Biochemical Engineering Journal最新文献_第4页

Development of inducible packaging cell line for rAAV production via CRISPR-Cas9 mediated site-specific integration 通过 CRISPR-Cas9 介导的位点特异性整合，开发用于生产 rAAV 的可诱导包装细胞系

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-31 DOI: 10.1016/j.bej.2024.109552

Qiang Fu , Yongdan Wang , Emily Doleh , Mark Blenner , Seongkyu Yoon

AAV-mediated gene therapy is a quickly growing segment of the pharmaceutical market; however, the current transient transfection process to produce rAAV has several challenges. The stable cells are ideal for large-scale continuous production, overcoming the drawbacks in the current transient transfection and streamlining rAAV production. In this study, we proposed to use synthetic inducible promoters to control the viral component expression and develop the baseline of HEK293T stable cells via site-specific integration mediated with CRISPR-Cas9, targeting safe harbor sites of human genome (ROSA26, AAVS1, and CCR5 locus). With a total of three round integrations, stable cell pools were developed and evaluated at each round of integration. Single clones were further characterized for each integration round. Regarding the stable pools, the 5’ and 3’ junction PCR results confirmed the site-specific integration to each locus. The genome copy result showed that AAV components, including Rep78/68, E2A, E4orf6, Cap, and Rep52/40, were successfully integrated into the host cell genome. Genome and capsid titer after induction confirmed rAAV production for stable cell pools in each round. The packaging cell line (after 2nd round integration) was able to produce rAAV. However, it was observed that the genome titer was ten-fold lower than that of rAAV products done with triple plasmids transfection. The out-to-out PCR and qPCR assay results further confirm the site-specific integration. This research demonstrates the feasibility of developing the inducible stable cell line with the refactored viral vectors via a site-specific integration.

AAV 介导的基因疗法是医药市场中一个快速增长的细分市场；然而，目前生产 rAAV 的瞬时转染工艺面临着一些挑战。稳定细胞是大规模连续生产的理想选择，它克服了目前瞬时转染的缺点，简化了 rAAV 的生产过程。在本研究中，我们提出使用合成诱导启动子来控制病毒成分的表达，并通过 CRISPR-Cas9 介导的位点特异性整合，以人类基因组的安全港位点（ROSA26、AAVS1 和 CCR5 位点）为目标，开发 HEK293T 稳定细胞基线。总共进行了三轮整合，在每一轮整合中都开发并评估了稳定细胞池。对每轮整合的单克隆进行了进一步鉴定。关于稳定细胞池，5'和 3'连接PCR结果证实了每个基因座的特异性整合。基因组拷贝结果显示，包括 Rep78/68、E2A、E4orf6、Cap 和 Rep52/40 在内的 AAV 成分已成功整合到宿主细胞基因组中。诱导后的基因组和囊壳滴度证实，每一轮都有稳定的细胞池产生了 rAAV。包装细胞系（第二轮整合后）能够产生 rAAV。但观察到基因组滴度比三重质粒转染的 rAAV 产物低十倍。从外到内的 PCR 和 qPCR 检测结果进一步证实了位点特异性整合。这项研究证明了通过位点特异性整合利用重构病毒载体开发可诱导的稳定细胞系的可行性。

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引用次数: 0

Impact of tetracycline on mixotrophic denitrification process under different sulfur to nitrogen ratios 不同硫氮比条件下四环素对混养反硝化过程的影响

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-31 DOI: 10.1016/j.bej.2024.109557

Bohan Lv , Yang-Guo Zhao , Yue Chen , Mupindu Progress , Mengchun Gao , Liang Guo , Junyuan Ji , Chunji Jin

The sulfur-based autotrophic-heterotrophic denitrification, i.e., mixotrophic denitrification, is suitable for the nitrate and antibiotics removal in aquaculture tailwater at a low COD to nitrogen (C/N) ratio. This study focused on the effect of tetracycline (TC) on mixotrophic denitrification under different S/N ratios. Two bioreactors were simultaneously operated with or without dosing tetracycline under different sulfur to nitrogen (S/N) ratios of 3.94, 4.64 and 5.94. The results showed that the removal rate of total inorganic nitrogen (TIN) increased from 0.25 to 0.69 mg N L⁻¹ min⁻¹ with the rise of S/N ratio, while TC dosage significantly declined the removal efficiency of TIN. Dissimilatory nitrogen reduction to ammonia (DNRA) bacteria was detected when exposing to TC, indicating that DNRA presented more resistance to TC. The removal efficiency of TC in the denitrification system reached the maximum of 22.87 % at S/N of 4.64. Meanwhile the genus Marinicella was detected at this phase, which was conducive to the degradation of organic pollutants. This study found that TC promoted the accumulation of ammonia nitrogen, and had a great effect on sulfur autotrophic bacteria at S/N of 5.94. The removal of TC mainly depended on microbial co-metabolism, and there was a significant correlation between the reduction of TC concentration and the decrease of sulfur compounds (p < 0.05). 4.64 is the best S/N ratio for the mixotrophic denitrification process, which revealed maximum nitrate and TC removal rates.

以硫为基础的自养-异养反硝化作用，即混养反硝化作用，适用于在低化学需氧量（COD）-氮（C/N）比条件下去除养殖尾水中的硝酸盐和抗生素。本研究主要探讨了四环素（TC）在不同信噪比条件下对混养反硝化作用的影响。在不同的硫氮（S/N）比（3.94、4.64 和 5.94）条件下，同时运行两个生物反应器，是否投加四环素。结果表明，随着硫氮比的升高，总无机氮（TIN）的去除率从 0.25 mg N L-1 min-1 提高到 0.69 mg N L-1 min-1，而四环素的投加量则显著降低了总无机氮的去除效率。在暴露于 TC 的情况下，检测到了分解氮还原氨（DNRA）细菌，这表明 DNRA 对 TC 有更强的抵抗力。反硝化系统对 TC 的去除率在信噪比为 4.64 时达到最高，为 22.87%。同时，在这一阶段检测到了马林杆菌属，这有利于有机污染物的降解。本研究发现，在信噪比为 5.94 时，TC 促进了氨氮的积累，并对硫自养菌有很大影响。TC 的去除主要依赖于微生物的协同代谢，TC 浓度的降低与硫化合物的减少之间存在显著相关性（p <0.05）。4.64 是混养反硝化过程的最佳信噪比，它显示了最大的硝酸盐和 TC 去除率。

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引用次数: 0

Pros and cons of airlift and bubble column bioreactors: How internals improve performance 气浮和气泡柱生物反应器的优缺点：内部结构如何提高性能

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-30 DOI: 10.1016/j.bej.2024.109539

Carolin Bokelmann , Jason Bromley , Ralf Takors

Gas fermentation is a promising technology of high commercial interest, particularly for capturing CO₂ and CO from industrial off-gases to reduce greenhouse gas emissions and replace fossil fuels for bulk chemical production. Therefore, evaluating promising bioreactor settings ab initio is a crucial step. Whereas alternate configurations may be tested in laborious scale up studies, the procedure may be accelerated by in silico studies that accompany or even partially replace wet-lab work once the models are validated. In this context, the current study compares various pneumatically agitated reactor types – bubble column reactor (BCR), annulus- and center-rising internal-loop airlift reactor (AR-IL-ALR and CR-IL-ALR), and external-loop airlift reactor (EL-ALR) – to identify advantages and disadvantages for the given application based on computational fluid dynamics (CFD) models. Process performance is optimized by introducing internal structures to guide the flow. Despite a significant increase in the mass transfer coefficient (

k_{L} a

) through internal modifications, the CR-IL-ALR still exhibited the poorest performance. The optimized AR-IL-ALR demonstrated good mixing and, after introducing an open-cone shaped internal in the head part and a conical bottom, superior mass transfer, achieving an enhancement over 10 % in the mass transfer coefficient to 315 1/h. This study thereby outlines the potential of internal structures for process improvement, as well as the value of a priori in silico design of reactor configurations.

气体发酵是一项极具商业价值的技术，特别是从工业废气中捕获二氧化碳和一氧化碳，以减少温室气体排放，并取代化石燃料用于大宗化学品生产。因此，对有前景的生物反应器设置进行初始评估是至关重要的一步。虽然可以在费力的放大研究中测试替代配置，但在模型得到验证后，可以通过伴随甚至部分取代湿实验室工作的硅学研究来加速这一过程。在这种情况下，当前的研究比较了各种气动搅拌反应器类型--气泡塔反应器（BCR）、环形和中心上升内循环气动反应器（AR-IL-ALR 和 CR-IL-ALR）以及外循环气动反应器（EL-ALR）--根据计算流体动力学（CFD）模型确定特定应用的优缺点。通过引入内部结构来引导气流，从而优化工艺性能。尽管通过内部改造大大提高了传质系数（kLa），但 CR-IL-ALR 的性能仍然最差。优化后的 AR-IL-ALR 不仅混合效果良好，而且在头部引入开放式锥形内部结构和锥形底部后，传质效果更佳，传质系数提高了 10% 以上，达到 315 1/h。这项研究由此概述了内部结构在改进工艺方面的潜力，以及对反应器配置进行先验硅设计的价值。

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引用次数: 0

CRISPR/Cas9-mediated knockout of DYRK1B in triple-negative breast cancer cells: implications for cell proliferation, apoptosis, and therapeutic sensitivity CRISPR/Cas9 介导的三阴性乳腺癌细胞 DYRK1B 基因敲除：对细胞增殖、凋亡和治疗敏感性的影响

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-29 DOI: 10.1016/j.bej.2024.109553

Asrin Rashidi , Ernst-Martin Füchtbauer , Zakaria Vahabzadeh , Farzad Soleimani , Karim Rahimi , Bahram Nikkhoo , Shohreh Fakhari , Mohammad Bagher Khadem Erfan , Asaad Azarnezhad , Arash Pooladi , Fariborz Soheili , Fardin Fathi

Breast cancer is the most common cancer among women worldwide, with the triple-negative subtype (TNBC) having a poor prognosis and limited treatment options. DYRK1B is a dual-specificity kinase that regulates the cell cycle and quiescence. While its role in several cancers has been characterized, its role in TNBC remains unknown. In this study, we used CRISPR/Cas9 to delete DYRK1B in MDA-MB-231 cells, a model of TNBC and investigated its effects on cell proliferation, apoptosis, invasion, migration, angiogenesis, and response to Paclitaxel. The DYRK1B knockout (KO) was confirmed by PCR, Real-time qPCR, and Sanger sequencing. KO cells showed a significant reduction in cell proliferation, colony formation, invasion, and migration. Additionally, there were alterations in mRNA expression levels of several genes related to the cell cycle, angiogenesis, and cell motility, such as CCND1, MCM2, PCNA, CDKN1B, HIF1A, VEGFA, and WASF3, compared to MDA-MB-231 wild type (WT) cells. Immunocytochemistry results assessing Ki67 expression, a marker of cell proliferation, indicated that DYRK1B knockout cells had significantly lower Ki67 expression than WT cells. Furthermore, KO cells exhibited increased apoptosis and sensitivity to contact inhibition. Additionally, the IC₅₀ for Paclitaxel was significantly decreased in KO cells. These results suggest that DYRK1B plays an important role in the survival and invasion of TNBC cells and might be a potential candidate as a new therapeutic target for this disease.

乳腺癌是全球妇女最常见的癌症，其中三阴性亚型（TNBC）预后较差，治疗方案有限。DYRK1B 是一种双重特异性激酶，可调节细胞周期和静止状态。虽然DYRK1B在多种癌症中的作用已被证实，但它在TNBC中的作用仍不清楚。在本研究中，我们使用 CRISPR/Cas9 技术在 TNBC 模型 MDA-MB-231 细胞中删除了 DYRK1B，并研究了它对细胞增殖、凋亡、侵袭、迁移、血管生成和对紫杉醇反应的影响。通过 PCR、实时 qPCR 和 Sanger 测序证实了 DYRK1B 基因敲除（KO）。KO细胞在细胞增殖、集落形成、侵袭和迁移方面均表现出明显的下降。此外，与 MDA-MB-231 野生型（WT）细胞相比，与细胞周期、血管生成和细胞运动相关的几个基因（如 CCND1、MCM2、PCNA、CDKN1B、HIF1A、VEGFA 和 WASF3）的 mRNA 表达水平也发生了变化。免疫细胞化学评估细胞增殖标志物 Ki67 表达的结果表明，DYRK1B 基因敲除细胞的 Ki67 表达明显低于 WT 细胞。此外，KO 细胞表现出更强的凋亡能力和对接触抑制的敏感性。此外，KO 细胞中紫杉醇的 IC50 值明显降低。这些结果表明，DYRK1B在TNBC细胞的存活和侵袭过程中起着重要作用，可能成为治疗这种疾病的新靶点。

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引用次数: 0

Cell Dome-based transfection array for non-adherent suspension cells 基于细胞穹顶的非粘附悬浮细胞转染阵列

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-29 DOI: 10.1016/j.bej.2024.109554

Ryotaro Kazama , Satoshi Fujita , Shinji Sakai

Cell-based microarrays are valuable tools for analyzing cellular functions. However, a significant limitation of conventional microarrays is their inapplicability to non-adherent cells. In this study, we investigated the potential of the ‘Cell Dome’ (diameter: 1 mm, height: approximately 300 µm) with a 90 µm-thick hydrogel shell as a gene transfection array for non-adherent cells in suspension. The human lymphoma cell line (K562 cells), used as a model for non-adherent cells, was transfected more efficiently by Lipofectamine/pDNA complexes on a composite hydrogel made of polyvinyl alcohol derivative (PVA-Ph) and chitosan derivative (chitosan-Ph) than hydrogels composed of an alginate derivative or PVA-Ph alone. Moreover, no significant adverse effects on the viability and proliferation of the enclosed cells were observed for Cell Dome with a PVA-Ph/chitosan-Ph composite hydrogel shell. Lipofectamine/pDNA complexes released from the bottom of Cell Domes could transfect the enclosed cells without leaking or contaminating adjacent Cell Domes. These results demonstrate the potential of Cell Domes with an appropriate hydrogel shell as transfection arrays for non-adherent cells in suspension, thereby expanding the range of applications of cell-based array technologies. This novel Cell-Dome transfection array would be a valuable tool for analyzing the cellular function of non-adherent cells in suspension and showcases the potential for providing important biomedical insights for future research and developments.

基于细胞的芯片是分析细胞功能的重要工具。然而，传统微阵列的一个重要局限是不适用于非粘附细胞。在这项研究中，我们研究了带有 90 微米厚水凝胶外壳的 "细胞穹顶"（直径：1 毫米，高度：约 300 微米）作为基因转染阵列用于悬浮非粘附细胞的潜力。以人类淋巴瘤细胞株（K562 细胞）为非粘附细胞模型，在由聚乙烯醇衍生物（PVA-Ph）和壳聚糖衍生物（壳聚糖-Ph）组成的复合水凝胶上，脂质体转染胺/pDNA 复合物的转染效率要高于单独由海藻酸衍生物或 PVA-Ph 组成的水凝胶。此外，带有 PVA-Ph/ 壳聚糖-Ph 复合水凝胶外壳的 Cell Dome 对被包裹细胞的活力和增殖没有明显的不良影响。从细胞穹顶底部释放的脂质体/pDNA 复合物可以转染被封闭的细胞，而不会泄漏或污染相邻的细胞穹顶。这些结果表明，带有适当水凝胶外壳的细胞穹顶具有作为悬浮非粘附细胞转染阵列的潜力，从而扩大了基于细胞的阵列技术的应用范围。这种新型细胞穹顶转染阵列将成为分析悬浮液中非粘附细胞功能的重要工具，并有可能为未来的研究和开发提供重要的生物医学见解。

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引用次数: 0

Removal of lysozyme as protein waste using weak ion exchange nanofiber membrane in a batch system: Linear and nonlinear model analysis 在批处理系统中使用弱离子交换纳米纤维膜去除作为蛋白质废物的溶菌酶：线性和非线性模型分析

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-29 DOI: 10.1016/j.bej.2024.109550

Dinh Thi Hong Thanh , Nguyen The Duc Hanh , Bing-Lan Liu , Penjit Srinophakun , Chen-Yaw Chiu , Shen-Long Tsai , Kuei-Hsiang Chen , Yu-Kaung Chang

This study investigates the removal of lysozyme, a common waste protein, using weak ion-exchange nanofiber membranes (P-COOH) to address challenges in protein waste management and wastewater treatment. Protein waste, prevalent in industrial and biological processes, contributes to environmental pollution and increases treatment complexity if not effectively managed. Conventional methods often face limitations in selectivity and efficiency, highlighting the need for innovative solutions. The P-COOH nanofiber membrane, with its high surface area and tailored functional groups, was engineered to optimize protein removal. The removal process was analyzed in a batch system at an optimal pH of 7. Various kinetic models, such as pseudo-first-order, pseudo-second-order, Avrami, and intra-particle diffusion, were applied to elucidate rate-controlling steps. In contrast, isotherm models, including Langmuir, Freundlich, and Temkin, characterized the equilibrium of the removal process. Complete elution of removed lysozyme was achieved using a 0.3 M NaCl solution, yielding an equilibrium binding capacity of 315.24 mg/g on the nanofiber membranes. These findings indicate that the Avrami nonlinear model best describes lysozyme removal's complex, multi-step kinetics. In contrast, the Langmuir linear model accurately fits the equilibrium data, suggesting monolayer removal on a homogeneous surface. The successful reusability of the membranes underscores their potential for sustainable lysozyme removal from wastewater, offering a viable solution for improved waste management in industrial applications.

本研究调查了使用弱离子交换纳米纤维膜（P-COOH）去除溶菌酶（一种常见的废弃蛋白质）的情况，以应对蛋白质废弃物管理和废水处理方面的挑战。蛋白质废物普遍存在于工业和生物过程中，如果得不到有效管理，会造成环境污染并增加处理的复杂性。传统方法往往在选择性和效率方面受到限制，因此需要创新的解决方案。P-COOH 纳米纤维膜具有高表面积和定制功能基团，可优化蛋白质去除效果。在最佳 pH 值为 7 的批处理系统中对去除过程进行了分析。各种动力学模型，如伪一阶、伪二阶、Avrami 和颗粒内扩散模型，被用于阐明速率控制步骤。相反，等温线模型，包括 Langmuir、Freundlich 和 Temkin，则描述了去除过程的平衡状态。使用 0.3 M NaCl 溶液可实现溶菌酶的完全洗脱，纳米纤维膜上的平衡结合能力为 315.24 mg/g。这些研究结果表明，阿夫拉米非线性模型最能描述溶菌酶去除的复杂、多步骤动力学。与此相反，Langmuir 线性模型精确地拟合了平衡数据，表明在均匀表面上的单层去除。膜的成功重复使用强调了其可持续去除废水中溶菌酶的潜力，为改善工业应用中的废物管理提供了可行的解决方案。

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引用次数: 0

Genosensor based on polypyrrole and dendrimer-coated gold nanoparticles for human papillomavirus detection 基于聚吡咯和树枝状聚合物包覆金纳米粒子的基因传感器，用于检测人类乳头瘤病毒

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-29 DOI: 10.1016/j.bej.2024.109551

Heloisa B. Dantas , Alberto G. Silva-Junior , Norma L.C.L. Silva , Abdelhamid Errachid , Maria D.L. Oliveira , Cesar A.S. Andrade

Human Papillomavirus (HPV) is an often asymptomatic widespread sexually transmitted infection responsible for causing various health issues. Low-risk HPV primarily causes genital warts. High-risk HPV types are associated with several cancers, including cervical, anal, and oropharyngeal cancers, posing significant health risks. In this work, we developed an electrochemical biosensor for the detection and differentiation of HPV genotypes based on electropolymerized polypyrrole (PPy) and PAMAM dendrimer-coated gold nanoparticles (PAMAM-AuNPs) for the immobilization of a DNA probe for detecting different HPV genotypes. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), and atomic force microscopy (AFM) were used to characterize the biosensor. AFM analysis revealed varied topographic surfaces associated with the increased peaks concerning the biosensor and against patient samples. Electrochemical responses indicated that the genosensor could detect HPV using plasmid (HPV 6, 16, 18, 31, and 33) and cDNA samples (HPV 6, 18, and 31) from infected patients. Different electrochemical profiles were obtained between high-risk and low-risk genotypes. The sensor presented an excellent analytical performance, presenting a lower LOD of 0.04 pg.µL⁻¹ and 0.34 pg.µL⁻¹ for plasmidial and cDNA samples, respectively. Electrochemical analysis pointed out the ability of the developed genosensor platform to differentiate the HPV genotypes. The proposed biosensor is a promising tool for detecting and monitoring HPV and related diseases such as cervical cancer.

人类乳头瘤病毒（HPV）是一种通常无症状的广泛性传播感染，可导致各种健康问题。低危型 HPV 主要导致生殖器疣。高危型 HPV 与多种癌症有关，包括宫颈癌、肛门癌和口咽癌，对健康构成重大威胁。在这项工作中，我们开发了一种用于检测和区分 HPV 基因型的电化学生物传感器，该传感器基于电聚合聚吡咯（PPy）和 PAMAM 树枝状聚合物包覆的金纳米粒子（PAMAM-AuNPs），用于固定 DNA 探针以检测不同的 HPV 基因型。电化学阻抗谱（EIS）、循环伏安法（CV）和原子力显微镜（AFM）被用来表征生物传感器。原子力显微镜分析表明，生物传感器和患者样本的峰值增加与不同的地形表面有关。电化学反应表明，该基因传感器可使用来自感染患者的质粒（HPV 6、16、18、31 和 33）和 cDNA 样品（HPV 6、18 和 31）检测 HPV。高危基因型和低危基因型的电化学特征各不相同。该传感器具有出色的分析性能，对质粒样品和 cDNA 样品的 LOD 分别为 0.04 pg.µL-1 和 0.34 pg.µL-1。电化学分析表明，所开发的基因传感器平台具有区分 HPV 基因型的能力。所提出的生物传感器是检测和监测人乳头瘤病毒及相关疾病（如宫颈癌）的一种有前途的工具。

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引用次数: 0

Self-healing of concrete crack based on modified zeolite immobilizing microorganisms 基于固定微生物的改性沸石的混凝土裂缝自愈合技术

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-28 DOI: 10.1016/j.bej.2024.109541

Nansha Ye , Zhen Liu , Peng Wang , Yongshuai Sun , Xiangli He

Selecting the appropriate carrier to safeguard the microorganisms and enhance their viability within the matrix while implementing self-healing in concrete by microbial induced calcium carbonate precipitation technology is challenging. In this study, modified zeolite was used as the microbial carrier for concrete. Varying concentrations of acid and alkali reagents were used to impregnate natural zeolite, and the optimal modification reagent was identified. The mechanical compatibility of zeolite and concrete was analyzed. The mineralization efficacy of microorganisms was assessed by analyzing the self-healing indices of concrete crack specimens, the optimal dosage of modified zeolite was determined. And the mechanism of this system was explored. The results indicate that when the bacterial solution concentration is 3×10⁷ cfu/mL, incorporating modified zeolite (particle size: 1.0–2.0 mm) with a 0.4 mol/L HCl solution yields the optimal microbial immobilization and mineralization effects. Also, with equivalent zeolite dosage, modified zeolite self-healing concrete can enhance compressive strength recovery by 8–22 % and reduce water absorption by 11–20 %, making it capable of completely repairing wider cracks. Concrete specimens with 30 % and 40 % modified zeolite content demonstrated excellent performance, with the 40 % dosage exhibiting the highest self-healing capability. At the 40 % modified zeolite dosage, concrete achieved a remarkable 86.8 % compressive strength recovery at 28 days and a 5.4 % water absorption rate, effectively repairing cracks up to 0.5 mm width.

通过微生物诱导碳酸钙沉淀技术实现混凝土自愈合的同时，选择合适的载体来保护微生物并提高其在基质中的存活率是一项挑战。在这项研究中，改性沸石被用作混凝土的微生物载体。使用不同浓度的酸碱试剂浸渍天然沸石，并确定了最佳改性试剂。分析了沸石与混凝土的机械相容性。通过分析混凝土裂缝试件的自愈合指数，评估了微生物的矿化功效，确定了改性沸石的最佳用量。并探讨了该系统的机理。结果表明，当细菌溶液浓度为 3×107 cfu/mL 时，在 0.4 mol/L HCl 溶液中加入改性沸石（粒径：1.0-2.0 mm）可获得最佳的微生物固定和矿化效果。此外，在沸石用量相等的情况下，改性沸石自修复混凝土的抗压强度恢复能力可提高 8-22%，吸水率降低 11-20%，使其能够完全修复较宽的裂缝。改性沸石含量分别为 30% 和 40% 的混凝土试样表现出优异的性能，其中 40% 的改性沸石含量表现出最高的自愈合能力。在改性沸石掺量为 40% 的情况下，混凝土在 28 天时的抗压强度恢复率高达 86.8%，吸水率为 5.4%，可有效修复宽度达 0.5 毫米的裂缝。

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引用次数: 0

CFD-guided scaling of Pseudomonas putida fermentation 以 CFD 为指导的假单胞菌发酵规模化

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-28 DOI: 10.1016/j.bej.2024.109549

Maryam Jamshidzadeh , Antonia Ursula Griesz , Jesper Wang Jensen , Ulrich Krühne , John M. Woodley , Krist V. Gernaey , Pablo Ivan Nikel , Helena Junicke

This study evaluated the scale-up of Pseudomonas putida fed-batch fermentation from a 2 L benchtop-scale stirred bioreactor to a 200 L pilot-scale tank by using a validated computational fluid dynamic (CFD) model. One of the major concerns in this fermentation process is the potential reduction in mixing quality with increasing scale, leading to lower yield or product quality. For a low-risk scale-up, a multiphase Euler-Euler CFD model was developed that simulated the hydrodynamics of the fed-batch system at various filling volumes, representing different stages of the fermentation process. The model was validated using experimental data of mixing time and mass transfer coefficient. The hydrodynamic model was then coupled with a Monod kinetic model of P. putida ‘s fermentation. Response surface methodology was used to generate a performance map of the pilot bioreactor at various aeration, agitation, and bioreactor filling volumes. The study considered different established scale-up approaches, such as constant tip speed and aeration rate across scales, constant

k_{L} a

, as well as constant power to unit of liquid volume (P/V). The performance of the bioreactor was assessed, and the optimum operating ranges of the input parameters were obtained at different stages of the fermentation. Using performance map the possibility of substrate and oxygen gradient formation, and the gradient severity inside the pilot bioreactor at different working volumes were evaluated.

本研究通过使用经过验证的计算流体动力学（CFD）模型，评估了从 2 升台式搅拌生物反应器到 200 升中试规模罐的假单胞菌给料批次发酵的放大。这种发酵工艺的一个主要问题是，随着规模的扩大，混合质量可能会降低，从而导致产量或产品质量下降。为了实现低风险放大，开发了一个多相欧拉-欧拉 CFD 模型，该模型模拟了代表发酵过程不同阶段的不同填充量下的进料批次系统的流体动力学。利用混合时间和传质系数的实验数据对模型进行了验证。然后将流体动力学模型与普氏菌发酵的莫诺动力学模型相结合。采用响应面方法生成了中试生物反应器在不同通气量、搅拌量和生物反应器填充量下的性能图。该研究考虑了不同的既定放大方法，如不同规模的恒定尖端速度和曝气速率、恒定 kLa 以及单位液体体积的恒定功率 (P/V)。对生物反应器的性能进行了评估，并得出了发酵不同阶段输入参数的最佳运行范围。利用性能图评估了基质和氧气梯度形成的可能性，以及不同工作容积下中试生物反应器内的梯度严重程度。

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引用次数: 0

Combined activated sludge and sand filtration for purification of UASB effluent with high suspended solids from water hyacinth juice 将活性污泥和砂滤结合起来，净化含有高悬浮固体的布袋莲汁污水

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biochemical Engineering Journal

Pub Date : 2024-10-24 DOI: 10.1016/j.bej.2024.109540

Akinori Fujita , Mutsumi Sekine , Masatoshi Kishi , Tatsuki Toda

Rapid biogasification of water hyacinth, a globally overgrown species, using compression and up-flow anaerobic sludge blanket (UASB) treatment of the juice has recently been proposed. This study aimed to establish a post-purification system for UASB-treated juice to effectively utilize the remaining nutrients for valuable products, such as vegetables and microalgae. To easily remove hard-to-degrade suspended solids (SS) and organic matter from UASB-treated juice, the activated sludge (AS) and sand filtration processes—conventional biological treatments—were introduced, with a focus on their physical treatment properties. Over 76 % of SS and 43 % of the organic carbon were removed from UASB-treated juice during the process. The hydraulic retention time of the AS was reduced to one day, indicating the potential of minimizing the facility size. The light transmittance, which is crucial for microalgae mediums, improved sevenfold. Even after most of the SS was removed by the AS, the sand filtration further increased the light transmittance by 1.4 times. The AS effectively removed most of the SS and organic matter, whereas sand filtration enhanced the treatment stability and further improved the light transmittance. NH₄⁺ was mostly oxidized to NO₃^-, which is less toxic to plants. The total inorganic nitrogen content remained high after treatment, suggesting that UASB effluents from water hyacinth can be used as a nutrient source for hydroponics and microalgae cultivation in developing countries.

最近，有人提出利用压缩和上流式厌氧污泥毯（UASB）处理布袋莲（一种全球过度生长的物种）汁液，对其进行快速生物气化。本研究旨在为 UASB 处理过的果汁建立一个后净化系统，以有效利用剩余的营养物质，生产蔬菜和微藻等有价值的产品。为了从 UASB 处理过的果汁中轻松去除难以降解的悬浮固体（SS）和有机物，我们引入了活性污泥（AS）和砂滤工艺--传统的生物处理方法，重点关注其物理处理特性。在此过程中，UASB 处理过的果汁中 76% 以上的 SS 和 43% 的有机碳被去除。AS 的水力停留时间缩短到了一天，这表明可以最大限度地缩小设施规模。对微藻类培养基至关重要的透光率提高了七倍。即使在自动沉淀池去除大部分 SS 后，砂滤仍将透光率进一步提高了 1.4 倍。AS 有效地去除了大部分 SS 和有机物，而砂滤增强了处理的稳定性，并进一步提高了透光率。NH4+ 大多被氧化成对植物毒性较小的 NO3-。处理后的总无机氮含量仍然很高，这表明布袋莲产生的 UASB 污水可用作发展中国家水培和微藻栽培的营养源。

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引用次数: 0