Journal of biotechnology最新文献_第5页

Experimental immobilization of Clostridium histolyticum collagenases on green-synthesized CuO nanoparticles supported by bioinformatics analysis 绿色合成氧化铜纳米颗粒固定化溶组织梭菌胶原酶的实验研究。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-29 DOI: 10.1016/j.jbiotec.2025.11.019

Farzaneh Barati , Fakhrisadat Hosseini , Parinaz Ghadam , Seyed Shahriar Arab

Enzymes are widely used in various industries due to their eco-friendly nature, high selectivity, and ability to operate under mild conditions. However, large-scale applications are often hindered by challenges such as limited stability, high costs, and difficulties in recyclability. Enzyme immobilization presents a promising strategy to overcome these limitations. Microbial collagenases, particularly those from Clostridium histolyticum, play a crucial role in industrial and biomedical applications. In silico analyses effectively predict enzyme immobilization outcomes, minimizing experimental time and cost. This study employed enzyme surface analysis, a bioinformatics approach, to determine the surface charge type of the support for the physical immobilization of C. histolyticum collagenases. The analysis identified the most suitable regions on the enzyme surfaces for attachment—positively charged and distant from critical sites—ensuring minimal disruption to activity. Copper oxide (CuO) nanoparticles (NPs) were synthesized using walnut green husk extract and subsequently characterized. The green-synthesized CuO NPs were monodisperse and spherical (∼40 nm) with a negatively charged surface, confirming their suitability as supports for enzyme immobilization. Employing the CuO NPs as supports resulted in an immobilization yield of 42.15 % and an activity yield of 146.2 %. Further optimization of immobilization conditions could improve IY. The high AY suggests that immobilization did not hinder enzyme function but may have enhanced enzyme-substrate affinity and increased local substrate concentration, thereby boosting enzyme efficiency.

酶因其生态友好、高选择性和在温和条件下工作的能力而广泛应用于各个行业。然而，大规模应用经常受到诸如稳定性有限、成本高和可回收性困难等挑战的阻碍。酶固定化是克服这些限制的一种很有前途的策略。微生物胶原酶，特别是来自溶组织梭菌的胶原酶，在工业和生物医学应用中起着至关重要的作用。在硅分析有效地预测酶固定结果，最大限度地减少实验时间和成本。本研究采用酶表面分析这一生物信息学方法，确定了溶组织菌胶原酶物理固定载体的表面电荷类型。分析确定了酶表面最适合附着的区域-带正电且远离关键位点-确保对活性的干扰最小。以核桃绿壳提取物为原料合成了氧化铜纳米颗粒，并对其进行了表征。绿色合成的CuO NPs为单分散球形（~40nm），表面带负电荷，证实了其作为固定化酶载体的适宜性。采用CuO NPs作为载体，固定化产率为42.15%，活性产率为146.2%。进一步优化固定条件可以提高IY。高AY表明固定化不会阻碍酶的功能，但可能增强了酶与底物的亲和力，增加了局部底物浓度，从而提高了酶的效率。

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引用次数: 0

Directional lactic acid production could mitigate antibiotic and ARGs pollution 定向乳酸生产可以减轻抗生素和ARGs的污染

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-27 DOI: 10.1016/j.jbiotec.2025.11.014

Fubin Yin , Tianjing Lian , Haoying Qu , Dongan Cui , Xiaomeng Li , Yangyang Cai , Hongmin Dong

The technology of directional lactic acid production (DLAP) for animal manure is an innovative way of value-added utilization, but the influence of residual antibiotics in animal manure on the DLAP is still unclear, especially the degradation rate and degradation pathways of antibiotic with residues lactic acid bacteria (LAB). In this study, the most common residual antibiotic (Sulfachloropyridazine, SCP) and LAB (Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus casei) were employed as the research objects, the SCP degradation, degradation pathways and antibiotic resistance genes (ARGs) with different LAB were investigated in pure culture. Results showed that LAB has a positive effect on the reduction in ARGs (sul1, sul2, sul3, sulA, intI1 and tnp A). The SCP degradation rates were 90.3 %, 85.1 %, 86.1 % and 84.0 % with the growth of Lactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus casei at 50 mg/L, respectively. And the SCP degradation pathway is different for LAB, but the main decomposition functions, including S-N bond cleavage, CH₃ release, SO₂ release, nitration, hydroxylation, NH₂ release and C-N bond cleavage, are similar. And most of intermediate products of SCP were lower toxicity compared to the parent compound. The findings proved the DLAP process as a guidance for a potential solution for reducing antibiotic and ARGs contamination.

畜禽粪便定向产乳酸（DLAP）技术是一种创新的增值利用方式，但畜禽粪便中残留抗生素对DLAP的影响尚不清楚，尤其是抗生素与残留乳酸菌（LAB）的降解速率和降解途径。本研究以最常见的残留抗生素磺胺氯吡嗪（磺胺氯吡嗪）和乳酸菌（保加利亚乳杆菌、嗜酸乳杆菌、植物乳杆菌和干酪乳杆菌）为研究对象，在纯培养中研究了不同乳酸菌对SCP的降解、降解途径和抗生素抗性基因（ARGs）。结果表明，LAB对ARGs （sul1、sul2、sul3、sulA、intI1和tnp a）的减少具有积极作用。当保加利亚乳杆菌、嗜酸乳杆菌、植物乳杆菌和干酪乳杆菌生长在50 mg/L时，SCP的降解率分别为90.3 %、85.1 %、86.1 %和84.0 %。LAB的SCP降解途径不同，但主要分解功能相似，包括S-N键裂解、CH3释放、SO2释放、硝化、羟基化、NH2释放和C-N键裂解。与母体化合物相比，SCP的大部分中间产物毒性较低。研究结果证明，DLAP工艺为减少抗生素和ARGs污染的潜在解决方案提供了指导。

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引用次数: 0

Sodium alginate-immobilized epoxide hydrolase: A multifaceted strategy for enhanced stability, reusability, and catalytic performance 海藻酸钠固定化环氧化物水解酶：提高稳定性、可重用性和催化性能的多方面策略

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-27 DOI: 10.1016/j.jbiotec.2025.11.017

Zongzhong Yu , Meinan Zhu , Xiao Gu , Zhongkun Wu , Peiqin Chen , Chunying Jin , Junning Zhao , Guangya Zhang , Wei Jiang

Immobilized macromolecular enzymes effectively address critical challenges including environmental sensitivity, thermal instability, and non-reusability, thereby establishing the fundamental and practical significance of enzyme immobilization research. This study employed sodium alginate (SA), ZIF-8, and Bi-EA materials to immobilize Aspergillus carlsbadensis-derived epoxide hydrolase (AcEH). Comparative analysis revealed SA's superior immobilization efficiency. Material characterization via SEM, FTIR, and XRD was performed, followed by optimization of SA immobilization parameters. The immobilized enzyme maintained activity over 6 reuse cycles. Incorporating 0.5 wt% carboxymethyl cellulose (CMC) extended AcEH@SA-CMC reusability to 8 cycles. Magnetic modification through Fe₃O₄ powder incorporation yielded AcEH@SA-CMC- Fe₃O₄ hydrogel with enhanced recoverability. Results demonstrate SA's exceptional biocompatibility underpins its immobilization efficacy, while CMC co-entanglement creates a porous network facilitating mass transfer and mechanical reinforcement. Fe₃O₄ powder plays a promoting role in improving enzyme activity. This methodology not only evaluates immobilization matrices but establishes an optimized protocol for macromolecular AcEH immobilization.

固定化大分子酶有效地解决了环境敏感性、热不稳定性和不可重复使用等关键挑战，从而确立了酶固定化研究的基础和现实意义。本研究采用海藻酸钠（SA）、ZIF-8和Bi-EA材料固定化卡尔斯巴登曲霉衍生环氧化物水解酶（AcEH）。对比分析表明，SA具有较好的固定化效果。通过SEM， FTIR和XRD对材料进行了表征，并对SA的固定参数进行了优化。固定化酶在6次重复使用循环中保持活性。加入0.5 wt%羧甲基纤维素（CMC）将AcEH@SA-CMC可重复使用延长至8次循环。通过掺入Fe3O4粉末进行磁改性，得到了回收率提高的AcEH@SA-CMC- Fe3O4水凝胶。结果表明，SA的特殊生物相容性支撑了其固定效果，而CMC的共缠结形成了多孔网络，促进了传质和机械加固。Fe3O4粉末对提高酶活性有促进作用。该方法不仅评价了固定基质，而且建立了大分子AcEH固定的优化方案。

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引用次数: 0

Human beta-defensin 3-functionalized Fe/GMP nanozyme for multifunctional antimicrobial and anticancer activity against Helicobacter pylori-associated gastrointestinal cancer 人β -防御素3功能化铁/GMP纳米酶对幽门螺杆菌相关胃肠道肿瘤的多功能抗菌和抗癌活性

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-26 DOI: 10.1016/j.jbiotec.2025.11.018

Yoganathan Kamaraj , Veenayohini Kumaresan , Jinhao Hu , Daochen Zhu

Background

The rising incidence of Helicobacter pylori-associated infections and colorectal cancer necessitates multifunctional therapeutics targeting both microbial and tumor pathologies. However, antimicrobial peptides (AMPs) like human β-defensin 3 (hBD-3) faces challenges such as proteolytic degradation and poor bioavailability. To address these limitations, we developed a novel hybrid nanozyme by immobilizing hBD-3 onto a guanosine monophosphate-coordinated iron framework (Fe/GMP), known for its biocompatibility and intrinsic peroxidase-like activity.

Methods

The hBD-3@Fe/GMP nanozyme was synthesized and characterized for its physicochemical and catalytic properties. Its antibacterial activity against H. pylori was assessed via agar diffusion, biofilm inhibition, and virulence gene expression. Anti-colon cancer potential was evaluated in HT-29 cells using assays for cytotoxicity, ROS generation, apoptosis, migration inhibition, and redox biomarkers. in vivo antitumor efficacy was studied using a colon carcinoma xenograft mice model.

Results

hBD-3@Fe/GMP showed enhanced peroxidase-like activity with elevated ROS production than Fe/GMP alone. It significantly inhibited H. pylori growth, biofilm formation, and virulence gene expression (babA, alpA, alpB, flaA, flaB). In HT-29 cells, it induced cytotoxicity, suppressed antioxidant enzymes, and triggered apoptosis via p53 and Bax upregulation and Bcl-2 downregulation. In tumor xenograft model, it reduced tumor volume and weight.

Conclusion

This study reports, for the first time, a hybrid nanozyme that synergistically integrates the antimicrobial potency of hBD-3 with the catalytic redox functionality of Fe/GMP, offering dual action against H. pylori infection and colorectal cancer. The multifunctional therapeutic potential and biocompatibility of hBD-3@Fe/GMP mark a novel paradigm in the development of next-generation nanozymes for combinatorial infectious and oncological interventions.

背景：幽门螺杆菌相关感染和结直肠癌的发病率不断上升，需要针对微生物和肿瘤病理的多功能治疗。然而，抗菌肽（AMPs）如人β-防御素3 （hBD-3）面临着蛋白水解降解和生物利用度差等挑战。为了解决这些限制，我们通过将hBD-3固定在鸟苷单磷酸配位铁框架（Fe/GMP）上开发了一种新型杂交纳米酶，该框架以其生物相容性和内在过氧化物酶样活性而闻名。方法合成hBD-3@Fe/GMP纳米酶，并对其理化性质和催化性能进行表征。通过琼脂扩散、生物膜抑制和毒力基因表达评价其对幽门螺杆菌的抑菌活性。通过细胞毒性、ROS生成、凋亡、迁移抑制和氧化还原生物标志物的测定来评估HT-29细胞的抗结肠癌潜力。采用结肠癌异种移植小鼠模型研究其体内抗肿瘤效果。ResultshBD-3@Fe/GMP显示，与单独Fe/GMP相比，过氧化物酶样活性增强，ROS生成增加。显著抑制幽门螺杆菌生长、生物膜形成和毒力基因（babA、alpA、alpB、flaA、flaB）表达。在HT-29细胞中，它通过上调p53和Bax，下调Bcl-2，诱导细胞毒性，抑制抗氧化酶，引发细胞凋亡。在异种肿瘤移植模型中，它能减小肿瘤体积和重量。结论本研究首次报道了一种将hBD-3的抗菌效力与Fe/GMP的催化氧化还原功能协同结合的杂交纳米酶，具有抗幽门螺杆菌感染和结直肠癌的双重作用。hBD-3@Fe/GMP的多功能治疗潜力和生物相容性标志着用于联合感染和肿瘤干预的下一代纳米酶的开发的新范式。

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引用次数: 0

Emerging strategies for enhanced glutathione biosynthesis and its biomedical applications 加强谷胱甘肽生物合成及其生物医学应用的新策略

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-25 DOI: 10.1016/j.jbiotec.2025.11.016

Yajie Wang , Ruixue Ma , Jia Zhou , Zhongbiao Tan , Hongliang Li , Hao Shi , Yegui Zhang

Glutathione (GSH), a ubiquitous bioactive tripeptide, plays a central role in antioxidant defense, detoxification, and cellular metabolism. This review highlights advanced strategies to enhance GSH production, emphasizing strain improvement via traditional mutagenesis and genetic engineering. Optimization of microbial fermentation through carbon source selection, amino acid precursors, and ATP regeneration combined with metabolic engineering approaches, has significantly improved GSH yields in Saccharomyces cerevisiae and Escherichia coli. Clinically, GSH is used in managing liver diseases, oxidative stress, and immune dysfunction. Emerging applications, particularly in oncology, show promise. This review also compares production methods such as chemical synthesis, enzymatic catalysis, and microbial fermentation, highlighting microbial fermentation for its sustainability, cost-efficiency, and scalability. Future research should focus on refining detection methods, elucidating GSH’s therapeutic mechanisms, and broadening its applications across pharmaceuticals, nutrition, and cosmetics.

谷胱甘肽（GSH）是一种普遍存在的生物活性三肽，在抗氧化防御、解毒和细胞代谢中起着核心作用。本文综述了提高谷胱甘肽产量的先进策略，强调了通过传统的诱变和基因工程对菌株进行改良。通过碳源选择、氨基酸前体和ATP再生结合代谢工程方法优化微生物发酵，显著提高了酿酒酵母和大肠杆菌的谷胱甘肽产量。在临床上，谷胱甘肽被用于治疗肝脏疾病、氧化应激和免疫功能障碍。新兴的应用，特别是在肿瘤学领域，显示出了希望。本文还比较了化学合成、酶催化和微生物发酵等生产方法，重点介绍了微生物发酵的可持续性、成本效益和可扩展性。未来的研究应集中于改进检测方法，阐明谷胱甘肽的治疗机制，并扩大其在制药、营养和化妆品领域的应用。

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引用次数: 0

Methanol-induced metabolic disturbances in protein expression in Pichia pastoris: A review of the interaction between amino acid synthesis and higher alcohol pathways 毕赤酵母中甲醇诱导的蛋白质表达代谢紊乱：氨基酸合成与高级酒精途径相互作用的综述

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-25 DOI: 10.1016/j.jbiotec.2025.11.015

Lifen He , Danling Tang , Fan Feng , Tao Hong , Mingjing Zheng , Xiping Du , Yanbing Zhu , Yuanfan Yang , Zedong Jiang , Fan He , Lijun Li , Zhipeng Li , Hui Ni , Qingbiao Li

Pichia pastoris is a prominent host for recombinant protein production, utilizing the methanol-inducible Alcohol Oxidase 1 (AOX1) promoter to express thousands of heterologous proteins. However, methanol induction, while driving high-level protein expression, simultaneously imposes a pronounced metabolic burden. This review highlights that a balanced and ample supply of amino acids is indispensable for efficient protein synthesis. The induction process leads to the accumulation of by-products, notably higher alcohols, which can negatively impact protein quality. The dynamic interplay between these two pathways is not fully understood. In addition, the lack of quantitative models for their competition over shared precursors hinders rational metabolic engineering and impedes the industrial exploitation of P. pastoris. Therefore, this review systematically dissects the link between methanol induction, amino acid metabolism, and higher alcohol synthesis. It specifically examines how methanol-triggered perturbations in amino acid metabolism propagate to modulate higher alcohol formation. Special attention is given to key metabolic nodes, such as pyruvate and α-ketoisovalerate, where carbon flux diverges between biosynthesis and by-product formation. This review provides significant implications for a deeper understanding of the metabolic processes in P. pastoris, offering comprehensive mechanistic insights and guidance for optimizing yeast fermentation and enhancing recombinant protein production.

毕赤酵母是重组蛋白生产的重要宿主，利用甲醇诱导的乙醇氧化酶1 （AOX1）启动子表达数千种异源蛋白。然而，甲醇诱导在驱动高水平蛋白表达的同时，也会带来明显的代谢负担。这篇综述强调了平衡和充足的氨基酸供应对于有效的蛋白质合成是必不可少的。诱导过程导致副产物的积累，特别是高醇，这可能对蛋白质质量产生负面影响。这两种途径之间的动态相互作用尚不完全清楚。此外，缺乏对共享前体竞争的定量模型，阻碍了合理的代谢工程，阻碍了巴氏酵母的工业开发。因此，本文系统地剖析了甲醇诱导、氨基酸代谢和高级醇合成之间的联系。它特别研究了甲醇触发的氨基酸代谢扰动如何传播以调节更高的酒精形成。特别关注关键的代谢节点，如丙酮酸和α-酮异戊酸，其中碳通量在生物合成和副产物形成之间偏离。这一综述为深入了解酵母的代谢过程提供了重要的意义，为优化酵母发酵和提高重组蛋白的生产提供了全面的机制见解和指导。

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引用次数: 0

Production of high-nutrient protein sources by pre-secondary solid-state fermentation of functional microbiota (Pre-Sec-SSF-FM) from dehulled chinese distillers grains 功能菌群（Pre-Sec-SSF-FM）预二级固态发酵生产高营养蛋白源的研究

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-21 DOI: 10.1016/j.jbiotec.2025.11.012

Lian Zhang , Xiaojuan Shen , Zhi Wang , Yuansong Peng , Jianping Yang , Yafan Cai , Shilei Wang , Jingliang Xu , Hanjie Ying

Chinese distillers grains (CDGs) are used as low-cost feed additives, because they have issues such as high crude fiber content (> 24 %), low crude protein content (< 17 %), and low digestibility (< 16 %). This study aimed to evaluate whether pre-secondary solid-state fermentation using functional microbiota can improve protein digestibility and reduce fiber content in CDGs. Results indicated that compared to CDGs, both true and crude protein content in solid-state fermented DCDGs (SSF-DCDGs) showed an upward trend, reaching increases of over 24 %. Compared to DCDGs, crude fiber content decreased; dry matter and protein digestibility increased by 29.54 % and 54.57 % in SSF-DCDGs. Amino acid content rose by 14.92 %, with glycine and alanine increased by 15.36 % and 21.25 % respectively—both being important aromatic flavor compounds. Therefore, the Pre-Sec-SSF-FM technical route can convert low-cost CDGs into a high-nutritional protein sources, offering a novel approach for protein raw material sourcing.

由于中国白酒糟的粗纤维含量高（约为24%），粗蛋白质含量低（< 17%），消化率低（< 16%），因此被用作低成本的饲料添加剂。本研究旨在评估利用功能菌群进行预二级固态发酵是否能提高CDGs的蛋白质消化率和降低纤维含量。结果表明，与CDGs相比，固态发酵DCDGs （SSF-DCDGs）的真蛋白质含量和粗蛋白质含量均呈上升趋势，增幅均在24%以上。与DCDGs相比，粗纤维含量降低；SSF-DCDGs的干物质和蛋白质消化率分别提高29.54%和54.57%。氨基酸含量提高了14.92%，甘氨酸和丙氨酸含量分别提高了15.36%和21.25%，均为重要的芳香风味化合物。因此，Pre-Sec-SSF-FM技术路线可以将低成本的CDGs转化为高营养的蛋白质来源，为蛋白质原料的采购提供了一种新的途径。

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引用次数: 0

Optical, structural, and antifungal properties of nanosilver borate bioactive glass synthesized using gamma rays on the survival of Candida albicans and Candida tropicalis 利用伽马射线合成的纳米硼酸银生物活性玻璃对白色念珠菌和热带念珠菌生存的光学、结构和抗真菌特性。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-21 DOI: 10.1016/j.jbiotec.2025.11.013

E.M. Abou Hussein , Rasha Mohammad Fathy

A New glass system with a composition of; 50.3 B₂O₃+ 21 CaO + 8 MgO + 8 K₂O + 8 Na₂O + 4 P₂O₅ + 0.7 Nano Ag₂O, was prepared by the traditional melting technique. Some of its structural, optical, and physical characteristics were investigated regarding the effect of gamma radiation. The XRD pattern revealed the amorphous nature of the fabricated glass sample. UV–visible absorption spectra revealed an obvious peak before 400 nm due to the absorption of Ag^2 + ions, and a photo-reduction reaction to Ag⁰ was expected due to the impact of gamma irradiation. FTIR absorption spectra displayed structural bands in the region 800–1600 cm⁻¹ recognized to the specific vibrations of BO₃, BO₄ and PO₂. Results displayed optical and structural stability against gamma irradiation doses of 25 and 50 kGy, and revealed a positive effect of irradiation to enhance the antifungal efficiency of the glass or in sterilization processes before usage. The antifungal activity of borate bioglass doped with silver nanoparticles (BG/Ag) against Candida albicans and Candida tropicalis was assessed. BG/Ag was mostly effective against C. albicans. Furthermore, at a dosage of 40.0 mg/mL, BG/Ag considerably reduced biofilm formation of C. albicans and C. tropicalis (90.79 and 75.07 %). At 40 mg/mL, BG/Ag significantly reduced Candida viability, colony counts, and phospholipase activity. BG/Ag can be successfully applied as an attractive biomaterial for the medication of serious fungal infections. The novelty in the study is the synthesis of borate bioactive glass containing nanosilver- and the use of gamma irradiation to improve its biological properties.

一种新型玻璃体系，其组成为；采用传统熔融法制备了50.3 B2O3+ 21 CaO + 8 MgO + 8 K2O + 8 Na2O+ 4 P2O5 + 0.7纳米Ag2O。在伽玛辐射的影响下，研究了它的一些结构、光学和物理特性。XRD图显示了制备的玻璃样品的非晶态性质。由于Ag2+离子的吸收，紫外可见吸收光谱在400nm前出现明显的峰值，并且由于伽马辐照的影响，预计会发生Ag0的光还原反应。FTIR吸收光谱显示出800-1600cm-1区域的结构带，这是BO3、BO4和PO2特有的振动特征。结果表明，在25和50 kGy的伽马辐射剂量下，玻璃具有光学稳定性和结构稳定性，并显示辐照对提高玻璃的抗真菌效率或在使用前的灭菌过程中具有积极作用。研究了纳米银掺杂硼酸盐生物玻璃对白色念珠菌和热带念珠菌的抑菌活性。BG/Ag对白色念珠菌最有效。此外，在40.0mg/mL的剂量下，BG/Ag显著减少了白色念珠菌和热带念珠菌的生物膜形成（90.79%和75.07%）。在40mg/mL浓度下，BG/Ag显著降低念珠菌活力、菌落计数和磷脂酶活性。BG/Ag可以作为一种有吸引力的生物材料成功应用于严重真菌感染的治疗。这项研究的新颖之处在于合成了含有纳米银的硼酸盐生物活性玻璃，并使用伽马射线照射来改善其生物特性。

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引用次数: 0

Halotolerant black yeast Neophaeotheca triangularis as a source of melanin 耐盐黑酵母菌，作为黑色素的来源。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-12 DOI: 10.1016/j.jbiotec.2025.11.011

Mercedes Peña , Patricia Gómez-Villegas , Jose Prados , Consolación Melguizo , Carla C.C.R. de Carvalho

Archaea and bacteria are the most studied extremophiles, but fungi also demonstrate remarkable tolerance, particularly in hypersaline environments such as solar salterns. Among salt-adapted fungi, black yeasts have been shown to be adapted to such environments, having developed defense mechanisms such as the production of melanin, a pigment that plays a crucial role in environmental stress protection. Melanin is a complex, high-molecular-weight polymer widely found across biological kingdoms. During the isolation of microorganisms from samples collected in crystallization ponds in a saltern, a black yeast was found able to produce significant amounts of melanin. The yeast was identified as belonging to the species Neophaeotheca triangularis. The aim of this study was to optimize the cultivation conditions of the strain, to characterize the produced melanin, and to assess its biological activities, including its antitumor and antioxidant properties.

古细菌和细菌是研究最多的极端微生物，但真菌也表现出非凡的耐受性，特别是在高盐环境中，如太阳盐沼。在适应盐的真菌中，黑酵母已经被证明能够适应这样的环境，已经发展出防御机制，如产生黑色素，一种在环境应激保护中起关键作用的色素。黑色素是一种复杂的高分子量聚合物，广泛存在于生物王国中。在从盐沼结晶池中收集的样品中分离微生物时，发现一种黑酵母能够产生大量的黑色素。经鉴定，该酵母菌属Neophaeotheca triangularis。本研究的目的是优化菌株的培养条件，表征所产生的黑色素，并评估其生物活性，包括抗肿瘤和抗氧化性能。

引用次数: 0

FRET two-hybrid assay-based target drug screening in living cells 基于FRET双杂交分析的活细胞靶标药物筛选。

IF 3.9 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology

Pub Date : 2025-11-12 DOI: 10.1016/j.jbiotec.2025.11.010

Beini Sun , Gengqiang Cao , Tian Gan , Yue Wang , Rumeng Qu , Lin Hu , Yuan Chang , Min Hu , Xiaoping Wang , Tongsheng Chen

Precise targeted drug screening is the key to improve the efficiency of tumour targeted therapy. This report presents a live cell FRET two-hybrid assay-based targeted drug screening method (FRET-HBTDS). In FRET-HBTDS, the cells co-expressing donor- and acceptor-labelled targets were cultured in 96-well plates, quantitative FRET imaging was then performed on a self-built automated FRET microscope (FRETscope) well-by-well, and FRET two-hybrid assay was used to obtain the maximum donor-centre FRET efficiency (E_Dmax), maximum acceptor-centre FRET efficiency (E_Amax) and stoichiometric ratios (N_A/N_D). The FRET-HBTDS method was performed on the FRETscope with a 20 × objective for the cells co-expressing CFP-Bcl-xL and YFP-Bak to assess the action of eight compounds (A1331852, S63845, AC, DSF/Cu, Met, REGO, SOFA, ABT199) on the interaction between Bcl-xL and Bak. After 7 h of treatment with these compounds respectively, only the A1331852 group showed significantly lower E_Dmax and E_Amax compared to the control group, and a significant increase in N_A/N_D, suggesting that A1331852 unlocks the direct interaction between Bcl-xL and Bak, thus releasing Bak to induce cell death. In addition, the N_A/N_D of the DSF/Cu group was significantly higher than of the control group, suggesting that DSF/Cu altered the stoichiometry of the Bcl-xL-Bak complex. Our data firmly demonstrate that A1331852 unlocks the binding state of Bcl-xL and Bak, while DSF/Cu modifies the structure of the Bcl-xL-Bak complex. These findings demonstrate that FRET-HBTDS can be used to assess the efficacy of a drug by revealing the binding state and complex molecular structure of the target proteins using FRET technology in living cells, which may be a potential targeted drug screening method.

精确的靶向药物筛选是提高肿瘤靶向治疗效率的关键。本报告提出了一种基于活细胞FRET双杂交测定的靶向药物筛选方法（FRET- hbtds）。在FRET- hbtds中，将共表达供体和受体标记靶标的细胞培养在96孔板中，然后在自建的自动FRET显微镜（FRETscope）上逐孔进行定量FRET成像，并使用FRET双杂交法获得最大供体中心FRET效率（EDmax），最大受体中心FRET效率（EAmax）和化学计量比（NA/ND）。在FRETscope上对共表达CFP-Bcl-xL和YFP-Bak的细胞采用FRET-HBTDS法，以20倍的倍率评估8个化合物（A1331852、S63845、AC、DSF/Cu、Met、REGO、SOFA、ABT199）对Bcl-xL和Bak相互作用的影响。分别用这些化合物处理7h后，只有A1331852组EDmax和EAmax较对照组显著降低，NA/ND显著升高，提示A1331852开启了Bcl-xL与Bak的直接相互作用，从而释放Bak诱导细胞死亡。此外，DSF/Cu组的NA/ND显著高于对照组，表明DSF/Cu改变了Bcl-xL-Bak复合物的化学计量。我们的数据有力地证明了A1331852解锁了Bcl-xL和Bak的结合状态，而DSF/Cu修饰了Bcl-xL-Bak复合物的结构。这些发现表明，FRET- hbtds可以利用FRET技术在活细胞中揭示靶蛋白的结合状态和复杂的分子结构，从而评估药物的疗效，这可能是一种潜在的靶向药物筛选方法。

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