New biotechnology最新文献_第3页

More than meets the eye: microalgal-bacterial association in polyhydroxybutyrate (PHB) accumulating cultures of Desmodesmus communis 超过满足眼睛：微藻-细菌联合在聚羟基丁酸（PHB）积累培养的桥丝霉。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2026-01-08 DOI: 10.1016/j.nbt.2026.01.005

Martina Franchini , Mara Simonazzi , Federica Costantini , Valentina Papa , Rossella Pistocchi , Stefano Ratti , Chiara Samorì , Laura Pezzolesi

{"title":"More than meets the eye: microalgal-bacterial association in polyhydroxybutyrate (PHB) accumulating cultures of Desmodesmus communis","authors":"Martina Franchini , Mara Simonazzi , Federica Costantini , Valentina Papa , Rossella Pistocchi , Stefano Ratti , Chiara Samorì , Laura Pezzolesi","doi":"10.1016/j.nbt.2026.01.005","DOIUrl":"10.1016/j.nbt.2026.01.005","url":null,"abstract":"<div><div>The focus on microalgae within a circular economy framework has been growing due to their ability to utilize inexpensive substrates and produce high-value biomolecules. Among these, the biopolymer polyhydroxybutyrate (PHB) can be accumulated as a storage compound in several microalgal biomasses under nutrient- and light-limiting conditions. Herein, the Chlorophyta <em>Desmodesmus communis</em> was grown under mixotrophic conditions (low light, phosphorus-free medium, 1 g L⁻¹ sodium acetate) in a 1 L semicontinuous system over 25 days. PHB accumulation progressively increased, reaching 57 % w/w on day 25. Since mixotrophic conditions also favor bacterial growth, the associated bacterial community was analyzed through 16S rRNA gene metabarcoding at different time points (day 0, 12, 21, 25) to investigate its potential contribution to PHB accumulation. A selection in bacterial genera was observed after prolonged cultivation. Moreover, antibiotic treatments markedly reduced bacterial diversity and PHB content, indicating a central bacterial role in PHB production and highlighting key genera likely involved. Environmental Scanning Electron Microscopy (ESEM) and Transmission Electron Microscopy (TEM) revealed close interactions between algal and bacterial cells, suggesting a potential endophytic presence of PHB-accumulating bacteria within <em>D. communis</em> cells. These findings propose a new perspective on microalgae-associated bacterial communities, viewing them not as contaminants but as essential contributors to microalgal growth and valuable metabolites production. The results emphasize the importance of considering the bacteria-microalgae consortium in PHB production research, rather than focusing solely on photoautotrophic organisms.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 192-200"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145949060","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

Automated optimization of resin selection, wash parameters, and elution parameters for protein A-based purifications 自动优化树脂选择，洗涤参数和洗脱参数的蛋白质为基础的纯化。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-11-11 DOI: 10.1016/j.nbt.2025.11.004

Gracie K. Anderson , Mark Hicks , Penny Hamlyn , L. Andrew Lee , Patrick A. Kates

{"title":"Automated optimization of resin selection, wash parameters, and elution parameters for protein A-based purifications","authors":"Gracie K. Anderson , Mark Hicks , Penny Hamlyn , L. Andrew Lee , Patrick A. Kates","doi":"10.1016/j.nbt.2025.11.004","DOIUrl":"10.1016/j.nbt.2025.11.004","url":null,"abstract":"<div><div>Automated protein purification is essential for meeting the growing demands of high-throughput sample processing capabilities, especially when screening multiple parameters. For instance, protein A-based affinity chromatography is an important downstream antibody purification process that can be automated to maintain high reproducibility with minimized human error. This purification step can clear undesirable host cell proteins (HCP) from Chinese hamster ovary (CHO) cells and capture target monoclonal antibodies, developed for a broad assortment of therapeutic applications, from antibody-expressing cell culture fluid. Selecting optimized parameters for protein A purification is crucial to ensuring productive performance in terms of optimal binding, prevention of undesirable effects from host cell protein, and product aggregate reduction. Herein, we demonstrate a faster and lower cost automated micro-purification workflow to accelerate the screening and selection of resin, wash, and elution parameters for protein A chromatography. In testing three monoclonal antibodies (mAb) and one bispecific monoclonal antibody (bsAb), we observed marked variability in purity and HCP clearance based on the wash buffer and elution pH. We also report that modified intermediate wash buffers containing sodium chloride minimized HCP contamination of eluted samples while preserving high monomer yields. Additionally, we identify differences in performance between resins, including the ability of Praesto Jetted A50 HipH to maintain higher yields at elution pH at which the yields of other resins declined (>pH 4.2). These results highlight the utility of automated microscale purification in reducing time and resource consumption involved in optimizing and executing downstream antibody bioprocessing.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 17-26"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145513561","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

Splint-free circular RNA synthesis via RNA secondary structure–guided ligation 通过RNA二级结构引导连接的无夹板环状RNA合成。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-12-11 DOI: 10.1016/j.nbt.2025.12.004

Yeong-Chan Kim , Dong Hyun Kang , Kanghyun Choi , Yeon-Jin Yoo , Hyun Jin Kim , Si-Sun Choi , Seung-Hoon Kang , Eung-Soo Kim

{"title":"Splint-free circular RNA synthesis via RNA secondary structure–guided ligation","authors":"Yeong-Chan Kim , Dong Hyun Kang , Kanghyun Choi , Yeon-Jin Yoo , Hyun Jin Kim , Si-Sun Choi , Seung-Hoon Kang , Eung-Soo Kim","doi":"10.1016/j.nbt.2025.12.004","DOIUrl":"10.1016/j.nbt.2025.12.004","url":null,"abstract":"<div><div>Circular RNA (circRNA) is a covalently closed RNA molecule in which the 5′ and 3′ ends are joined. CircRNAs can be generated via RNA circularization, a process that links the termini of linear RNA. The most common <em>in vitro</em> method for RNA circularization is the ribozyme-based permuted introns and exons (PIE) system. However, circRNAs produced by the PIE method retain partial exogenous exon sequences, potentially leading to immunogenicity issues. In this study, we developed an alternative approach that exploits the secondary structure of linear RNA and enzymatic ligation to synthesize circRNA <em>in vitro</em> for targeted gene expression. We first predicted RNA secondary structures and designed linear RNA molecules to form a terminal nick structure. Using T4 RNA ligase 2 (T4 Rnl2), we sealed the nick to connect the RNA ends. This ligase-mediated <em>in vitro</em> circularization achieved high efficiency and enabled functional expression of the encoded target gene. While ligase-based splint-free circularization has been described for small RNA circles, reports demonstrating functional protein expression from such constructs remain limited. This ligase-mediated RNA circularization approach should be an efficient alternative for production of circular RNA. This ligase-mediated, secondary-structure–guided, splint-free strategy using T4 RNA ligase 2 represents an efficient alternative for circRNA production, enabling both high-yield synthesis and validated gene expression in mammalian cells.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 120-126"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145752108","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

Transcriptional profiling of probiotic Escherichia coli Nissle 1917 under multiple environmental stresses: Simulated microgravity, microaerobic, and co-culture conditions 益生菌大肠杆菌Nissle 1917在多种环境胁迫下的转录谱分析：模拟微重力、微氧和共培养条件。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-11-22 DOI: 10.1016/j.nbt.2025.11.007

Jaewoo Yim , Sung Won Cho , Seongjun Park , Sukjae Han , Tae Hyun Kim , Sang Woo Seo

{"title":"Transcriptional profiling of probiotic Escherichia coli Nissle 1917 under multiple environmental stresses: Simulated microgravity, microaerobic, and co-culture conditions","authors":"Jaewoo Yim , Sung Won Cho , Seongjun Park , Sukjae Han , Tae Hyun Kim , Sang Woo Seo","doi":"10.1016/j.nbt.2025.11.007","DOIUrl":"10.1016/j.nbt.2025.11.007","url":null,"abstract":"<div><div>Environmental factors can substantially alter the physiology of probiotics, yet their transcriptomic responses under multiple simultaneous stresses remain poorly understood. Among these factors, a previous study has shown that the low-shear environment induced by simulated microgravity elicits mild stress in the well-known probiotic <em>Escherichia coli</em> Nissle 1917 (EcN). In this study, we investigated the impact of additional environmental stressors on the transcriptomic profile of EcN by introducing microaerobic conditions and co-culture with a pathogenic <em>Pseudomonas aeruginosa</em> PAO1 (PAO1), in combination with simulated microgravity.</div><div>Under the combined condition of simulated microgravity and microaerobic stress, EcN maintained stable growth while upregulating genes involved in anaerobic respiration and biofilm formation, indicating effective adaptation to oxygen-limited environments. However, when co-cultured with PAO1 under the same conditions, EcN exhibited pronounced growth inhibition. Transcriptomic analysis revealed downregulation of genes involved in energy metabolism and biofilm formation, while genes related to metal ion uptake remained active. Further comparison between simulated microgravity and normal gravity under these compound stress conditions revealed that microgravity further exacerbated growth suppression in the context of overlapping environmental stresses, particularly via altered amino acid metabolism.</div><div>These findings suggest that EcN’s stress responses and viability are impaired in complex environmental conditions, emphasizing the importance of developing more resilient probiotic strains for therapeutic applications in extreme environments such as spaceflight.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 62-73"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145596971","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

Investigating the dynamics of precursor availability for sesquiterpenoid production in the cytoplasm and plastid of Chlamydomonas reinhardtii 莱茵衣藻细胞质和质体中倍半萜类物质生产前体有效性的动态研究。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-11-17 DOI: 10.1016/j.nbt.2025.11.006

Malak N. Abdallah, Gordon B. Wellman, Sebastian Overmans, Kyle J. Lauersen

{"title":"Investigating the dynamics of precursor availability for sesquiterpenoid production in the cytoplasm and plastid of Chlamydomonas reinhardtii","authors":"Malak N. Abdallah, Gordon B. Wellman, Sebastian Overmans, Kyle J. Lauersen","doi":"10.1016/j.nbt.2025.11.006","DOIUrl":"10.1016/j.nbt.2025.11.006","url":null,"abstract":"<div><div>The green alga <em>Chlamydomonas reinhardtii</em> is an interesting organism for production of heterologous isoprenoid natural products through metabolic engineering as it can grow photosynthetically and represents a plant-like cell environment for synthase expression. Here, we investigated expression of the patchoulol synthase from <em>Pogostemon cablin</em> (Benth) (<em>Pc</em>PS) to probe the production of sesquiterpenoids from different sub-cellular compartments of the algal cell. We observed that knock-down of squalene synthase (SQS) in the cytoplasm did not affect sesquiterpene production titers in the plastid, suggesting no backward movement of FPP from cytoplasmic pools. Fusion of <em>Pc</em>PS with a farnesyl pyrophosphate synthase (FPPS) did not enhance patchoulol production in the cytoplasm but did in the plastid, as also reported elsewhere. Secondary transformations with cytoplasmic and chloroplast-localized <em>Pc</em>PS constructs increased synthase titers and improved patchoulol yields from both compartments. In multi-parallel photobioreactor experiments with patchoulol extraction through two-phase solvent contact, different daily patchoulol production behaviors were observed based on subcellular localization of the <em>Pc</em>PS and carbon source. Those with plastid localization of <em>Pc</em>PS-FPPS exhibited continued production into later stages of cultivation, while cytoplasm-localized <em>Pc</em>PS with SQS k.d. had the highest production rates in mid-logarithmic phase with low productivity in later growth stages. Cultivations in high-density photobioreactors with nutrient-enriched medium generated 15 mg patchoulol/L culture from CO<sub>2</sub>, the highest yield reported from the alga to date. The results indicate that the algal cell has flexible pools of isoprenoid precursors in different subcellular compartments from which the biotechnologist can design an expression strategy tailored to carbon-feeding regimes.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 52-61"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145557664","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

Integration of strain and process optimization to increase autotrophic growth of engineered Komagataella phaffii 菌株与工艺优化相结合促进工程法菲小松藻自养生长。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-11-27 DOI: 10.1016/j.nbt.2025.11.010

Michael Baumschabl , Lisa Lutz , Marina Jecmenica , Özge Ata , Diethard Mattanovich

{"title":"Integration of strain and process optimization to increase autotrophic growth of engineered Komagataella phaffii","authors":"Michael Baumschabl , Lisa Lutz , Marina Jecmenica , Özge Ata , Diethard Mattanovich","doi":"10.1016/j.nbt.2025.11.010","DOIUrl":"10.1016/j.nbt.2025.11.010","url":null,"abstract":"<div><div>Synthetic autotrophs are a promising platform for sustainable bioproduction using CO<sub>2</sub> as substrate. The methylotrophic yeast <em>Komagataella phaffii</em> has been engineered to use CO<sub>2</sub> as the sole carbon source by integration of the Calvin–Benson–Bassham (CBB) cycle, based on its native methanol assimilating xylulose monophosphate (XuMP) cycle. Initial growth rates were low, but could be doubled by adaptive laboratory evolution (ALE). Beneficial mutations led to a decrease of CBB cycle reactions, indicating further limitations. During this study, temperature was identified as one of the key process parameters to improve autotrophic growth. For this reason, a new round of adaptive laboratory evolution was performed at the identified optimal cultivation temperature of 25°C, resulting in clones growing up to 50 % faster compared to the control strain. Whole genome re-resequencing followed by reverse engineering helped to identify first key mutations of the evolved strains. In addition, targeted engineering was performed by increasing the copy number of the key gene of the CBB cycle, RuBisCO, which is the bottleneck for carbon fixation. Combining this strategy together with optimal temperature conditions for cultivation, boosted maximum specific growth rates of the autotrophic <em>K. phaffii</em> strain. In comparison to ALE, the targeted engineering still is lagging behind a bit. Starting from the initial condition, growth was improved more than 2.5-fold in this study reaching a maximum of 0.025 h<sup>−1</sup>.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 110-119"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145636642","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

Generation and NGS profiling of naïve rabbit antibody libraries for efficient monoclonal antibody selection naïve兔抗体库的生成和NGS分析，用于高效的单克隆抗体选择。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-12-11 DOI: 10.1016/j.nbt.2025.12.002

Santiago Rodríguez, Aina García-García, Rosario Martín, Teresa García

{"title":"Generation and NGS profiling of naïve rabbit antibody libraries for efficient monoclonal antibody selection","authors":"Santiago Rodríguez, Aina García-García, Rosario Martín, Teresa García","doi":"10.1016/j.nbt.2025.12.002","DOIUrl":"10.1016/j.nbt.2025.12.002","url":null,"abstract":"<div><div>Rabbit monoclonal antibodies (mAbs) are highly valued for their exceptional affinity and specificity, yet naïve rabbit antibody libraries generated by phage display remain rarely described<strong>,</strong> limiting the in vitro exploitation of the unique rabbit immune repertoire. To address this gap, we constructed two naïve scFv phage display libraries—one incorporating κ and the other λ light chains—assembled from bone marrow, spleen, and peripheral blood of rabbits from two breeds. Together, they yielded a combined theoretical diversity of 7 × 10⁹ independent clones, with the λ library providing access to a rarely explored component of the rabbit repertoire. Next-generation sequencing (NGS) confirmed the high genetic diversity of the libraries and allowed exploration of antibody repertoire regions in rabbits that have been scarcely characterized. A diverse panel of scFvs specific to two major sesame allergens (11S globulins and 2S albumins) have been isolated from both libraries as a proof of concept of their functionality. Selected scFvs were successfully reformatted into full-length rabbit IgG retaining target specificity and expressed in CHO cells at high expression yields. Overall, the constructed libraries provide a robust and versatile resource for antibody discovery, enabling the development of high-quality rabbit mAbs for applications in diagnostics, food safety, and broader analytical applications.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 127-140"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145752193","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

Studying biosorption materials for water PFAS removal in a flow-through configuration 研究生物吸附材料在流动配置中去除水中PFAS。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-11-29 DOI: 10.1016/j.nbt.2025.11.011

Marta Senofonte , Giulia Simonetti , Sara Cerra , Stefano Parisi , Naima Blal , Francesca Buiarelli , Patrizia Di Filippo , Ida Pettiti , Ilaria Fratoddi , Carmela Riccardi , Marco Petrangeli Papini , Laura Lorini

{"title":"Studying biosorption materials for water PFAS removal in a flow-through configuration","authors":"Marta Senofonte , Giulia Simonetti , Sara Cerra , Stefano Parisi , Naima Blal , Francesca Buiarelli , Patrizia Di Filippo , Ida Pettiti , Ilaria Fratoddi , Carmela Riccardi , Marco Petrangeli Papini , Laura Lorini","doi":"10.1016/j.nbt.2025.11.011","DOIUrl":"10.1016/j.nbt.2025.11.011","url":null,"abstract":"<div><div>Per- and polyfluoroalkyl substances (PFAS) are persistent pollutants of global concern due to their strong C–F bonds and exceptional resistance to degradation, which make their removal challenging. Conventional treatment methods such as reverse osmosis, nanofiltration, and activated carbon adsorption often show low efficiency for long-chain PFAS and require high operational costs. This study investigates a sustainable adsorption strategy using pinewood-derived biochar (PW) produced at 850 °C and 1000 °C, with varying particle sizes and including surface functionalization by cetyltrimethylammonium bromide (CTAB). Adsorption experiments were performed in a continuous flow reactor (CSTR) to ensure reproducibility, effective phase separation, and dynamic flow conditions. CTAB-functionalized PW biochar exhibited enhanced adsorption capacity compared to unmodified biochar and comparable performance to commercial activated carbon in long-chain PFAS removal. The use of pinewood residues as a feedstock demonstrates a valorization pathway for industrial and forestry by-products, aligning with circular economy principles and sustainable waste management. Overall, the study highlights engineered biochar as an efficient, low-cost, and environmentally friendly alternative to conventional adsorbents, offering a promising solution for PFAS remediation that integrates pollutant removal with resource recovery.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 100-109"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145648961","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

Exploring protein secretion-modulating and structural features of pro-peptides from S8 class subtilisin-like serine proteases in Bacillus subtilis 探讨枯草芽孢杆菌S8类枯草素样丝氨酸蛋白酶前肽的蛋白分泌调节及结构特征。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-11-10 DOI: 10.1016/j.nbt.2025.11.002

Stefano Grasso , Tobias Schilling , Liselott Schilling , Sven Hutjens , Minia Antelo-Varela , Andreas Otto , Dörte Becher , Tjeerd van Rij , Jan Maarten van Dijl

{"title":"Exploring protein secretion-modulating and structural features of pro-peptides from S8 class subtilisin-like serine proteases in Bacillus subtilis","authors":"Stefano Grasso , Tobias Schilling , Liselott Schilling , Sven Hutjens , Minia Antelo-Varela , Andreas Otto , Dörte Becher , Tjeerd van Rij , Jan Maarten van Dijl","doi":"10.1016/j.nbt.2025.11.002","DOIUrl":"10.1016/j.nbt.2025.11.002","url":null,"abstract":"<div><div>One of the most common post-translational modifications occurring in bacteria is proteolytic cleavage. Particularly, in secretory proteins this cleavage is observed for specific protein segments, named pro-peptides, which are frequently located between the signal peptide and the mature protein. Pro-peptides are known to serve two main functions, namely a chaperone function in protein folding and an enzyme-inhibiting function. To date, the contribution of pro-peptides to protein secretion is not fully understood. Therefore, the present study was aimed at assessing possible roles of the N-terminal pro-peptides from the S8 class subtilisin-like serine proteases Bpr, Vpr and WprA of <em>Bacillus subtilis</em> in protein secretion. To this end, the signal peptides of these proteases were fused to the alkaline phosphatase PhoA of <em>Escherichia coli</em> with or without the cognate pro-peptides. Subsequently, secretion of the PhoA protein and its activity were assessed in the <em>B. subtilis</em> type strain 168 or the genome-reduced strain IIG-Bs27–39 and correlated to the respective pro-peptide structure as predicted by AlphaFold. Altogether, the obtained results suggest that the contributions of the investigated pro-peptides to protein secretion in <em>B. subtilis</em> are related to a combination of their functions in protein folding, their own folding state, and the applied strain.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 7-16"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145505612","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

Evidence for nanoparticle-enhanced ultrasound-based In-Planta transformation and transgenerational inheritance in wheat 基于纳米粒子增强超声的小麦植株内转化和跨代遗传证据。

IF 4.9 2区生物学

New biotechnology Pub Date : 2026-03-25 Epub Date: 2025-12-18 DOI: 10.1016/j.nbt.2025.12.003

Fariba Rafiei , Nirman Nepal , Trever Thurgood , Roland Wilhelm , Steve Scofield , Mohsen Mohammadi

{"title":"Evidence for nanoparticle-enhanced ultrasound-based In-Planta transformation and transgenerational inheritance in wheat","authors":"Fariba Rafiei , Nirman Nepal , Trever Thurgood , Roland Wilhelm , Steve Scofield , Mohsen Mohammadi","doi":"10.1016/j.nbt.2025.12.003","DOIUrl":"10.1016/j.nbt.2025.12.003","url":null,"abstract":"<div><div>Here, we describe a series of evidence that most parsimoniously explains DNA delivery and integration of transgene into wheat genome by using an ultrasound-mediated, nanoparticle-based method without the need for tissue culture. This approach offers a potentially high-throughput, amenable to automation, and a cost-effective strategy for transforming mature wheat seeds and stably transmitting transgenes across generations, with stable inheritance confirmed through the T<sub>3</sub> generation. The method successfully delivered transgenes, via the pCAMBIA1305.1 plasmid, to multiple wheat varieties, including ‘Apogee’, ‘Sonora-64’ and ‘Opata-85’, indicating its potential as a genotype-independent platform for plant genetic engineering. Ultrasound treatment transiently increases the permeability of seed cell walls and membranes, facilitating the uptake of mesoporous silica nanoparticles coated with polyethyleneimine that are loaded with plasmid DNA. Quantitative PCR, segregation analysis, and whole-genome sequencing confirmed variable copy number and stable integration of segments of plasmid DNA across multiple loci. Transgene expression was validated using GUS staining and PCR assays across successive generations, supporting robust transgenerational inheritance. By eliminating the need for callus induction or genotype-specific regeneration protocols, this seed-transformation method offers a scalable, cost-effective, and potentially game-changing approach for the genetic transformation of wheat and other recalcitrant crop species.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"91 ","pages":"Pages 141-145"},"PeriodicalIF":4.9,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145800530","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