Moscow University Biological Sciences Bulletin最新文献

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Enzyme and Biosurfactant Production by Exopolysaccharide Producing Bacteria Isolated from Cassava Peel Heaps 从木薯皮堆中分离出的产外多糖细菌产生的酶和生物表面活性剂

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-25 DOI: 10.3103/s0096392523600254

J. A. Amao, P. F. Omojasola, A. A. Ayandele, A. G. Adewoyin

Abstract

A major one among bacteria used in the commercial production of enzymes is Bacillus sp.; others include Escherichia sp., Pseudomonas, Proteus, Serratia, and Rhizobium, which also yield an appreciable quantity of enzymes. Biosurfactants naturally play a vital role in the teeming motility of microbes and contribute to cellular physiological procedures of signalling and, differentiation, and biofilm formation. The sourcing and selection of microbes from natural sources have endlessly been shown to be an effective way of identifying isolates of industrial value. The current study aimed to determine the ability of exopolysaccharide-producing bacteria isolated from cassava peel heap to produce some extracellular enzymes and biosurfactants. Thirteen bacteria earlier isolated from cassava peel heaps samples and found to be exopolysaccharide producers were used in this study. The highest xylanase activity was recorded at 40°C by B. amyloliquefaciens (J47). Titratable acid content was highest in peel fermented with isolate Lactobacillus plantarum (J18). In contrast, the pH of fermented peel fell below pH 7, with the lowest being the peel fermented by Pectobacterium carotovorum (J36). Three Bacillus spp. (Bacillus sp. J1, Bacillus subtilis J2, and B. amyloliquefaciens J47) were able to produce all the three enzymes (xylanase, cellulase, and β-glucosidase) assayed for. Bacterial isolates from cassava peel heap can be helpful in many industrial processes with their ability to produce substances of industrial value.

摘要在用于商业化生产酶的细菌中，主要是芽孢杆菌；其他细菌包括埃希氏菌、假单胞菌、变形杆菌、沙雷氏菌和根瘤菌，它们也产生相当数量的酶。生物表面活性剂在微生物的繁殖运动中自然发挥着重要作用，并有助于细胞的信号传递、分化和生物膜形成等生理过程。从天然资源中寻找和选择微生物，已被证明是一种有效的方法，可以鉴定出具有工业价值的分离物。本研究旨在确定从木薯皮堆中分离出来的产生外多糖的细菌产生一些胞外酶和生物表面活性剂的能力。本研究使用了早先从木薯皮堆样本中分离出的 13 种细菌，这些细菌被发现可产生外多糖。淀粉芽孢杆菌（J47）在 40°C 时的木聚糖酶活性最高。用分离的植物乳杆菌（J18）发酵的果皮中可滴定酸含量最高。相比之下，发酵果皮的 pH 值低于 pH 7，最低的是由果胶杆菌（Pectobacterium carotovorum）（J36）发酵的果皮。三个芽孢杆菌属（枯草芽孢杆菌 J1、枯草芽孢杆菌 J2 和淀粉芽孢杆菌 J47）能够产生所测定的三种酶（木聚糖酶、纤维素酶和 β-葡萄糖苷酶）。从木薯皮堆中分离出的细菌能够产生具有工业价值的物质，因此有助于许多工业过程。

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引用次数: 0

Oligoribonucleotide-Containing Nanocomplexes Based on Aminopropylsilanol Nanoparticles as Effective Inhibitors of Influenza A Virus Replication 基于氨基丙基硅烷醇纳米颗粒的含寡核苷酸纳米复合物是甲型流感病毒复制的有效抑制剂

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-25 DOI: 10.3103/s0096392524600431

M. N. Repkova, O. Yu. Mazurkov, E. I. Filippova, M. A. Procenko, N. A. Mazurkova, B. P. Chelobanov, A. S. Levina, V. F. Zarytova

Abstract

Titanium dioxide or aminopropylsilanol nanoparticles were shown to be effective vehicles for delivering oligodeoxyribonucleotides and deoxyribozymes to cells to affect target nucleic acids. In this paper, the proposed principle of the delivery has been implemented in relation to oligoribonucleotides (ORN), components of short interfering RNAs (siRNAs). It has been shown that the obtained ORN-containing nanocomplexes (Si~NH₂·ORN) based on aminopropylsilanol nanoparticles penetrate eukaryotic cells. These nanocomplexes have been investigated as agents for suppressing the replication of influenza A virus (IAV) in the cellular system. It has been shown that the ORN strands targeted to (+)RNA and (–)RNA of the IAV 5th segment reduces the titer of the virus by 99.7% and 98.4%, respectively. Thus, oligoribonucleotides in the Si~NH₂·ORN nanocomplexes effectively inhibit the replication of the influenza A virus.

摘要二氧化钛或氨丙基硅醇纳米粒子已被证明是向细胞输送寡脱氧核苷酸和脱氧核酶以影响靶核酸的有效载体。本文提出的递送原理已在短干扰 RNA（siRNA）的组成成分--寡核苷酸（ORN）中得到应用。研究表明，基于氨基丙基硅烷醇纳米粒子获得的含寡核苷酸的纳米复合物（Si~NH2-ORN）可穿透真核细胞。这些纳米复合物已被研究用于抑制甲型流感病毒（IAV）在细胞系统中的复制。研究表明，针对 IAV 第 5 节段 (+)RNA 和 (-)RNA 的 ORN 链可将病毒滴度分别降低 99.7% 和 98.4%。因此，Si~NH2-ORN 纳米复合物中的寡核苷酸能有效抑制甲型流感病毒的复制。

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引用次数: 0

Rheological Behavior of Polysaccharide Hydrogels of Alginate Reinforced by a Small Amount of Halloysite Nanotubes for Extrusion 3D Printing 海藻酸多糖水凝胶的流变行为--以少量海泡石纳米管为增强材料，用于挤压式三维打印

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700268

V. S. Molchanov, S. A. Glukhova, O. E. Philippova

Abstract

The rheological properties of hydrogels of a natural polysaccharide sodium alginate and small amount of clay nanotubes of halloysite were investigated. Changes of rheological properties during the transition from a semidiluted polymer solution to a hydrogel upon cross-linking by calcium ions were shown. In the gel state, the samples have a yield stress, and their viscosity decreases with the shear rate, but the properties are quickly recovered after the load removal. It was discovered that the addition of up to 0.3 vol % nanotubes of natural clay halloysite leads to an increase by several times of a storage modulus and a yield stress of the hydrogels. At the same time, the practically important properties of shear thinning and the rapid recovery of properties after the load removing make the nanocomposite hydrogels of alginate and halloysite nanotubes promising for use as ink for extrusion 3D printing.

摘要研究了天然多糖海藻酸钠和少量哈洛来石纳米粘土水凝胶的流变特性。结果表明，在钙离子交联作用下，半稀释聚合物溶液向水凝胶转变过程中的流变特性发生了变化。在凝胶状态下，样品具有屈服应力，其粘度随剪切速率的增加而降低，但在移除负载后，流变特性会迅速恢复。研究发现，添加多达 0.3 Vol % 的天然粘土埃洛石纳米管可使水凝胶的储存模量和屈服应力增加数倍。同时，海藻酸盐与埃洛石纳米管的纳米复合水凝胶还具有剪切稀化和卸载后快速恢复的重要特性，因此有望用作挤出三维打印的墨水。

引用次数: 0

Patterns of a Disulfide Bond Formation in Toxins of Spiders from the Ctenidae Family: A Comparison with Predictions of the AlphaFold 2.0 Neural Network 栉水母科蜘蛛毒素中二硫键的形成模式：与 AlphaFold 2.0 神经网络预测的比较

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700141

P. A. Mironov, Z. O. Shenkarev

Abstract

Venoms of wandering or running spiders from the family Ctenidae are multicomponent and include >500 identified peptides and proteins called ctenitoxins. The main components of the venom are cysteine-rich peptides containing an inhibitor cystine knot (ICK) motif. The pharmacological diversity of ctenitoxins makes it possible to consider some of them as prototypes for the development of new drugs to treat chronic pain, Huntington’s disease, erectile dysfunction, and glaucoma. Based on the localization of cysteine residues in the amino acid sequence, ctenitoxins are divided into 14 groups containing 6–14 Cys residues. To date, the spatial structure of only one ctenitoxin, ω-CNTX-Pn4a (Pha1β or Tx3-6) from the Brazilian wandering spider Phoneutria nigriventer, has been determined. Another ten structural groups of ctenitoxins are homologous to the known spatial structures of toxins from other spider families or other proteins; finally, three more ctenitoxin groups do not have any known structural homologues. In this study, we proposed possible patterns for the disulfide bond formation in all ctenitoxin groups. A comparison of the obtained schemes with the predictions of the AlphaFold 2.0 algorithm shows that this neural network does not always correctly predict the structure of cysteine-rich peptides, especially in the case of modeling of mature peptides without leader sequences.

摘要栉水母科游走性或奔跑性蜘蛛的毒液是多成分的，包括500种已确定的肽和蛋白质，称为栉水母毒素。毒液的主要成分是富含半胱氨酸的多肽，其中含有抑制胱氨酸结（ICK）基团。栉水母毒素的药理多样性使我们有可能将其中一些毒素视为开发治疗慢性疼痛、亨廷顿氏病、勃起功能障碍和青光眼新药的原型。根据半胱氨酸残基在氨基酸序列中的定位，栉水母毒素被分为 14 组，每组含有 6-14 个 Cys 残基。迄今为止，只确定了一种栉水母毒素的空间结构，即来自巴西游走蜘蛛 Phoneutria nigriventer 的 ω-CNTX-Pn4a （Pha1β 或 Tx3-6）。另外十个栉水母毒素结构组与其他蜘蛛家族或其他蛋白质的已知毒素空间结构同源；最后，还有三个栉水母毒素组没有任何已知的结构同源物。在这项研究中，我们提出了所有栉水母毒素组中二硫键形成的可能模式。将获得的方案与 AlphaFold 2.0 算法的预测结果进行比较后发现，该神经网络并不总是能正确预测富含半胱氨酸的肽的结构，尤其是在对没有前导序列的成熟肽进行建模的情况下。

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引用次数: 0

Structural Heterogeneity and Diversity of Bacillus subtilis 168 Biofilms under Different Conditions 不同条件下枯草芽孢杆菌 168 生物膜的结构异质性和多样性

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700190

T. Osire, Y. Wang, O. Burtseva, O. S. Sokolova

Abstract

Bacteria usually dwell in microbial communities of biofilms surrounded by a self-secreted extracellular matrix. This work assessed effects of different media and temperature on the development of Bacillus subtilis 168 biofilms. We applied Scanning Electron Microscopy to analyse structural differences of the pellicle biofilms grown on LB, NB, and DMS media. Significant differences were observed in biofilm morphology, and unit cell length, highlighting the significance of intricate aspects such as nutrients in bacterial/ biofilm growth. Our findings reveal the adaptation tendency of bacterial through spatial modification of its biofilm structure in response to surrounding environmental conditions such as nutrients, temperature among others, perhaps explaining variations in biofilm morphologies observed. The results present an information on biofilm diversity which could be essential for biofilm applications.

摘要细菌通常生活在由自我分泌的细胞外基质包围的生物膜微生物群落中。本研究评估了不同培养基和温度对枯草芽孢杆菌 168 生物膜发育的影响。我们使用扫描电子显微镜分析了在 LB、NB 和 DMS 培养基上生长的小柱生物膜的结构差异。我们观察到生物膜形态和单细胞长度存在显著差异，这凸显了营养物质等复杂因素在细菌/生物膜生长中的重要性。我们的研究结果揭示了细菌通过改变其生物膜结构的空间结构来适应周围环境条件（如营养物质、温度等）的趋势，这或许可以解释所观察到的生物膜形态的变化。研究结果提供了生物膜多样性的信息，这对生物膜的应用至关重要。

引用次数: 0

Cryoconveyor Protocols in Correlative Light and Electron Microscopy: From Multilevel Imaging to Modeling the Biophysical Effects and “Cryotheranostics” 相关光镜和电子显微镜中的低温输送规程：从多层次成像到生物物理效应建模和 "低温对抗 "技术

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700244

O. V. Gradov

Abstract

This paper is a technical and methodological note aimed to introduce into the practice of biological research methods a cryomicroscopy in a conveyor mode from small magnifications to the limits of magnification/resolution of scanning electron cryomicroscopy (CryoSEM). The protocol described can be applied to the samples of a low sample preparation complexity without ultratomy or processing typical of transmission electron microscopy. According to this protocol, samples are analyzed in a single microcuvette (chip) indexed by laboratory information management system and sequentially transferred from the nondestructive low-resolution optical microscopy instruments, such as lensless cryo-microscopes, to the CryoSEM/CryoESEM level (in programmable environments and atmospheres). Methods that were introduced and tested include correlative lensless cryomicroscopy and CryoSEM (including those with the sequential transition to microanalysis on wavelength-dispersive X-ray spectrometer on the Rowland circle) as well as microscopy and microinterferometry methods in the ranges from infrared to far-ultraviolet. Among the advantages of the cryoconveyor analysis protocols are sample preservation in a single portable cuvette-chip along with a possibility to establish spatial colocalization between data of optical and electron microscopy using pattern recognition software (all the way to indexing in the laboratory information system) in conducting a full range of microscopic examination. An opportunity is additionally provided for a comprehensive nondestructive sample analysis in a sequential study of microscopic systems with a possibility of variation at the subsequent stages of high-resolution microscopy, depending on the results obtained at the preceding stages of microscopy (at lower resolution).

摘要本文是一份技术和方法论说明，旨在将冷冻显微镜（CryoSEM）从小倍率到扫描电子冷冻显微镜（CryoSEM）放大倍率/分辨率极限的传输模式引入生物研究方法的实践中。所述方案适用于样品制备复杂度较低的样品，无需进行超微切片或典型的透射电子显微镜处理。根据该方案，样品在实验室信息管理系统索引的单个微量样品池（芯片）中进行分析，并按顺序从无损低分辨率光学显微镜仪器（如无镜头冷冻显微镜）转移到 CryoSEM/CryoESEM 级别（在可编程环境和气氛中）。引入和测试的方法包括相关的无透镜冷冻显微镜和 CryoSEM（包括在罗兰圈波长色散 X 射线光谱仪上依次过渡到微分析的方法），以及从红外线到远紫外线范围内的显微镜和微干涉测量方法。低温传送器分析协议的优点包括：样品保存在单个便携式比色皿芯片中，同时可以使用模式识别软件（一直到实验室信息系统中的索引）在光学和电子显微镜数据之间建立空间定位，以进行全方位的显微镜检查。此外，还提供了在显微系统的连续研究中进行全面无损样品分析的机会，根据前一阶段显微镜检查（分辨率较低）获得的结果，高分辨率显微镜检查的后续阶段有可能发生变化。

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引用次数: 0

Cryo-Electron Microscopy at the Biological Faculty of Moscow State University 莫斯科国立大学生物系的冷冻电镜技术

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700232

A. V. Moiseenko, A. M. Egorov, K. V. Shaitan, O. S. Sokolova

Abstract

The paper reports an example of a successful upgrade of the JEOL JEM-2100 analytical transmission electron microscope to a low-resolution cryo-electron microscope, which can be used to optimize sample preparation and to assess preparation quality. As a result of the instrument upgrade, it is possible to obtain subnanometer resolution of protein molecule reconstructions (within 8 Å). The role of graphene and amorphous carbon substrates in preventing the effect of preferential orientation of protein particles in a frozen sample is discussed.

摘要本文报告了将 JEOL JEM-2100 分析透射电子显微镜成功升级为低分辨率冷冻电子显微镜的实例，该仪器可用于优化样品制备和评估制备质量。仪器升级后，蛋白质分子重构可达到亚纳米分辨率（8 Å 以内）。讨论了石墨烯和无定形碳基底在防止冷冻样品中蛋白质颗粒优先取向效应方面的作用。

引用次数: 0

Structural Study of the Candida auris Ribosome 念珠菌核糖体结构研究

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700220

A. A. Atamas, A. I. Guskov, A. V. Rogachev

Abstract

Candida auris is a pathogenic fungus that causes infections in people with weakened immune systems. It is resistant to antimicrobial drugs, which complicates treatment. C. auris poses a serious threat to public health since it has high resistance and contagiousness. The ribosome plays an important role in the survival and development of this infection, and the search for inhibitors of the C. auris ribosome is of great importance. The authors used cryoelectron microscopy and single particle analysis to obtain the three-dimensional structure of the C. auris ribosome. Herein is described the architecture of the ribosome subunits and their interactions, providing valuable information for the development of novel antifungals against C. auris.

摘要念珠菌是一种致病真菌，会导致免疫力低下的人受到感染。它对抗菌药物具有抗药性，从而使治疗变得复杂。念珠菌具有很强的抗药性和传染性，对公共卫生构成严重威胁。核糖体在这种感染的生存和发展过程中起着重要作用，因此寻找 C. auris 核糖体的抑制剂非常重要。作者利用冷冻电子显微镜和单颗粒分析法获得了阿氏杆菌核糖体的三维结构。文中描述了核糖体亚基的结构及其相互作用，为开发新型抗真菌药提供了宝贵信息。

引用次数: 0

Growth of Mesenchymal Stem Cells on Oriented Microstructured Films and Electrospun Scaffolds 间充质干细胞在定向微结构薄膜和电纺支架上的生长

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700189

I. V. Demianova, E. A. Akoulina, I. I. Zharkova, V. V. Voinova, D. V. Chesnokova, A. M. Hossain, T. K. Makhina, G. A. Bonartseva, V. I. Kulikouskaya, V. V. Nikalaichuk, Yu. R. Mukhortova, A. S. Pryadko, M. A. Surmeneva, R. A. Surmenev, K. V. Shaitan, A. P. Bonartsev

Abstract

The study involved the fabrication of films with different roughness and scaffolds made of poly-3-hydroxybutyrate using various methods. Chaotic and oriented scaffolds with varying fiber thickness were obtained through the electrospinning method, depending on the polymer concentration and electrospinning parameters. Films with different surface roughness were obtained using spin coating and self-assembly methods. It was demonstrated that the varying microstructure of the surface does not affect the growth of mesenchymal stem cells over the course of 1 week; however, it does influence the morphology of the adhered cells.

摘要该研究采用各种方法制造了不同粗糙度的薄膜和聚-3-羟基丁酸酯支架。根据聚合物浓度和电纺丝参数的不同，通过电纺丝方法获得了纤维粗细不同的混沌和定向支架。利用旋涂和自组装方法获得了不同表面粗糙度的薄膜。实验证明，不同的表面微结构不会影响间充质干细胞在一周内的生长，但会影响粘附细胞的形态。

引用次数: 0

Single-Molecule Imaging of Ion Channels Isolated in Detergent and Styrene-Maleic Acid Copolymer 分离在洗涤剂和苯乙烯-马来酸共聚物中的离子通道的单分子成像

Q3 Agricultural and Biological Sciences

Moscow University Biological Sciences Bulletin

Pub Date : 2024-03-11 DOI: 10.3103/s0096392523700177

A. A. Glukhova, G. Lu, K. V. Shaitan, D. V. Bagrov, G. S. Glukhov

Abstract

In this work we purified Shaker ion channel using two alternative techniques – the detergent-based (CHAPS) and styrene-maleic acid copolymer lipid particles (XIRAN-based). Using XIRAN-based technique, we can obtain higher yield of purified protein. We used atomic force microscopy and transmission electron microscopy methods to visualize the purified single molecules. Atomic force microscopy allows measuring the diameter of the single molecules but does not reveal any high-resolution details. The sizes of individual Shaker molecules isolated by the two purification methods were close to each other.

摘要在这项工作中，我们使用了两种可供选择的技术来纯化振动台离子通道--基于去垢剂（CHAPS）和苯乙烯-马来酸共聚物脂质颗粒（基于 XIRAN）。使用基于 XIRAN 的技术，我们可以获得更高产率的纯化蛋白质。我们使用原子力显微镜和透射电子显微镜方法来观察纯化的单分子。原子力显微镜可以测量单分子的直径，但不能显示任何高分辨率的细节。两种纯化方法分离出的单个振动筛分子的大小接近。

引用次数: 0

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