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Extraction of cellulose from halophytic plants for the synthesis of a novel biocomposite 从盐生植物中提取纤维素,用于合成新型生物复合材料。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-15 DOI: 10.1002/bip.23586
Uroosa Ejaz, Yusra Shafquat, Muhammad Sohail, Aizaz Ahmed Shaikh, Muhammad Daniyal Arain, Tehmees Ahmed, Abdullah K. Alanazi

Cellulose nanofibers, a sustainable and promising material with widespread applications, exhibit appreciable strength and excellent mechanical and physicochemical properties. The preparation of cellulosic nanofibers from food or agricultural residue is not sustainable. Therefore, this study was designed to use three halophytic plants (Cressa cretica, Phragmites karka, and Suaeda fruticosa) to extract cellulose for the subsequent conversion to cellulosic nanofibers composites. The other extracted biomass components including lignin, hemicellulose, and pectin were also utilized to obtain industrially valuable enzymes. The maximum pectinase (31.56 IU mL−1), xylanase (35.21 IU mL−1), and laccase (15.89 IU mL−1) were produced after the fermentation of extracted pectin, hemicellulose, and lignin from S. fruticosa, P. karka, and C. cretica, respectively. Cellulose was methylated (with a degree of substitution of 2.4) and subsequently converted into a composite using polyvinyl alcohol. Scanning electron microscopy and Fourier-transform infrared spectroscopy confirmed the successful synthesis of the composites. The composites made up of cellulose from C. cretica and S. fruticosa had a high tensile strength (21.5 and 15.2 MPa) and low biodegradability (47.58% and 44.56%, respectively) after dumping for 3 months in soil, as compared with the composite from P. karka (98.79% biodegradability and 4.9 MPa tensile strength). Moreover, all the composites exhibited antibacterial activity against gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) and gram-positive bacteria (Staphylococcus aureus). Hence, this study emphasizes the possibility for various industrial applications of biomass from halophytic plants.

纤维素纳米纤维是一种具有广泛应用前景的可持续材料,具有可观的强度和优异的机械和物理化学特性。从食物或农业残留物中制备纤维素纳米纤维不是可持续的。因此,本研究旨在利用三种卤叶植物(Cressa cretica、Phragmites karka 和 Suaeda fruticosa)提取纤维素,然后将其转化为纤维素纳米纤维复合材料。提取的其他生物质成分(包括木质素、半纤维素和果胶)也被用来获得有工业价值的酶。从 S. fruticosa、P. karka 和 C. cretica 提取的果胶、半纤维素和木质素经发酵后,分别产生了最大的果胶酶(31.56 IU mL-1)、木聚糖酶(35.21 IU mL-1)和漆酶(15.89 IU mL-1)。纤维素被甲基化(取代度为 2.4),然后用聚乙烯醇转化成复合材料。扫描电子显微镜和傅立叶变换红外光谱证实了复合材料的成功合成。由 C. cretica 和 S. fruticosa 纤维素制成的复合材料在土壤中倾倒 3 个月后具有较高的拉伸强度(21.5 和 15.2 兆帕)和较低的生物降解性(分别为 47.58% 和 44.56%),而 P. karka 的复合材料则具有 98.79% 的生物降解性和 4.9 兆帕的拉伸强度。此外,所有复合材料都对革兰氏阴性菌(大肠杆菌和肺炎克雷伯氏菌)和革兰氏阳性菌(金黄色葡萄球菌)具有抗菌活性。因此,这项研究强调了卤叶植物生物质在各种工业应用中的可能性。
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引用次数: 0
A facile and scalable method to synthesize PEGylated PDMAEMA for gene delivery 用于基因递送的 PEG 化 PDMAEMA 的简便、可扩展合成方法
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-02 DOI: 10.1002/bip.23584
Jie Dou, Shupei Yu, Yuanwei Zhang

In recent years, cationic polymer vectors have been viewed as a promising method for delivering nucleic acids. With the advancement of synthetic polymer chemistry, we can control chemical structures and properties to enhance the efficacy of gene delivery. Herein, a facile, cost-effective, and scalable method was developed to synthesize PEGylated PDMAEMA polymers (PEO-PDMAEMA-PEO), where PEGylation could enable prolonged polyplexes circulation time in the blood stream. Two polymers of different molecular weights were synthesized, and polymer/eGFP polyplexes were prepared and characterized. The correlation between polymers' molecular weight and physicochemical properties (size and zeta potential) of polyplexes was investigated. Lipofectamine 2000, a commercial non-viral transfection reagent, was used as a standard control. PEO-PDMAEMA-PEO with higher molecular weight exhibited slightly better transfection efficiency than Lipofectamine 2000, and the cytotoxicity study proved that it could function as a safe gene vector. We believe that PEO-PDMAEMA-PEO could serve as a model to investigate more potential in the gene delivery area.

近年来,阳离子聚合物载体被视为一种很有前景的核酸递送方法。随着合成聚合物化学的发展,我们可以通过控制化学结构和性质来提高基因递送的效果。本文开发了一种简便、经济、可扩展的方法来合成 PEG 化 PDMAEMA 聚合物(PEO-PDMAEMA-PEO),其中 PEG 化可以延长多聚体在血流中的循环时间。我们合成了两种不同分子量的聚合物,并制备和表征了聚合物/eGFP 多聚体。研究了聚合物分子量与多聚体理化性质(大小和 zeta 电位)之间的相关性。以商用非病毒转染试剂 Lipofectamine 2000 作为标准对照。分子量更高的 PEO-PDMAEMA-PEO 的转染效率略高于 Lipofectamine 2000,细胞毒性研究证明它可以作为一种安全的基因载体。我们相信,PEO-PDMAEMA-PEO 可以作为一个模型,在基因递送领域发挥更大的潜力。
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引用次数: 0
Synthesis of PEGylated amphiphilic block copolymers with pendant linoleic moieties by combining ring-opening polymerization and click chemistry 结合开环聚合和点击化学,合成具有亚油酸悬垂分子的 PEG 化两亲嵌段共聚物
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-29 DOI: 10.1002/bip.23582
Julian D. Porras, Ivonne L. Diaz, Leon D. Perez

This study focused on synthesizing and characterizing PEGylated amphiphilic block copolymers with pendant linoleic acid (Lin) moieties as an alternative to enhance their potential in drug delivery applications. The synthesis involved a two-step process, starting with ring-opening polymerization of ε-caprolactone (CL) and propargylated cyclic carbonate (MCP) to obtain PEG-b-P(CL-co-MCP) copolymers, which were subsequently modified via click chemistry. Various reaction conditions were explored to improve the yield and efficiency of the click chemistry step. The use of anisole as a solvent, N-(3-azidopropyl)linoleamide as a substrate, and a reaction temperature of 60°C proved to be highly efficient, achieving nearly 100% conversion at a low catalyst concentration. The resulting copolymers exhibited controlled molecular weights and low polydispersity, confirming the successful synthesis. Furthermore, click chemistry allows for the attachment of Lin moieties to the copolymer, enhancing its hydrophobic character, as deduced from their significantly lower critical micelle concentration than that of traditional PEG-b-PCL systems, which is indicative of enhanced stability against dilution. The modified copolymers exhibited improved thermal stability, making them suitable for applications that require high processing temperatures. Dynamic light scattering and transmission electron microscopy confirmed the formation of micellar structures with sizes below 100 nm and minimal aggregate formation. Additionally, 1H NMR spectroscopy in deuterated water revealed the presence of core-shell micelles, which provided higher kinetic stability against dilution.

本研究的重点是合成和表征带有亚油酸(Lin)悬垂分子的 PEG 化两亲嵌段共聚物,以提高其在药物递送应用中的潜力。合成过程分为两步,首先对ε-己内酯(CL)和丙炔化环碳酸酯(MCP)进行开环聚合,得到 PEG-b-P(CL-co-MCP)共聚物,然后通过点击化学进行改性。研究人员探索了各种反应条件,以提高点击化学步骤的产率和效率。事实证明,使用苯甲醚作为溶剂、N-(3-叠氮丙基)亚油酰胺作为底物以及 60°C 的反应温度具有很高的效率,在催化剂浓度较低的情况下,转化率接近 100%。生成的共聚物分子量可控,多分散性低,证明合成成功。此外,点击化学还可将 Lin 分子附着到共聚物上,从而增强其疏水特性,这一点可从其临界胶束浓度明显低于传统的 PEG-b-PCL 系统推断出来,这表明其稀释稳定性得到了增强。改性共聚物具有更高的热稳定性,因此适用于需要较高加工温度的应用领域。动态光散射和透射电子显微镜证实了胶束结构的形成,其尺寸低于 100 nm,且聚合体形成极少。此外,氘化水中的 1H NMR 光谱显示存在核壳胶束,这种胶束在稀释时具有更高的动力学稳定性。
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引用次数: 0
Sustainable strategies towards better utilization of synthetic polymers 更好地利用合成聚合物的可持续战略。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-26 DOI: 10.1002/bip.23581
G. Shanthi, J. Beula Isabel, Rosbin Thankachan, M. Premalatha

The abstract provides an overview of a study focused on analyzing diverse strategies to achieve sustainable utilization of synthetic polymers through effective waste management. The escalating global consumption of synthetic polymers has precipitated a concerning increase in plastic waste and environmental degradation. To address this challenge, novel materials with specified application goals, such as engineered plastic, have been developed and are intended for recycling and reuse. Despite the reuse and recycling, when plastic gets disposed into the environment, the degradation properties of plastics render a direct disposal hazard, posing a significant environmental threat. To mitigate these issues, the concept of replacing specific monomers of engineered synthetic plastics with bio-alternatives or blending them with other polymers to enhance sustainability and environmental compatibility has emerged. In this study, Acrylonitrile Butadiene Styrene (ABS) plastic is the focal material, and three distinct investigations were conducted. First, replacing ABS plastic's butadiene monomer with natural rubber was explored for its properties and environmental impact. Second, ABS plastic was blended with virgin, recycled, and bio-alternatives of PET (polyethylene terephthalate) and PVC (polyvinyl chloride) polymers. Lastly, recycled ABS blended with recycled PET and PVC was analyzed for mechanical properties. Comparative assessments of these blends were made based on mechanical properties, carbon emissions, and cost-effectiveness. The study determined that the r-ABS/r-PVC (recycled) blend exhibited the most favorable characteristics for practical application.

摘要概述了一项研究,其重点是分析通过有效的废物管理实现合成聚合物可持续利用的各种战略。随着全球合成聚合物消费量的不断攀升,塑料废弃物的增加和环境退化令人担忧。为了应对这一挑战,人们开发出了具有特定应用目标的新型材料,如工程塑料,并打算进行回收和再利用。尽管进行了再利用和再循环,但当塑料被丢弃到环境中时,塑料的降解特性会造成直接的丢弃危险,对环境构成严重威胁。为了缓解这些问题,出现了用生物替代品替代工程合成塑料的特定单体或将其与其他聚合物混合以提高可持续性和环境兼容性的概念。本研究以丙烯腈-丁二烯-苯乙烯(ABS)塑料为重点材料,进行了三项不同的研究。首先,研究了用天然橡胶替代 ABS 塑料的丁二烯单体的特性和对环境的影响。其次,将 ABS 塑料与 PET(聚对苯二甲酸乙二酯)和 PVC(聚氯乙烯)聚合物的原生、回收和生物替代品混合。最后,分析了回收 ABS 与回收 PET 和 PVC 混合后的机械性能。根据机械性能、碳排放量和成本效益对这些混合物进行了比较评估。研究结果表明,r-ABS/r-PVC(回收)混合物在实际应用中表现出最有利的特性。
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引用次数: 0
Hydroxyapatite-loaded macroporous calcium alginate hydrogels: Preparation, characterization, and in vitro evaluation 负载羟基磷灰石的大孔海藻酸钙水凝胶:制备、表征和体外评估。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-25 DOI: 10.1002/bip.23583
Maria Drozdova, Alika Makhonina, Daria Gladkikh, Alexander Artyukhov, Leonid Bryukhanov, Yaroslav Mezhuev, Vladimir Lozinsky, Elena Markvicheva

Hydrogels from natural polysaccharides are of great interest for tissue engineering. This study aims (1) to prepare hydroxyapatite-loaded macroporous calcium alginate hydrogels by novel one-step technique using internal gelation in water-frozen solutions; (2) to evaluate their physicochemical properties; (3) to estimate their ability to support cell growth and proliferation in vitro. The structure of the hydrogel samples in a swollen state was studied by confocal laser scanning microscopy and was shown to represent a system of interconnected macropores with sizes of tens micron. The swelling behavior of the hydrogels, their mechanical properties (Young's moduli) in function of a hydroxyapatite content (5–30 mass%) were studied. All hydrogel samples loaded with hydroxyapatite were found to support growth and proliferation of mouse fibroblasts (L929) at long-term cultivation for 7 days. The obtained macroporous composite Ca-Alg-HA hydrogels could be promising for tissue engineering.

天然多糖水凝胶对组织工程具有重要意义。本研究旨在:(1)采用新型一步法技术,在水冷冻溶液中进行内凝胶化,制备负载羟基磷灰石的大孔海藻酸钙水凝胶;(2)评估其理化性质;(3)评估其体外支持细胞生长和增殖的能力。通过激光共聚焦扫描显微镜研究了水凝胶样品在溶胀状态下的结构,结果表明它代表了一个由大小为几十微米的相互连接的大孔组成的系统。研究了水凝胶的溶胀行为及其机械性能(杨氏模量)与羟基磷灰石含量(5-30%)的函数关系。研究发现,所有负载羟基磷灰石的水凝胶样品都能支持小鼠成纤维细胞(L929)在 7 天的长期培养过程中生长和增殖。所获得的大孔复合 Ca-Alg-HA 水凝胶有望用于组织工程。
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引用次数: 0
State diagrams of green peas (Pisum sativum L.) powders with different maltodextrin additions 添加不同麦芽糊精的青豌豆(Pisum sativum L.)粉末的状态图
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-15 DOI: 10.1002/bip.23580
Anna Stępień, Mariusz Witczak

The purpose of this study was to examine the effect of maltodextrin addition on the physical stability of powdered green peas. The evaluation of the physical state of the material was based on the equilibrium water content of the monolayer (Xm) and the glass transition temperatures of the powders at room temperature (Tg) and in the frozen state (Tg'). Graphical sorption characteristic at 25°C was determined using static—gravimetric method while capacity of the monolayer values was calculated from the mathematical GAB model. Differential scanning calorimetry was carried out in order to determine glass transition lines and freezing curves which combine together were used to plot state diagrams. Relationship between Tg and solid content were shown by using Gordon–Taylor model. Freezing data were modeled employing the Clausius–Clapeyron equation and its development—Chen model. Sorption isotherms showed sigmoidal shape characteristic for high-molecular weight materials. Monolayer moisture content varied between 0.047 and 0.106 g water/g solids. The glass transition temperature of anhydrous green peas increased in from 89.9 to 175.6°C while Tg′ value changed from −43.4 to −26.6°C to as a result of 75% polysaccharide addition. The ultimate maximum-freeze-concentration conditions of the powders were observed in range from 0.783 to 0.814 g solids/g sample. Monolayer capacity, Tg and Tg′ values increased with increasing maltodextrin amount in the sample which indicates that the addition of starch hydrolysate has a beneficial effect on the stability of powders stored frozen and at room temperature.

本研究的目的是考察添加麦芽糊精对青豌豆粉物理稳定性的影响。材料物理状态的评估基于单层平衡含水量(Xm)以及粉末在室温(Tg)和冷冻状态(Tg')下的玻璃化转变温度。25°C 时的图形吸附特性是用静态重力法测定的,而单层的容量值则是根据数学 GAB 模型计算出来的。为了确定玻璃化转变线和冻结曲线,采用了差示扫描量热法。使用 Gordon-Taylor 模型显示了 Tg 与固体含量之间的关系。凝固数据采用克劳修斯-克拉皮隆方程及其发展-陈模型。吸附等温线显示出高分子量材料的曲线特征。单层含水量在 0.047 至 0.106 克水/克固体之间变化。由于添加了 75% 的多糖,无水青豌豆的玻璃化温度从 89.9°C 上升到 175.6°C,而 Tg′ 值则从 -43.4°C 变为 -26.6°C。粉末的最终最大冷冻浓度范围为 0.783 至 0.814 克固体/克样品。单层容量、Tg 和 Tg′ 值随着样品中麦芽糊精含量的增加而增加,这表明淀粉水解物的添加对粉末在冷冻和室温下储存的稳定性有好处。
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引用次数: 0
Antibacterial wound dressing with cross-linked electrospun surface from reduced graphene oxide doped polyvinyl alcohol/sodium caseinate blends 掺杂还原氧化石墨烯的聚乙烯醇/酪蛋白酸钠混合物交联电纺表面的抗菌伤口敷料
IF 2.9 4区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-04-05 DOI: 10.1002/bip.23579
Sema Samatya Yilmaz, Ayse Aytac

In this study, a new biomaterial with polyvinyl alcohol (PVA)/sodium caseinate (SodCa)/reduced graphene oxide (rGO) structure was developed. Antibacterial effective nanofibers were successfully produced by electrospinning method from 1%, 3%, 5%, and 7% rGO added PVA/SodCa (60:40, w:w) solution mixtures prepared for use as modern wound dressings. To create a usage area, especially in exuding wounds, hydrophilic PVA/SodCa/rGO electrospun mats were cross-linked by dipping them in a glutaraldehyde (GLA) bath. The surface micrographs of all nanofibers were homogeneous and smooth. rGO-doped biomaterials were obtained as thin nanofibers in the range of 301–348 nm. Nanofibers, which were completely soluble in water, after cross-linking preserved their existence in the range of 87%–81% at the end of the 24th hour in distilled water. It was reported that these biomaterials that persist in an aqueous environment show swelling behavior in the range of 275%–608%. The porosity of uncross-linked pure PVA/SodCa nanofibers increased by 46.75% after cross-linking. Moreover, the tensile strength of cross-linked PVA/SodCa electrospun mats increased in the presence of rGO. Provided that wound dressing is done every 24 h with 3% rGO-doped PVA/SodCa nanofiber and provided that wound dressing is done every 48 h with 5% rGO-doped PVA/SodCa nanofiber showed antibacterial activity against S. aureus as 99.38% and 99.55%, respectively.

本研究开发了一种具有聚乙烯醇(PVA)/酪氨酸钠(SodCa)/还原氧化石墨烯(rGO)结构的新型生物材料。通过电纺丝方法,成功地将添加了 1%、3%、5% 和 7%rGO 的 PVA/SodCa (60:40,w:w)溶液制成了抗菌有效的纳米纤维,可用作现代伤口敷料。为了创造一个使用区域,尤其是在渗出的伤口中,亲水性 PVA/SodCa/rGO 电纺垫通过浸泡在戊二醛(GLA)浴中进行交联。所有纳米纤维的表面显微照片均均匀光滑。交联后的纳米纤维完全溶于水,在蒸馏水中第 24 小时结束时,其存活率保持在 87%-81% 的范围内。据报道,这些在水环境中持续存在的生物材料表现出 275%-608% 的膨胀行为。交联后,未交联的纯 PVA/SodCa 纳米纤维的孔隙率增加了 46.75%。此外,交联的 PVA/SodCa 电纺垫的拉伸强度在 rGO 的存在下也有所增加。掺杂 3% rGO 的 PVA/SodCa 纳米纤维每 24 小时包扎一次伤口,掺杂 5% rGO 的 PVA/SodCa 纳米纤维每 48 小时包扎一次伤口,对金黄色葡萄球菌的抗菌活性分别为 99.38% 和 99.55%。
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引用次数: 0
Polymers and their engineered analogues for ocular drug delivery: Enhancing therapeutic precision 用于眼部给药的聚合物及其工程类似物:提高治疗精度
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-05 DOI: 10.1002/bip.23578
Arpon Biswas, Shivansh Kumar, Abhijit Deb Choudhury, Amol Chhatrapati Bisen, Sachin Nashik Sanap, Sristi Agrawal, Anjali Mishra, Sarvesh Kumar Verma, Mukesh Kumar, Rabi Sankar Bhatta

Ocular drug delivery is constrained by anatomical and physiological barriers, necessitating innovative solutions for effective therapy. Natural polymers like hyaluronic acid, chitosan, and gelatin, alongside synthetic counterparts such as PLGA and PEG, have gained prominence for their biocompatibility and controlled release profiles. Recent strides in polymer conjugation strategies have enabled targeted delivery through ligand integration, facilitating tissue specificity and cellular uptake. This versatility accommodates combined drug delivery, addressing diverse anterior (e.g., glaucoma, dry eye) and posterior segment (e.g., macular degeneration, diabetic retinopathy) afflictions. The review encompasses an in-depth exploration of each natural and synthetic polymer, detailing their individual advantages and disadvantages for ocular drug delivery. By transcending ocular barriers and refining therapeutic precision, these innovations promise to reshape the management of anterior and posterior segment eye diseases.

眼部给药受到解剖和生理障碍的限制,因此需要创新的解决方案来实现有效治疗。透明质酸、壳聚糖和明胶等天然聚合物以及聚乳酸(PLGA)和聚乙二醇(PEG)等合成聚合物因其生物相容性和控释特性而备受瞩目。近年来,聚合物共轭策略取得了长足进步,通过配体整合实现了靶向给药,促进了组织特异性和细胞吸收。这种多功能性可用于联合给药,解决各种前段(如青光眼、干眼症)和后段(如黄斑变性、糖尿病视网膜病变)问题。这篇综述深入探讨了每种天然和合成聚合物,详细介绍了它们在眼部给药方面的各自优缺点。通过跨越眼部障碍和提高治疗精度,这些创新有望重塑前后节眼病的治疗方法。
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引用次数: 0
Bacterial nanocellulose: A versatile biopolymer production using a cost-effective wooden disc based rotary reactor 细菌纳米纤维素:利用经济高效的木盘式旋转反应器生产多功能生物聚合物。
IF 3.2 4区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-25 DOI: 10.1002/bip.23577
Ashish Jagtap, Syed G. Dastager

Bacterial nanocellulose (BNC) has various unique qualities, including high mechanical strength, crystallinity, and high water-holding capacity, which makes it appropriate for a wide range of industrial applications. But its lower yield coupled with its high production cost creates a barrier to its usage. In this study, we have demonstrated the better yield of BNC from an indigenous strain Komagataeibacter rhaeticus MCC-0157 using a rotary disc bioreactor (RDB) having a wooden disc. The RDB was optimized based on the type of disc material, distance between the disc, and rotation speed to get the highest yield of 13.0 g/L dry material using Hestrin–Schramm (H–S) medium. Further, the bioreactor was compared for the BNC production using reported medium, which is used for static condition; the RDB showed up to fivefold increase in comparison with the static condition reported. Komagataeibacter rhaeticus MCC-0157 was previously reported to be one of the highest BNC producing stains, with 8.37 g/L of dry yield in static condition in 15 days incubation. The designed RDB demonstrated 13.0 g/L dry yield of BNC in just 5 days. Other characteristics of BNC remain same as compared with static BNC production, although the difference in the crystallinity index was observed in RDB (84.44%) in comparison with static (89.74%). For the first time, wooden disc was used for rotary bioreactor approach, which demonstrated higher yield of BNC in lesser time and can be further used for sustainable production of BNC at the industrial level.

细菌纳米纤维素(BNC)具有高机械强度、结晶性和高保水能力等各种独特品质,因此适合广泛的工业应用。但其较低的产量和较高的生产成本阻碍了它的应用。在本研究中,我们利用木质圆盘旋转盘生物反应器(RDB)从本地菌株 Komagataeibacter rhaeticus MCC-0157 中获得了较高的 BNC 产量。根据圆盘材料类型、圆盘间距离和旋转速度对 RDB 进行了优化,使用 Hestrin-Schramm (H-S) 培养基获得了 13.0 克/升的最高干物质产量。此外,该生物反应器还与用于静态条件的报告培养基的 BNC 产量进行了比较;与报告的静态条件相比,RDB 的产量增加了五倍。据报道,Komagataeibacter rhaeticus MCC-0157 是 BNC 产量最高的菌种之一,在静态条件下培养 15 天,干产量为 8.37 克/升。设计的 RDB 仅在 5 天内就显示出 13.0 克/升的 BNC 干产量。与静态 BNC 生产相比,BNC 的其他特征保持不变,但在 RDB 中观察到结晶度指数(84.44%)与静态(89.74%)不同。木盘首次用于旋转生物反应器方法,在较短时间内获得了较高的 BNC 产量,可进一步用于工业级 BNC 的可持续生产。
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引用次数: 0
Ewing Sarcoma Related protein 1 recognizes R-loops by binding DNA forks 尤文肉瘤相关蛋白 1 通过结合 DNA 叉来识别 R 环。
IF 2.9 4区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-03-21 DOI: 10.1002/bip.23576
Michelle A. Lay, Valery F. Thompson, Ajibola D. Adelakun, Jacob C. Schwartz

EWSR1 (Ewing Sarcoma Related protein 1) is an RNA binding protein that is ubiquitously expressed across cell lines and involved in multiple parts of RNA processing, such as transcription, splicing, and mRNA transport. EWSR1 has also been implicated in cellular mechanisms to control formation of R-loops, a three-stranded nucleic acid structure consisting of a DNA:RNA hybrid and a displaced single-stranded DNA strand. Unscheduled R-loops result in genomic and transcription stress. Loss of function of EWSR1 functions commonly found in Ewing Sarcoma correlates with high abundance of R-loops. In this study, we investigated the mechanism for EWSR1 to recognize an R-loop structure specifically. Using electrophoretic mobility shift assays (EMSA), we detected the high affinity binding of EWSR1 to substrates representing components found in R-loops. EWSR1 specificity could be isolated to the DNA fork region, which transitions between double- and single-stranded DNA. Our data suggests that the Zinc-finger domain (ZnF) with flanking arginine and glycine rich (RGG) domains provide high affinity binding, while the RNA recognition motif (RRM) with its RGG domains offer improved specificity. This model offers a rational for EWSR1 specificity to encompass a wide range in contexts due to the DNA forks always found with R-loops.

EWSR1(尤文肉瘤相关蛋白 1)是一种 RNA 结合蛋白,在细胞系中普遍表达,参与 RNA 处理的多个环节,如转录、剪接和 mRNA 运输。EWSR1 还与控制 R 环形成的细胞机制有关,R 环是一种三链核酸结构,由 DNA:RNA 杂交和移位的单链 DNA 链组成。计划外的 R 环会导致基因组和转录压力。尤文肉瘤中常见的 EWSR1 功能缺失与 R 环的高丰度相关。在这项研究中,我们研究了 EWSR1 特异性识别 R 环结构的机制。通过电泳迁移试验(EMSA),我们检测到 EWSR1 与代表 R 环成分的底物有高亲和力结合。EWSR1 的特异性可被分离到 DNA 叉区,该区域在双链和单链 DNA 之间转换。我们的数据表明,锌指结构域(ZnF)及其侧翼富含精氨酸和甘氨酸的结构域(RGG)提供了高亲和力的结合,而 RNA 识别基序(RRM)及其 RGG 结构域则提供了更好的特异性。这个模型为 EWSR1 的特异性提供了一个合理的解释,由于 DNA 叉总是与 R 环一起出现,因此 EWSR1 的特异性涵盖了广泛的情况。
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引用次数: 0
期刊
Biopolymers
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