <em>Tfu</em>_1647 with high-efficiency. Notably, <em>Tfu</em>_1647 displayed a high substrate affinity for the intermediate-length acyl-CoA, which is an essential precursor for the synthesis of fatty acids. A semi-rational design approach, informed by the crystal structure, was employed to enhance the specific activity of 6C-CoA by identifying effective variants. Within this cohort of variants, <em>Tfu</em>_1647<sup>Thr370Gln(T370Q)</sup> demonstrated the highest affinity for 6C-CoA (<em>K</em><sub>d</sub> = 1.48 × 10⁻⁶ M<sup>−1</sup>) and the greatest enzyme activity (<em>k</em><sub>cat</sub> = 0.98 min⁻¹), representing a 25.7 % improvement in binding affinity and a 2.3-fold increase in catalytic rate compared to wild-type <em>Tfu</em>_1647. A comprehensive comparison revealed that the <em>Tfu</em>_1647<sup>T370Q</sup> enlarges the substrate pocket and subtly alters local electrostatics, thereby preserving FAD binding while improving substrate affinity (25.7 % lower <em>K</em><sub>d</sub>) and catalytic efficiency (3.7-fold higher <em>k</em><sub>cat</sub>/<em>K</em><sub>M</sub>) toward 6C-CoA. Molecular dynamics and free energy analyses further showed that this substitution refines substrate specificity, conferring a distinct preference for medium-chain (C6) substrates over shorter or longer acyl-CoAs. This finding paves the way for the rational design of <em>Tfu</em>_1647 for its application in adipic acid biosynthesis.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"409 ","pages":"Pages 96-106"},"PeriodicalIF":3.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145400788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhancement and mechanism of in-cluster two-component regulatory factors on granaticin production in Streptomyces vilmorinianum YP1 簇内双组分调控因子对vilmorinianum Streptomyces YP1 Granaticin生产的增强作用及其机制

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-12-01 Epub Date: 2025-08-30 DOI: 10.1016/j.jbiotec.2025.08.015

Bingyu Zhao , Qiaolei Zhu , Xin Wang , Xiaoya Shang , Yunping Zhu

{"title":"Enhancement and mechanism of in-cluster two-component regulatory factors on granaticin production in Streptomyces vilmorinianum YP1","authors":"Bingyu Zhao , Qiaolei Zhu , Xin Wang , Xiaoya Shang , Yunping Zhu","doi":"10.1016/j.jbiotec.2025.08.015","DOIUrl":"10.1016/j.jbiotec.2025.08.015","url":null,"abstract":"<div><div>Granaticins, a class of bioactive benzoisochromanequinones (BIQs), are natural blue pigments with biological activities, including antibacterial properties and promising clinical anticancer applications. However, their clinical and industrial applications are limited by low production yields and unclear biosynthetic regulation. In this study, we identified a rare in-cluster two-component system (TCS; <em>orf10</em>/<em>orf11</em>) within the granaticin biosynthesis-related gene cluster (BGC) of <em>Streptomyces vilmorinianum</em> YP1, a novel high-yielding producer. The <em>orf10</em> encodes a MerR family transcription regulator, while the <em>orf11</em> encodes a sensor histidine kinase. The roles of <em>orf10</em> and <em>orf11</em> were determined using single and double knockouts and complementation experiments. The yield of granaticins increased with the overexpression of the two genes in <em>S. vilmorinianum</em> YP1. Notably, <em>orf10</em> overexpression resulted in a record high granaticin B titer of 716.27 mg/L, representing a substantial improvement over native production level. The metabolic map obtained using Ultra Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS) analysis revealed that the metabolic processes of the combined overexpression mutant were significantly different from those of the single overexpression mutants. These findings not only establish the functional link between the in-cluster TCSs, and granaticin biosynthesis but also provide an effective engineering strategy to overcome production bottlenecks. Our work advances the mechanistic understanding of BIQ regulation and highlights <em>S. vilmorinianum</em> YP1 as a promising platform for industrial-scale granaticin production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 39-49"},"PeriodicalIF":3.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144955211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Improving thermostability of α-L-fucosidase from Pedobacter sp. via consensus-guided engineering and directed evolution 通过共识导向工程和定向进化提高Pedobacter sp. α-L- focusidase的热稳定性。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-12-01 Epub Date: 2025-09-22 DOI: 10.1016/j.jbiotec.2025.09.011

Tong-Tong Zhao , Yu-Bo Wang , Wen-Qian Qiu , Ying-Ying Wang , Yang-Yong Lv , Huan-Chen Zhai , Yuan-Sen Hu , Zheng-Qiang Jiang , Shuai-Bing Zhang

{"title":"Improving thermostability of α-L-fucosidase from Pedobacter sp. via consensus-guided engineering and directed evolution","authors":"Tong-Tong Zhao , Yu-Bo Wang , Wen-Qian Qiu , Ying-Ying Wang , Yang-Yong Lv , Huan-Chen Zhai , Yuan-Sen Hu , Zheng-Qiang Jiang , Shuai-Bing Zhang","doi":"10.1016/j.jbiotec.2025.09.011","DOIUrl":"10.1016/j.jbiotec.2025.09.011","url":null,"abstract":"<div><div>The α-L-fucosidase from <em>Pedobacter</em> sp. (<em>Pb</em>Fuc), a glycoside hydrolase capable of catalyzing the synthesis of 2’-fucosyllactose (2’-FL) from 4-nitrophenyl α-L-fucopyranoside (<em>p</em>NP-Fuc) and D-lactose, exhibits limited industrial applicability due to inherent thermostability constraints. This study implemented a combinatorial strategy integrating consensus-guided engineering and directed evolution to engineer the thermal stability of <em>Pb</em>Fuc, resulting in the identification of six critical mutants (G53C, M54I, N59S, T71H, R125C, S199P) and the subsequent construction of the combinatorial mutant M6. Thermostability assays revealed complete inactivation of the wild-type enzyme after 30-min incubation at 45 °C, whereas M6 retained approximately 40 % residual activity under equivalent conditions at 60 °C, accompanied by an increase in the optimal reaction temperature from 35 °C to 40 °C. Structural mechanism analysis demonstrated that the enhanced thermostability of M6 originated from synergistic multilevel structural optimization and reorganization of molecular interaction networks: Conformational stabilization manifested through prolonged maintenance of stable secondary structural conformations during thermal stress and reduced amplitude of tertiary structural fluctuations; Global structural compaction decreased solvent-accessible surface area, thereby minimizing thermal energy transfer; Local structural reinforcement occurred via the formation of novel hydrogen bonds, enhanced rigidity through π-π stacking, and neutralization of electrostatic repulsion via charge compensation.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 168-180"},"PeriodicalIF":3.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145137638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Komagataella phaffii as a microbial cell factory for antimicrobial peptide production Komagataella phaffii作为抗菌肽生产的微生物细胞工厂。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-12-01 Epub Date: 2025-09-30 DOI: 10.1016/j.jbiotec.2025.09.015

Michel Lopes Leite , Kamila Botelho Sampaio de Oliveira , Letícia Ferreira Lima , Nadielle Tamires Moreira Melo , José Brango-Vanegas , Hugo Costa Paes , Octávio Luiz Franco

{"title":"Komagataella phaffii as a microbial cell factory for antimicrobial peptide production","authors":"Michel Lopes Leite , Kamila Botelho Sampaio de Oliveira , Letícia Ferreira Lima , Nadielle Tamires Moreira Melo , José Brango-Vanegas , Hugo Costa Paes , Octávio Luiz Franco","doi":"10.1016/j.jbiotec.2025.09.015","DOIUrl":"10.1016/j.jbiotec.2025.09.015","url":null,"abstract":"<div><div>In recent decades, the methylotrophic yeast <em>Komagataella phaffii</em> has emerged as a powerful host for the heterologous production of antimicrobial peptides (AMPs) as an economical and scalable platform. <em>K. phaffii</em> combines several important advantages for a recombinant expression system, such as rapid growth, high-density cell culture, efficient protein secretion, and the ability to perform essential post-translational modifications. The methanol-inducible AOX1 promoter (P<sub><em>AOX1</em></sub>), generally employed through the pPICZalpha vector, allows strong and tightly regulated heterologous expression and is a central factor in the expression system of this yeast. However, several strategies have been employed to produce recombinant AMPs in <em>K. phaffii</em>, among them codon optimization, engineered methanol-resistant strains, alternative promoters, and the use of different secretory peptides. This review highlights the latest advances and practical considerations in the use of <em>K. phaffii</em> for AMP production, discussing challenges such as peptide stability, proteolytic degradation, and yield optimization. The insights provided contribute to the expansion of biotechnological applications of <em>K. phaffii</em>, reinforcing its potential as an efficient and safe platform for large-scale production of therapeutic AMPs.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 275-287"},"PeriodicalIF":3.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145212745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Chitosan application in cosmetics and dermatology – Antimicrobial and prebiotic potential to control human microbiome 壳聚糖在化妆品和皮肤病学中的应用-抗菌和益生元控制人体微生物群的潜力。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-12-01 Epub Date: 2025-09-27 DOI: 10.1016/j.jbiotec.2025.09.014

Masha Delic, Irina Butorova, Andrey Kuskov

{"title":"Chitosan application in cosmetics and dermatology – Antimicrobial and prebiotic potential to control human microbiome","authors":"Masha Delic, Irina Butorova, Andrey Kuskov","doi":"10.1016/j.jbiotec.2025.09.014","DOIUrl":"10.1016/j.jbiotec.2025.09.014","url":null,"abstract":"<div><div>The human microbiome has become a critically important field of study, with recent advances continuously revealing new aspects of its functional roles. A deeper understanding of the complex metabolic interactions between the microbiome, therapeutic agents, and the human host is now developing. A key challenge is the development of effective novel antimicrobial compounds, particularly those targeting biofilm-related infections implicated in a wide range of chronic diseases, to better control the human microbiota and treat typical skin diseases. Chitosan, a polysaccharide derived from various natural sources, possesses a unique set of biological properties, making it a promising base for numerous cosmetic and medicinal products aimed at managing dermatological diseases and modulating the microbiome. By altering its characteristics, chitosan's properties can be tuned to produce biomacromolecules with optimized activity beneficial to human microbiota. This review summarizes the sources and mechanisms underlying chitosan's antimicrobial activity, its immunomodulatory and anti-pathogenic effects, and critically evaluates its dual potential as both an antimicrobial and a prebiotic agent. Special emphasis is placed on chitosan-based formulations for treating skin diseases and promoting microbiome health – an emerging and promising area of research.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 217-231"},"PeriodicalIF":3.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145191686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A next-generation platform for highly optimized CRISPR-mediated transcriptional repression 高度优化crispr介导的转录抑制的下一代平台。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-12-01 Epub Date: 2025-09-24 DOI: 10.1016/j.jbiotec.2025.09.012

Andrew Kristof , Krithika Karunakaran , Yann Ferry , Paula Mizote , Christopher Allen , Sophie Briggs , John Blazeck

{"title":"A next-generation platform for highly optimized CRISPR-mediated transcriptional repression","authors":"Andrew Kristof , Krithika Karunakaran , Yann Ferry , Paula Mizote , Christopher Allen , Sophie Briggs , John Blazeck","doi":"10.1016/j.jbiotec.2025.09.012","DOIUrl":"10.1016/j.jbiotec.2025.09.012","url":null,"abstract":"<div><div>CRISPR interference (CRISPRi), the fusion of transcriptional repressor domains to nuclease-inactive Cas9, is a powerful genetic tool enabling site-specific suppression of gene expression. However, its performance remains inconsistent across cell lines, gene targets, or single guide RNAs (sgRNAs) employed. This study described the development process of a novel, highly optimized CRISPRi repressor for mammalian gene regulation through a multi-pronged protein engineering approach: (1) truncating established domains, (2) characterizing candidate domains, (3) creating combinatorial domain fusion libraries, and (4) optimizing NLS configuration. First, by evaluating several truncations of MeCP2, a well-established repressor, we see that the ultra-compact NCoR/SMRT interaction domain (NID) significantly enhances CRISPRi gene knockdown performance, exceeding levels observed with canonical MeCP2 subdomains by an average of ∼40 %. Incorporating this optimized MeCP2 NID truncation with a diverse panel of authenticated repressor domains, we next assemble and screen combinatorial multi-domain libraries, discovering four new repressor fusions. Upon follow-up nuclear localization signal (NLS) configuration analysis, we see that affixing one carboxy-terminal NLS enhances gene knockdown efficiency of the repressors by an average of ∼50 %. Through rigorous validation of NLS-tagged repressor fusions across several cell lines, multiple sgRNA targets, and genome-wide dropout screens, we establish that our strongest system, dCas9-ZIM3-NID-MXD1-NLS, achieves superior gene silencing capabilities over alternative CRISPRi platforms. In addition to developing dCas9-ZIM3-NID-MXD1-NLS, a uniquely potent transcriptional repressor, we envision that the multi-domain engineering approach utilized in this study will be valuable framework enabling future strides in CRISPR platform development.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 192-200"},"PeriodicalIF":3.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145175621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0