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Synthesis, characterisation and in vitro anticancer activity of conjugated protease inhibitor-silver nanoparticles (AgNPs-PI) against human breast MCF-7 and prostate PC-3 cancer cell lines. 共轭蛋白酶抑制剂-银纳米粒子(AgNPs-PI)的合成、表征及对人类乳腺癌 MCF-7 和前列腺癌 PC-3 细胞株的体外抗癌活性。
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-06-01 Epub Date: 2024-05-06 DOI: 10.1007/s00449-024-03023-2
Kiran Marathe, Jitendra Naik, Vijay Maheshwari

The conjugated silver nanoparticles using biomolecules have attracted great attention of researchers because physical dimensions and surface chemistry play important roles in toxicity and biocompatibility of AgNPs. Hence, in the current study, synthesis of bio-conjugated AgNPs with protein protease inhibitor (PI) isolated from Streptomyces spp. is reported. UV-visible spectra of PI and AgNPs showed stronger peaks at 280 and 405 nm, confirming the synthesis of conjugated AgNPs-PI. TEM and SEM images of AgNPs-PI showed spherical-shaped nanoparticles with a slight increase in particle size and thin amorphous layer around the surface of silver nanomaterial. Circular dichroism, FT-IR and fluorescence spectral studies confirmed AgNPs-PI conjugation. Conjugated AgNPs-PI showed excellent anticancer potential than AgNPs and protease inhibitor separately on human breast MCF-7 and prostate PC-3 cell lines. The findings revealed that surface modification of AgNPs with protein protease inhibitor stabilised the nanomaterial and increased its anticancer activity.

由于物理尺寸和表面化学性质对银纳米粒子的毒性和生物相容性起着重要作用,因此利用生物大分子共轭银纳米粒子引起了研究人员的极大关注。因此,本研究报道了用从链霉菌中分离出的蛋白酶抑制剂(PI)合成生物共轭 AgNPs。PI 和 AgNPs 的紫外可见光谱在 280 纳米和 405 纳米处显示出较强的峰值,证实了 AgNPs-PI 的合成。AgNPs-PI 的 TEM 和 SEM 图像显示其为球形纳米颗粒,粒径略有增大,银纳米材料表面有一层薄薄的无定形层。圆二色性、傅立叶变换红外光谱和荧光光谱研究证实了 AgNPs-PI 的共轭作用。与分别使用 AgNPs 和蛋白酶抑制剂相比,共轭 AgNPs-PI 对人类乳腺 MCF-7 和前列腺 PC-3 细胞株显示出卓越的抗癌潜力。研究结果表明,用蛋白酶抑制剂对 AgNPs 进行表面修饰可稳定纳米材料并提高其抗癌活性。
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引用次数: 0
Trichoderma koningiopsis fermentation in airlift bioreactor for bioherbicide production. 在气升式生物反应器中发酵小米糠毛霉以生产生物除草剂。
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-05-01 Epub Date: 2024-03-30 DOI: 10.1007/s00449-024-02991-9
Aline Frumi Camargo, Simone Kubeneck, Charline Bonatto, Suzana Fátima Bazoti, Júlia Pieper Nerling, Gabriel Henrique Klein, William Michelon, Sérgio L Alves, Altemir José Mossi, Gislaine Fongaro, Helen Treichel

During scaling of fermentations, choosing a bioreactor is fundamental to ensure the product's quality. This study aims to produce bioherbicides using Trichoderma koningiopsis fermentation, evaluating process parameters in an Airlift bioreactor. As a response, we quantified the production of enzymes involved in the bioherbicide activity (amylase, cellulase, laccase, lipase, and peroxidase). In addition, it evaluated the agronomic efficiency of the fermented extract optimized through tests that promoted soybean growth and nodulation, soybean seed germination, and in vitro phytopathogen control. As a result of optimizing the scaling bioprocess, it was possible to obtain an adequate fermentation condition, which, when applied to soybean seeds, had beneficial effects on their growth. It allowed the production of an enzyme cocktail. These results add a crucial biotechnological potential factor for the success of the optimized formulation in the Airlift bioreactor, in addition to presenting relevant results for the scientific community.

在扩大发酵规模的过程中,选择生物反应器是确保产品质量的关键。本研究旨在利用科宁氏毛霉发酵法生产生物除草剂,评估气升式生物反应器的工艺参数。作为回应,我们对参与生物除草剂活性的酶(淀粉酶、纤维素酶、漆酶、脂肪酶和过氧化物酶)的产量进行了量化。此外,我们还通过促进大豆生长和结瘤、大豆种子发芽以及体外植物病原体控制等试验,评估了发酵提取物的农艺效率。通过优化缩放生物工艺,获得了适当的发酵条件。它允许生产一种鸡尾酒酶。除了向科学界展示相关成果外,这些成果还为空运生物反应器中优化配方的成功增加了一个重要的生物技术潜在因素。
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引用次数: 0
Improved production of RNA-inhibiting antimicrobial peptide by Bacillus licheniformis MCC 2514 facilitated by a genetic algorithm optimized medium. 遗传算法优化培养基促进地衣芽孢杆菌 MCC 2514 生产 RNA 抑制抗菌肽。
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-05-01 Epub Date: 2024-03-23 DOI: 10.1007/s00449-024-02998-2
Ishrat Jahan Peerzade, Sarma Mutturi, Prakash M Halami

One of the significant challenges during the purification and characterization of antimicrobial peptides (AMPs) from Bacillus sp. is the interference of unutilized peptides from complex medium components during analytical procedures. In this study, a semi-synthetic medium was devised to overcome this challenge. Using a genetic algorithm, the production medium of AMP is optimized. The parent organism, Bacillus licheniformis MCC2514, produces AMP in very small quantities. This AMP is known to inhibit RNA biosynthesis. The findings revealed that lactose, NH4Cl and NaNO3 were crucial medium constituents for enhanced AMP synthesis. The potency of the AMP produced was studied using bacterium, Kocuria rhizophila ATCC 9341. The AMP produced from the optimized medium was eightfold higher than that produced from the unoptimized medium. Furthermore, activity was increased by 1.5-fold when cultivation conditions were standardized using the optimized medium. Later, AMP was produced in a 5 L bioreactor under controlled conditions, which led to similar results as those of shake-flask production. The mode of action of optimally produced AMP was confirmed to be inhibition of RNA biosynthesis. Here, we demonstrate that improved production of AMP is possible with the developed semi-synthetic medium recipe and could help further AMP production in an industrial setup.

在纯化和表征芽孢杆菌抗菌肽(AMPs)的过程中,面临的重大挑战之一是分析过程中复杂培养基成分对未利用肽的干扰。本研究设计了一种半合成培养基来克服这一难题。利用遗传算法优化了 AMP 的生产培养基。母体地衣芽孢杆菌 MCC2514 能产生极少量的 AMP。众所周知,这种 AMP 能抑制 RNA 的生物合成。研究结果表明,乳糖、NH4Cl 和 NaNO3 是增强 AMP 合成的关键培养基成分。使用 Kocuria rhizophila ATCC 9341 细菌研究了所产生的 AMP 的效力。优化培养基产生的 AMP 是未优化培养基的八倍。此外,当使用优化培养基对培养条件进行标准化后,其活性提高了 1.5 倍。随后,在受控条件下,在 5 升生物反应器中生产 AMP,结果与摇瓶生产类似。经证实,优化生产的 AMP 的作用模式是抑制 RNA 的生物合成。在此,我们证明了使用所开发的半合成培养基配方可以提高 AMP 的产量,并有助于在工业装置中进一步生产 AMP。
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引用次数: 0
The function of microbial enzymes in breaking down soil contaminated with pesticides: a review. 微生物酶在分解受农药污染的土壤中的功能:综述。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-05-01 Epub Date: 2024-03-08 DOI: 10.1007/s00449-024-02978-6
Xing Kai Chia, Tony Hadibarata, Risky Ayu Kristanti, Muhammad Noor Hazwan Jusoh, Inn Shi Tan, Henry Chee Yew Foo

The use of pesticides and the subsequent accumulation of residues in the soil has become a worldwide problem. Organochlorine (OC) pesticides have spread widely in the environment and caused contamination from past agricultural activities. This article reviews the bioremediation of pesticide compounds in soil using microbial enzymes, including the enzymatic degradation pathway and the recent development of enzyme-mediated bioremediation. Enzyme-mediated bioremediation is divided into phase I and phase II, where the former increases the solubility of pesticide compounds through oxidation-reduction and hydrolysis reactions, while the latter transforms toxic pollutants into less toxic or nontoxic products through conjugation reactions. The identified enzymes that can degrade OC insecticides include dehalogenases, phenol hydroxylase, and laccases. Recent developments to improve enzyme-mediated bioremediation include immobilization, encapsulation, and protein engineering, which ensure its stability, recyclability, handling and storage, and better control of the reaction.

农药的使用及其在土壤中的残留积累已成为一个世界性问题。有机氯(OC)杀虫剂在环境中广泛传播,并在过去的农业活动中造成污染。本文综述了利用微生物酶对土壤中的农药化合物进行生物修复的情况,包括酶降解途径和酶介导生物修复的最新发展。酶介导生物修复分为第一阶段和第二阶段,前者通过氧化还原和水解反应增加农药化合物的溶解度,后者通过共轭反应将有毒污染物转化为毒性较低或无毒的产物。已发现的可降解 OC 杀虫剂的酶包括脱卤酶、苯酚羟化酶和裂解酶。最近在改进酶介导的生物修复方面取得的进展包括固定化、封装和蛋白质工程,这确保了酶的稳定性、可回收性、处理和储存,以及对反应的更好控制。
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引用次数: 0
Analyzing of hydrodynamic stress and mass transfer requirements of a fermentation process carried out in a coaxial bioreactor: a scale-up study. 分析在同轴生物反应器中进行发酵过程的流体动力应力和传质要求:规模扩大研究。
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-05-01 Epub Date: 2024-04-01 DOI: 10.1007/s00449-024-02990-w
Ali Rahimzadeh, Farhad Ein-Mozaffari, Ali Lohi

Fluid hydrodynamic stress has a deterministic effect on the morphology of filamentous fungi. Although the coaxial mixer has been recognized as a suitable gas dispersion system for minimizing inhomogeneities within a bioreactor, its performance for achieving enhanced oxygen transfer while operating at a reduced shear environment has not been investigated yet, specifically upon scale-up. Therefore, the influence of the impeller type, aeration rate, and central impeller retrofitting on the efficacy of an abiotic coaxial system containing a shear-thinning fluid was examined. The aim was to assess the hydrodynamic parameters, including stress, mass transfer, bubble size, and gas hold-up, upon conducting a scale-up study. The investigation was conducted through dynamic gassing-in, tomography, and computational fluid dynamics combined with population balance methods. It was observed that the coaxial bioreactor performance was strongly influenced by the agitator type. In addition, coaxial bioreactors are scalable in terms of shear environment and oxygen transfer rate.

流体流体动力对丝状真菌的形态具有决定性影响。虽然同轴混合器已被认为是一种合适的气体分散系统,可最大限度地减少生物反应器内的不均匀性,但其在减少剪切力的环境中运行时增强氧气转移的性能尚未得到研究,特别是在放大时。因此,我们研究了叶轮类型、曝气速率和中央叶轮改装对含有剪切稀化流体的非生物同轴系统功效的影响。目的是在进行放大研究时评估流体力学参数,包括应力、传质、气泡大小和气体截留。研究通过动态进气、层析成像和计算流体力学结合种群平衡方法进行。研究发现,同轴生物反应器的性能受搅拌器类型的影响很大。此外,同轴生物反应器在剪切环境和氧气转移率方面具有可扩展性。
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引用次数: 0
Correction: The function of microbial enzymes in breaking down soil contaminated with pesticides: a review. 更正:微生物酶在分解受农药污染的土壤中的功能:综述。
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-05-01 DOI: 10.1007/s00449-024-03026-z
Xing Kai Chia, Tony Hadibarata, Risky Ayu Kristanti, Muhammad Noor Hazwan Jusoh, Inn Shi Tan, Henry Chee Yew Foo
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引用次数: 0
Kinetic modeling and optimization of the mono- and diglycerides synthesis mediated by the lipase Lipozyme® TL 100 L immobilized on clayey support. 固定在粘土支撑物上的脂肪酶 Lipozyme® TL 100 L 介导的单甘油酯和二甘油酯合成的动力学建模和优化。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-05-01 Epub Date: 2024-03-27 DOI: 10.1007/s00449-024-02999-1
George F Finco, Edson A da Silva, Fernando Palú, Márcia R F Klen, Karina G Fiametti, João H C Wancura, J Vladimir Oliveira

Mono- and diglycerides play a crucial role in the food industry as multifunctional food additives and emulsifiers. Their importance stems from their unique properties, which allow them to improve the quality, texture, and stability of various food products. Here, results of the kinetic modeling of the mono- and diglycerides synthesis mediated by the lipase Lipozyme® TL 100 L immobilized on the clayey support Spectrogel® type C are reported. The support was characterized by TEM, SEM, and FTIR. Firstly, the influence of pH and lipase load on the immobilization process was analyzed, resulting in an enzymatic activity of 93.2 ± 0.7 U g-1 under optimized conditions (170.9 U g-1 of lipase and pH of 7.1). Afterward, the effects of reaction temperature and concentration of immobilized biocatalyst in the feedstock conversion were evaluated. At optimized parameters, a triglycerides conversion of 97% was obtained at 36.5 °C, 7.9 vol.% of enzyme, a glycerol to feedstock molar ratio of 2:1, and 2 h. The optimized conditions were used to determine the kinetic constants of the elementary reactions involved in the glycerolysis, where a fit superior to 0.99 was achieved between experimental values and predicted data.

单甘酯和二甘酯作为多功能食品添加剂和乳化剂,在食品工业中发挥着至关重要的作用。它们的重要性源于其独特的特性,这些特性使它们能够改善各种食品的质量、质地和稳定性。本文报告了固定在粘土支撑 Spectrogel® C 型上的脂肪酶 Lipozyme® TL 100 L 介导的单甘油酯和二甘油酯合成动力学模型的结果。透射电镜、扫描电镜和傅立叶变换红外光谱对该载体进行了表征。首先,分析了 pH 值和脂肪酶载量对固定化过程的影响,结果表明在优化条件下(脂肪酶 170.9 U g-1,pH 值为 7.1),酶活性为 93.2 ± 0.7 U g-1。随后,评估了反应温度和固定化生物催化剂浓度对原料转化的影响。根据优化参数,在温度为 36.5 °C、酶浓度为 7.9 vol.%、甘油与原料摩尔比为 2:1、反应时间为 2 小时的条件下,甘油三酯转化率达到 97%。
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引用次数: 0
Dual stress factors adaptive evolution for high EPA production in Schizochytrium sp. and metabolomics mechanism analysis Schizochytrium sp.高产 EPA 的双重压力因素适应性进化及代谢组学机制分析
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-04-30 DOI: 10.1007/s00449-024-03013-4
Ying Ou, Yu Qin, Shoushuai Feng, Hailin Yang

Eicosapentaenoic acid (EPA) is a vital ɷ-3 polyunsaturated fatty acid (PUFA) for human body with various physiological functions. In this study, we proposed an adaptive evolutionary strategy based on high-temperature and high-oxygen two-factor stress to increase the EPA production capacity of Schizochytrium. High-temperature stress was used to increase EPA yield, and high oxygen was implemented to continuously stimulate cell growth and lipid accumulation. The biomass and EPA production of ALE-D50 reached 35.33 g/L and 1.54 g/L, which were 43.85% and 71.11% higher than that of the original strain, respectively. Lower in vivo reactive oxygen species levels indicated that the evolved strain possessed stronger antioxidant activity. Liquid chromatography–mass spectrometry metabolomics showed that enhanced glucose consumption and glycolysis metabolism, as well as a weakened tricarboxylic acid cycle and reduced amino acid metabolic tributaries in the evolved strain, might be associated with increased growth and EPA synthesis. Finally, the lipid production and EPA production in a fed-batch fermentation were further increased to 48.93 g/L and 3.55 g/L, improving by 54.30% and 90.86%, respectively. This study provides a novel pathway for promoting EPA biosynthesis in Schizochytrium.

二十碳五烯酸(EPA)是人体重要的ɷ-3多不饱和脂肪酸(PUFA),具有多种生理功能。本研究提出了一种基于高温和高氧双因子胁迫的适应性进化策略,以提高裂头藻的 EPA 产能。高温胁迫用于提高 EPA 产量,高氧则用于持续刺激细胞生长和脂质积累。ALE-D50的生物量和EPA产量分别达到35.33克/升和1.54克/升,比原始菌株分别高出43.85%和71.11%。体内活性氧水平的降低表明进化菌株具有更强的抗氧化活性。液相色谱-质谱代谢组学研究表明,进化株的葡萄糖消耗和糖酵解代谢增强,三羧酸循环减弱,氨基酸代谢支流减少,这可能与进化株的生长和 EPA 合成增加有关。最后,在饲料批量发酵中,脂质产量和 EPA 产量进一步提高到 48.93 克/升和 3.55 克/升,分别提高了 54.30% 和 90.86%。这项研究为促进裂头蚴中 EPA 的生物合成提供了一条新途径。
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引用次数: 0
Promoting mechanism of nitrogen removal by Fe3O4 magnetic particles during the start-up phase in sequencing batch reactor 序批式反应器启动阶段 Fe3O4 磁性颗粒的脱氮促进机制
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-04-27 DOI: 10.1007/s00449-024-03006-3
Xiaoyu Hu, Shuai Zhang, Guicai Liu, Jiabin Wang, Yue Wang

In this paper, a magnetic sequencing batch reactor (SBR) was constructed, and the influence rule of magnetic particle dosing performance of denitrification was investigated. The diversity, structure, and potential functions of the microbial community were comprehensively explored. The results showed that the particle size and the dosage of Fe3O4 magnetic particles were the main parameters affecting the sedimentation performance of activated sludge. The start-up phase of the SBR reactor with Fe3O4 magnetic particles was 5 days less than the control. Moreover, total nitrogen removal efficiency during the start-up phase was improved, with the maximum value reaching 91.93%, surpassing the control by 9.7% with the Fe3O4 dosage of 1.2 g L−1. In addition, the activated sludge concentration and dehydrogenase activity were improved, compared to the control. High-throughput sequencing showed that the denitrifying bacterium Saccharimonadales dominated the reactor and was enriched by magnetic particles. According to predicted functions, the abundance of genes for denitrification increased with the addition of magnetic particles, suggesting the capacity of nitrogen removal was enhanced in the microbial community. Overall, the Fe3O4 magnetic particles provide great potential for enhanced wastewater nitrogen removal.

Graphical Abstract

本文构建了磁性序批式反应器(SBR),并研究了磁性颗粒加药对反硝化性能的影响规律。全面探讨了微生物群落的多样性、结构和潜在功能。结果表明,粒径和 Fe3O4 磁性颗粒的投加量是影响活性污泥沉降性能的主要参数。使用 Fe3O4 磁性颗粒的 SBR 反应器的启动阶段比对照组少 5 天。此外,在 Fe3O4 投加量为 1.2 g L-1 的情况下,启动阶段的总氮去除率有所提高,最大值达到 91.93%,比对照组高出 9.7%。此外,与对照组相比,活性污泥浓度和脱氢酶活性也有所提高。高通量测序显示,反硝化细菌 Saccharimonadales 在反应器中占主导地位,并通过磁性颗粒得到富集。根据预测功能,反硝化基因的丰度随着磁性颗粒的添加而增加,这表明微生物群落的脱氮能力得到了提高。总之,Fe3O4 磁性颗粒在提高废水脱氮能力方面具有巨大潜力。 图文摘要
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引用次数: 0
Identification of hyperthermophilic D-allulose 3-epimerase from Thermotoga sp. and its application as a high-performance biocatalyst for D-allulose synthesis 嗜热菌 D-allulose 3-epimerase 的鉴定及其在 D-allulose 合成中作为高性能生物催化剂的应用
IF 3.8 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-04-27 DOI: 10.1007/s00449-024-02989-3
Ji-Dong Shen, Bao-Ping Xu, Te-Li Yu, Yong-Xiang Fei, Xue Cai, Liang-Gang Huang, Li-Qun Jin, Zhi-Qiang Liu, Yu-Guo Zheng

D-Allulose 3-epimerase (DAE) is a vital biocatalyst for the industrial synthesis of D-allulose, an ultra-low calorie rare sugar. However, limited thermostability of DAEs hinders their use at high-temperature production. In this research, hyperthermophilic TI-DAE (Tm = 98.4 ± 0.7 ℃) from Thermotoga sp. was identified via in silico screening. A comparative study of the structure and function of site-directed saturation mutagenesis mutants pinpointed the residue I100 as pivotal in maintaining the high-temperature activity and thermostability of TI-DAE. Employing TI-DAE as a biocatalyst, D-allulose was produced from D-fructose with a conversion rate of 32.5%. Moreover, TI-DAE demonstrated excellent catalytic synergy with glucose isomerase CAGI, enabling the one-step conversion of D-glucose to D-allulose with a conversion rate of 21.6%. This study offers a promising resource for the enzyme engineering of DAEs and a high-performance biocatalyst for industrial D-allulose production.

D- Allulose 3-epimerase(DAE)是工业合成超低卡路里稀有糖类 D- Allulose 的重要生物催化剂。然而,DAE 的热稳定性有限,阻碍了其在高温生产中的应用。在这项研究中,通过硅学筛选,从Thermotoga sp.中鉴定出了超嗜热TI-DAE(Tm = 98.4 ± 0.7 ℃)。通过对定点饱和突变突变体的结构和功能进行比较研究,发现残基 I100 是维持 TI-DAE 高温活性和耐热性的关键。使用 TI-DAE 作为生物催化剂,可以从 D-果糖生产出 D-阿洛酮糖,转化率高达 32.5%。此外,TI-DAE 与葡萄糖异构酶 CAGI 表现出了极佳的催化协同作用,可将 D-葡萄糖一步转化为 D-阿洛糖,转化率为 21.6%。这项研究为 DAE 的酶工程提供了一种前景广阔的资源,也为工业化生产 D-阿洛糖提供了一种高性能生物催化剂。
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引用次数: 0
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Bioprocess and Biosystems Engineering
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