Current Research in Biotechnology最新文献

Tailored bacterial co-cultures improve Tisochrysis lutea growth and nutrient profiles under xenic conditions: a new pathway to improve microalgal production 在xenic条件下，量身定制的细菌共培养提高了黄体梭菌的生长和营养成分：一种提高微藻产量的新途径

IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-12-13 DOI: 10.1016/j.crbiot.2025.100361

Tamára F. Santos , Beatriz Simões , Veronica Rossetto , Hugo Pereira , Inês B. Maia , Marta Oliveira , Aschwin Engelen , João Navalho , João Varela

The marine haptophyte Tisochrysis lutea is a valuable source of high-value compounds, including polyunsaturated fatty acids like docosahexaenoic acid, and pigments (e.g., fucoxanthin). However, high production costs and variability remain major challenges for its large-scale application in aquaculture, pharmaceuticals, and biotechnology industries. Therefore, strategies to enhance biomass production and quality are actively explored. In natural environments, T. lutea establishes mutualistic interactions with bacteria to obtain essential nutrients such as vitamin B₁₂, yet the role of bacteria in industrial cultures remains poorly understood. In this study, 145 bacterial strains were isolated and taxonomically identified from industrial T. lutea cultures, with members of the class Gammaproteobacteria and Actinomycetia being the most prevalent. Forty isolates were screened individually in co-culture with T. lutea revealing strain-specific effects on growth and biochemical composition. Seven beneficial strains were used to design 21 tailored bacterial blends. Several consortia enhanced biomass production (up to 74 %) and increased key bioactive compounds, particularly methylcobalamin (up to 300 %). These findings demonstrate the potential of tailored bacterial consortia to enhance T. lutea productivity and nutritional quality under production-relevant xenic conditions, enabling strategic microbiome modulation for specific industrial goals.

海洋褐藻是高价值化合物的宝贵来源，包括多不饱和脂肪酸，如二十二碳六烯酸和色素（如岩藻黄素）。然而，高生产成本和可变性仍然是其在水产养殖、制药和生物技术工业中大规模应用的主要挑战。因此，积极探索提高生物质产量和质量的策略。在自然环境中，黄叶霉与细菌建立了相互作用，以获得必需的营养物质，如维生素B12，但在工业培养中，细菌的作用仍然知之甚少。在本研究中，从工业黄叶霉培养物中分离和分类鉴定了145株细菌，其中γ变形菌纲和放线菌纲的成员最为普遍。在与黄叶霉共培养的条件下，分别筛选了40株菌株，揭示了菌株对生长和生化组成的特异性影响。采用7种有益菌株设计了21种定制菌群。一些财团提高了生物质产量（高达74%），并增加了关键的生物活性化合物，特别是甲基钴胺素（高达300%）。这些发现表明，在与生产相关的xenic条件下，量身定制的细菌联合体有可能提高黄茶的生产力和营养质量，从而实现特定工业目标的战略性微生物组调节。

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引用次数: 0

CRISPR-Cas12 in immunotherapy and Beyond: Advances and challenges CRISPR-Cas12在免疫治疗及其他领域：进展与挑战

IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-12-02 DOI: 10.1016/j.crbiot.2025.100356

Tahereh Zarei Taher , Mohammad Hassan Yousefi , Hamed Afkhami , Maria Kavianpour , Amir Hossein Kheirkhah

CRISPR-Cas12 technology has greatly advanced gene editing, offering high efficiency and accuracy. Its main applications include gene modification, disease diagnosis, and especially the development of personalized cancer therapies. CRISPR-Cas12 enhances CAR-T cell therapies, allowing immune cells to target cancer more effectively. The technology also enables precise DNA sequence changes, supporting tailored treatments based on individual tumor genetics. Importantly, CRISPR-Cas12 reduces off-target effects, improving the safety and reliability of gene therapies in clinical settings. Additionally, strategies to overcome the immunosuppressive tumor microenvironment, such as modulating regulatory T cells or engineering resistance to their inhibitory signals, are emphasized. However, challenges remain, including concerns about long-term safety, delivery methods, and ethical issues related to genetic modification. Despite these hurdles, CRISPR-Cas12 offers significant potential as a safe and transformative tool for innovative immunotherapies, promising more effective cancer treatments and better patient outcomes.

CRISPR-Cas12技术极大地推进了基因编辑，具有高效率和准确性。它的主要应用包括基因修饰、疾病诊断，特别是个性化癌症治疗的发展。CRISPR-Cas12增强了CAR-T细胞疗法，使免疫细胞更有效地靶向癌症。该技术还可以实现精确的DNA序列改变，支持基于个体肿瘤遗传学的定制治疗。重要的是，CRISPR-Cas12减少了脱靶效应，提高了临床环境中基因治疗的安全性和可靠性。此外，还强调了克服免疫抑制肿瘤微环境的策略，如调节调节性T细胞或对其抑制信号的工程抗性。然而，挑战依然存在，包括对长期安全性、递送方法和与转基因相关的伦理问题的担忧。尽管存在这些障碍，CRISPR-Cas12作为创新免疫疗法的安全和变革性工具提供了巨大的潜力，有望提供更有效的癌症治疗和更好的患者预后。

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引用次数: 0

Lignocellulosic bioethanol production: a review on pretreatment strategies, biofuel separation, and artificial intelligence/machine learning − based sustainable optimization 木质纤维素生物乙醇生产：预处理策略、生物燃料分离和基于人工智能/机器学习的可持续优化研究进展

IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-11-27 DOI: 10.1016/j.crbiot.2025.100355

V. Humera Farheen , M. Aslam Abdullah , I. Ganesh Moorthy

Bioethanol production from lignocellulosic biomass, such as agricultural residues, forestry waste, and energy crops, is an abundant, renewable, and non-food-competing resource that has gained attention as a sustainable alternative to fossil fuels. The conversion process typically involves three stages: pretreatment, enzymatic hydrolysis, and fermentation. Pretreatment using physical, chemical, biological, and nanotechnological-assisted methods is essential to break down the rigid biomass structure and enhance enzyme accessibility. In the subsequent enzymatic hydrolysis, cellulolytic enzymes convert cellulose and hemicellulose into fermentable sugars. Fermentation, carried out by microorganisms such as Saccharomyces cerevisiae, Zymomonas mobilis, or Pichia stipitis, subsequently convert these sugars into ethanol. Despite its potential, lignocellulosic bioethanol production faces challenges such as high pretreatment costs, enzyme inefficiency, and the presence of fermentation inhibitors. The use of statistical and optimization tools such as Artificial Neural Networks (ANN) and Response Surface Methodology (RSM) has been widely used to enhance bioethanol production. ANN provides robust predictive modelling by handling complex, non-linear interactions, while RSM enables efficient experimental design, identification of factor interactions, and optimization with fewer trials. Combining these methods improves ethanol yield, minimizes inhibitors, and enhances process efficiency. Recent advancements in metabolic engineering, microbial strain development, and integrated bioprocessing approaches have contributed to improving ethanol yield and process efficiency. This review explores recent progress in bioethanol production from lignocellulosic biomass, focusing on technological innovations, challenges, and future research directions aimed at enhancing the economic viability of lignocellulosic bioethanol as a renewable fuel source.

从木质纤维素生物质（如农业残留物、林业废弃物和能源作物）中生产生物乙醇是一种丰富的、可再生的、不与食物竞争的资源，作为化石燃料的可持续替代品而受到关注。转化过程通常包括三个阶段：预处理、酶解和发酵。使用物理、化学、生物和纳米技术辅助的预处理方法对于打破刚性生物质结构和提高酶的可及性至关重要。在随后的酶解过程中，纤维素水解酶将纤维素和半纤维素转化为可发酵的糖。发酵由酿酒酵母菌、活动单胞菌或毕赤酵母等微生物进行，随后将这些糖转化为乙醇。尽管具有潜力，但木质纤维素生物乙醇生产面临着预处理成本高、酶效率低和发酵抑制剂存在等挑战。利用统计和优化工具，如人工神经网络（ANN）和响应面法（RSM）已被广泛用于提高生物乙醇的生产。人工神经网络通过处理复杂的非线性相互作用提供了强大的预测建模，而RSM能够有效地进行实验设计，识别因素相互作用，并通过更少的试验进行优化。结合这些方法可以提高乙醇产量，减少抑制剂，提高工艺效率。代谢工程、微生物菌株开发和综合生物处理方法的最新进展有助于提高乙醇产量和工艺效率。本文综述了木质纤维素生物质生产生物乙醇的最新进展，重点介绍了技术创新、挑战和未来的研究方向，旨在提高木质纤维素生物乙醇作为可再生燃料来源的经济可行性。

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引用次数: 0

Plant vs mammal extracellular vesicles: new tools in therapeutic drug delivery 植物与哺乳动物的细胞外囊泡：治疗药物输送的新工具

IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-11-21 DOI: 10.1016/j.crbiot.2025.100352

Elisa Maricchiolo, Pasquale Creanza, Michela Osnato, Mattia Tiboni, Luca Casettari, Annalisa Aluigi, Andrea Pompa

Extracellular vesicles (EVs) are highly promising biological carriers, due to their potential for drug delivery and their involvement in multiple cellular processes. These vesicles, released by cells into the extracellular space, can be classified into different types based on their origin, size, and molecular content. While animal-derived EVs have been extensively studied and characterised, recent research has highlighted the growing interest in plant-derived EVs, which present unique properties that could offer novel advantages in therapeutic applications.

This review aims to provide an overview of similarities and differences between animal and plant-derived EVs, focusing on their structural and functional characteristics. Additionally, we report various methods used for the isolation and characterisation of these vesicles, to provide a comprehensive summary, highlighting the main challenges and opportunities of each experimental option.

Moreover, the present work explores the most used techniques for loading bioactive molecules into EVs, transforming them from natural biological entities into innovative drug delivery systems. The ability to encapsulate drugs, small RNA molecules, or proteins within EVs opens up new frontiers for the targeted treatment of diseases, with particular emphasis on overcoming barriers related to drug bioavailability and specificity.

Finally, this review stresses the importance of ongoing research in the extracellular vesicles field, from both animal and plant origins, given their adaptable properties to multiple pharmaceutical purposes. Plant-derived EVsoffer a sustainable and economic alternative, together with the better characterised mammalian vesicles, showing the potential to strongly improve the drug delivery strategy in the near future.

细胞外囊泡（EVs）是非常有前途的生物载体，因为它们具有潜在的药物递送和参与多种细胞过程。这些囊泡由细胞释放到细胞外空间，根据它们的来源、大小和分子含量可分为不同类型。虽然动物源性电动汽车已经被广泛研究和表征，但最近的研究强调了对植物源性电动汽车日益增长的兴趣，因为植物源性电动汽车具有独特的特性，可以在治疗应用中提供新的优势。本文综述了动物源性电动汽车和植物源性电动汽车的异同，重点介绍了它们的结构和功能特征。此外，我们报告了用于分离和表征这些囊泡的各种方法，以提供一个全面的总结，突出每个实验选项的主要挑战和机遇。此外，本研究还探索了将生物活性分子装载到电动汽车中，将其从天然生物实体转化为创新的药物输送系统的最常用技术。在ev内封装药物、小RNA分子或蛋白质的能力为疾病的靶向治疗开辟了新的领域，特别强调克服与药物生物利用度和特异性相关的障碍。最后，本综述强调了细胞外囊泡领域正在进行的研究的重要性，从动物和植物的起源，考虑到它们对多种药物用途的适应性。植物源性ev软膏是一种可持续和经济的替代品，与更好表征的哺乳动物囊泡一起，显示出在不久的将来强有力地改善药物递送策略的潜力。

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引用次数: 0

The landscape of LncRNAs in diabetic kidney disease: a meta-analysis of transcriptomics data lncrna在糖尿病肾病中的作用：转录组学数据的荟萃分析

IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-07-30 DOI: 10.1016/j.crbiot.2025.100322

Raziyeh Rezaei , Basireh Bahrami , Yousof Gheisari

Non-coding regions of the genome are known to influence complex disorders, yet the role of long non-coding RNAs (lncRNAs) in Diabetic Kidney Disease (DKD) remains underexplored. This study conducts a meta-analysis of RNA-sequencing data from murine kidney samples of type 1 (T1DM) and type 2 diabetes mellitus (T2DM) to identify lncRNAs associated with DKD. DKD-associated datasets were harvested, and after data pre-processing and quality assessment, 6 T1DM and 4 T2DM datasets were included. Data integration, batch correction, and normalization were performed, followed by the identification of differentially expressed lncRNAs (meta-DELs) and mRNAs (meta-DEMs). A DKD mouse model was developed to validate the expression of selected meta-DELs using qRT-PCR. The meta-analysis identified 188 meta-DELs in T1DM and 68 in T2DM. Notably, a small set of lncRNAs have dense mRNA interactions, including Dancer, Gm7628, C4a, and Gm17300 in T1DM and Malat1, C4a, Gm17300, and Eif4a2 in T2DM. Expression analysis confirmed the up-regulation of seven selected meta-DELs in the DKD model, with Trp53cor1, Gm15462, and Gm42664 reaching statistical significance. This systematic analysis of high-quality expression profiles identified meta-DELs consistently associated with DKD, distinguishing actual lncRNA changes from those influenced by experimental conditions or gene expression noise.

众所周知，基因组的非编码区会影响复杂的疾病，但长链非编码rna （lncRNAs）在糖尿病肾病（DKD）中的作用仍未得到充分探索。本研究对1型（T1DM）和2型糖尿病（T2DM）小鼠肾脏样本的rna测序数据进行了荟萃分析，以确定与DKD相关的lncrna。收集与dkd相关的数据集，经过数据预处理和质量评估，包括6个T1DM和4个T2DM数据集。进行数据整合、批量校正和归一化，随后鉴定差异表达的lncrna （meta-DELs）和mrna （meta- dem）。建立DKD小鼠模型，利用qRT-PCR验证所选meta-DELs的表达。荟萃分析发现，T1DM患者有188例，T2DM患者有68例。值得注意的是，一小部分lncrna具有密集的mRNA相互作用，包括T1DM中的Dancer、Gm7628、C4a和Gm17300，以及T2DM中的Malat1、C4a、Gm17300和Eif4a2。表达分析证实了DKD模型中7个选定的meta-DELs的上调，其中Trp53cor1、Gm15462和Gm42664达到了统计学意义。这项高质量表达谱的系统分析发现了与DKD一致相关的meta- del，将实际的lncRNA变化与受实验条件或基因表达噪声影响的lncRNA变化区分开来。

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引用次数: 0

Gut health improvement by locally isolated probiotics and histomorphometric analysis in Wistar rats 局部分离益生菌改善Wistar大鼠肠道健康及组织形态计量学分析

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-01-01 DOI: 10.1016/j.crbiot.2024.100271

Zuhra Bibi , Dilara Abbas Bukhari , Muhammad Qadeer Sarwar , Arifullah , Samina Younas , Tayyab Manzoor , Abdul Rehman

In the present investigation, lab-isolated probiotics Weisella confusa MZ735961.1, Lactiplantibacillus plantarum MZ707748.1, L. plantarum MZ710117.1, and L. plantarum MZ735961 were used separately and in combinations to evaluate their effect on gut morphology of Wistar rats. Synergistic groups were formed by 1:1 and labeled as G1 (L. plantarum MZ707748.1 and L. plantarum MZ729681.1), G2 (W. confusa MZ735961.1 and L. plantarum MZ727611.1), G3 (L. plantarum MZ729681.1, W. confusa MZ735961.1, and Lactobacillus acidophilus La-14), G4 (all above mentioned probiotics). Rats were gavage-fed with probiotics according to their colony-forming unit (CFU). The experiment was carried out for 35 days. The bacteria were re-isolated from the gut and identified by biochemical tests which confirmed the administration and re-isolation of different Lactobacillus strains from the gut. Molecular characterization was done through 16S rRNA by using universal primers. After sequencing eight Lactobacillus strains were identified. Histopathology of rats’ intestines was done, and different parameters were examined. Villus height, crypt height, crypt width, mucosa, and sub-mucosa of jejunum were significantly (p = 0.00) increased in the G3 synergetic probiotic group compared to 0-day and negative control. However, the villus width showed non-significant (p > 0.05) variations in both genders. Mucosa tunic, muscle tunic, total wall, and crypt depth were significantly increased (p = 0.00) in the G4 group of medial colon. The study concluded that gut morphology improves as probiotics adhere better to the intestinal epithelium, excluding pathogens, reducing inflammation, enhancing nutrient absorption, and stimulating mucosal growth. This results in improved villus structure and gut wall integrity.

本研究采用实验室分离的几种益生菌，分别使用杂交魏氏菌MZ735961.1、植物乳杆菌MZ707748.1、植物乳杆菌MZ710117.1和植物乳杆菌MZ735961，观察其对Wistar大鼠肠道形态的影响。按1:1组成协同组，分别标记为G1 （L. plantarum MZ707748.1和L. plantarum MZ729681.1）、G2 （W. confusa MZ735961.1和L. plantarum MZ727611.1）、G3 （L. plantarum MZ729681.1、W. confusa MZ735961.1和嗜酸乳杆菌La-14）、G4（上述益生菌）。大鼠按菌落形成单位（CFU）灌喂益生菌。试验期35 d。从肠道中重新分离细菌，并通过生化试验鉴定，证实了肠道中不同乳杆菌菌株的给药和重新分离。使用通用引物通过16S rRNA进行分子鉴定。测序后鉴定出8株乳杆菌。对大鼠肠道进行组织病理学检查，并对各参数进行检测。与0日龄和阴性对照组相比，G3协同益生菌组空肠绒毛高度、隐窝高度、隐窝宽度、黏膜和亚黏膜显著（p = 0.00）升高。但绒毛宽度无显著差异(p >；0.05)的差异。G4组内结肠粘膜、肌被膜、总肠壁、隐窝深度均显著增加（p = 0.00）。该研究得出结论，肠道形态改善，因为益生菌更好地粘附在肠上皮上，排除病原体，减少炎症，促进营养吸收，刺激粘膜生长。这改善了绒毛结构和肠壁的完整性。

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引用次数: 0

Corrigendum to “Postbiotics, a natural feed additive for growth performance, gut microbiota and quality of poultry products” [Curr. Res. Biotechnol. 8 (2024) 100247] 更正："益生菌，一种促进家禽生长性能、肠道微生物群和产品质量的天然饲料添加剂"[Curr. Res. Biotechnol.

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-01-01 DOI: 10.1016/j.crbiot.2025.100284

Jakub Urban , Karwan Yaseen Kareem , Atanas G. Atanasov , Arkadiusz Matuszewski , Damian Bień , Patrycja Ciborowska , Anna Rygało-Galewska , Monika Michalczuk

引用次数: 0

Validation of microbial source tracking markers through PCR-based molecular analysis and microbial genome database 通过基于pcr的分子分析和微生物基因组数据库验证微生物源跟踪标记

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-01-01 DOI: 10.1016/j.crbiot.2025.100304

Rifat Zubair Ahmed , Ashraful Islam , Tiantian Tian , Zhe Tian , Yu Zhang , Min Yang

Microbial Source Tracking (MST) uses molecular markers targeting host-associated gut microorganisms to identify fecal pollution. However, MST faces significant challenges in fecal source identification, particularly due to the markers’ poor specificity and shared genomic areas among microorganisms from different host sources. This study addresses these challenges by using host-specific Escherichia coli genetic markers, originally developed through a novel, library-independent approach, to detect sources of fecal pollution. A total of 563 E.coli isolates from chicken, cow, and pig feces were isolated and assessed by nine reported host-associated E. coli genetic markers (Chicken: CH7, CH9, CH12, CH13; Cow: CO2, CO3; Pig: P1, P3, P4) through PCR. Marker possession patterns, sensitivity, specificity, and accuracy were calculated. The NCBI Microbial Genome database was searched for sequences homologous to genome regions of studied genetic markers and evaluated by finding the percentage of host sources and sequence location in the genome. Homology evaluation with binary PCR results was used to predict the best-performing marker. PCR results exhibited that the most effective markers were chicken CH7 (67% sensitivity, 77.9% specificity, 74.4% accuracy) and CH9 (55% sensitivity, 99.4% specificity, 84.7% accuracy). However, a homology search in the database narrowed the selection of the top-performing marker to CH7, which showed homology with E.coli from chicken hosts, while other markers exhibited higher homology with E.coli from Humans. Furthermore, sequences from the database homologous to the CH9 and CO2 markers were found on a plasmid, while those for CH12, CO3, P1, and P4 were on the chromosome, and CH7, CH13, and P3 were on both. This study highlights the critical need for integrated approaches to assess molecular markers in MST assays, emphasizing their significance in advancing research within the field.

微生物源追踪技术（MST）是一种利用分子标记靶向宿主相关肠道微生物来识别粪便污染的技术。然而，MST在粪便来源鉴定方面面临着重大挑战，特别是由于标记的特异性较差，并且来自不同宿主来源的微生物具有相同的基因组区域。本研究通过使用宿主特异性大肠杆菌遗传标记来解决这些挑战，该标记最初是通过一种新颖的、独立于文库的方法开发的，用于检测粪便污染源。从鸡、牛和猪粪便中分离出563株大肠杆菌，并利用9个已报道的宿主相关大肠杆菌遗传标记(鸡：CH7、CH9、CH12、CH13；牛：CO2， CO3；猪：P1， P3, P4)。计算标记物占有模式、敏感性、特异性和准确性。在NCBI微生物基因组数据库中搜索与所研究遗传标记的基因组区域同源的序列，并通过寻找宿主来源的百分比和序列在基因组中的位置进行评估。用二元PCR结果进行同源性评价，预测最佳标记。PCR结果显示，鸡CH7（敏感性67%，特异性77.9%，准确性74.4%）和CH9（敏感性55%，特异性99.4%，准确性84.7%）是最有效的标记。然而，在数据库中进行同源性搜索，将选择的最佳标记缩小到CH7，该标记与鸡宿主大肠杆菌具有同源性，而其他标记与人类大肠杆菌具有较高的同源性。此外，在一个质粒上发现了数据库中与CH9和CO2标记同源的序列，而CH12、CO3、P1和P4标记在染色体上，CH7、CH13和P3标记在两条染色体上。这项研究强调了在MST检测中评估分子标记的综合方法的迫切需要，强调了它们在推进该领域研究中的重要性。

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引用次数: 0

Cost-effective production process of scFv antibody fragments against Shiga toxin 2 via recombinant E. coli 利用重组大肠杆菌生产抗志贺毒素2单链病毒抗体片段的成本效益研究

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-01-01 DOI: 10.1016/j.crbiot.2025.100310

Marcela Guimarães , Daniela Luz , Elisabeth de Fátima Pires Augusto , Lucia Vieira , Maricilia Silva Costa , Roxane Maria Fontes Piazza , José Geraldo da Cruz Pradella

Shiga toxin (Stx)-producing Escherichia coli (STEC) and its subgroup enterohemorrhagic E. coli are significant pathogens responsible for diarrhea, which can progress to hemorrhagic colitis and hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children. Early diagnosis is crucial for effective clinical management, as antibiotic treatment is not recommended for STEC infections. The present study aimed to establish a cost-effective biotechnological platform for cultivating recombinant E. coli to produce scFv antibody fragments against Stx2 for diagnostic applications. The method was first evaluated through shake flask experiments and subsequently scaled up to bench-scale bioreactors operated in both batch and fed-batch modes using defined culture media. Optimal production conditions were achieved by inducing recombinant E. coli pLys at 18 °C for 18 h with 0.1 mM IPTG, resulting in a yield of 3.0 to 4.0 mg scFv/g cell biomass. A fed-batch, high-cell-density procedure with E. coli pLysS achieved a maximum production up to 150 mg scFv/L. A preliminary economic assessment demonstrated the production potential at a value of around $250/g scFv. Economic analysis also highlights that the relative cost of capital investment becomes important as production processes intensify. Therefore, technical parameters such as productivity (scFv mass/bioreactor volume * time) and scFv concentration (mass scFv mass/bioreactor volume) should be prioritized to maximize their values. Similarly, optimization of the recombinant E. coli microbial platform should be pursued to increase the Yp/x level.

产志贺毒素（Stx）的大肠杆菌（STEC）及其亚群肠出血性大肠杆菌是导致腹泻的重要病原体，可发展为出血性结肠炎和溶血性尿毒症综合征（HUS），是儿童急性肾衰竭的主要原因。早期诊断对于有效的临床管理至关重要，因为不建议对产志毒素大肠杆菌感染进行抗生素治疗。本研究旨在建立一个具有成本效益的培养重组大肠杆菌的生物技术平台，以生产用于诊断的Stx2的scFv抗体片段。该方法首先通过摇瓶实验进行评估，随后扩展到使用确定的培养基在批处理和补料批处理模式下运行的实验规模生物反应器。最佳的生产条件是在18°C下，用0.1 mM IPTG诱导重组大肠杆菌pLys 18 h，产量为3.0 ~ 4.0 mg scFv/g细胞生物量。使用大肠杆菌pLysS进行分批喂料、高密度处理，最大产量可达150 mg scFv/L。初步经济评估表明，其生产潜力约为250美元/克scFv。经济分析还强调，随着生产过程的加剧，资本投资的相对成本变得越来越重要。因此，应优先考虑生产效率（scFv质量/生物反应器体积*时间）和scFv浓度（质量scFv质量/生物反应器体积）等技术参数，使其值最大化。同样，重组大肠杆菌的微生物平台也需要优化，以提高Yp/x水平。

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引用次数: 0

Bioremediation of heavy metals from electronic waste dumping sites with bacteria 利用细菌对电子废弃物倾倒场重金属进行生物修复

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Current Research in Biotechnology

Pub Date : 2025-01-01 DOI: 10.1016/j.crbiot.2025.100309

Husnain Ahmad Khan , Shahid Sher , Dilara Abbas Bukhari , Abdul Rehman

Samples were collected from two e-waste dumping sites (Mehmood Booti (31°36′28″N, 74°23′36″E) and Lakhodair (31°37′36.6″ N, 74°25′07.6″ E)) in Lahore, Pakistan. A portable multiparameter was used to determine physicochemical parameters such as temperature, pH, electrical conductivity, turbidity, total suspended particles, and total dissolved solids. Minimal salt broth was used for the determination of the minimal inhibitory concentration of the bacterium against all heavy metals. Bacterial morphology was observed under a scanning electron microscope with and without metal stress. The temperature range for all these samples was 28.7 to 35.7 °C, while the pH range was 6.7 to 7.89. The other parameters range, such as electrical conductivity µS/cm (698–8742), turbidity (14.2–103), total suspended particles (31–698), and total dissolved solids (564–23456). The lead concentration in the Mehmood Booti soil sample was 1800 mg/kg, while in the Lakhodair soil, it was 1567 mg/kg. Microbacterium sp. strain 1S1 was utilized for bioremediation assay at the lab and pilot scale. The resistance capacity of this bacterium against different metals was in the following order: As > Pb > Cd > Cu > Cr > Ni. The bioremediation potential of the bacterium against arsenic was 81.33 % and 96 % after 2 and 4 days. The least activity was observed against nickel, which was 17 and 28.33 % after 2 and 4 days. The metal removal capacity per CFU was the maximum for lead and arsenic compared to other metals, which were 1.99E-7 and 1.45E-07. The heat-inactivated bacterial cells removed arsenic in higher concentrations and lead in lower concentrations. The electron microscopy showed no significant alteration in bacterial morphology in control and metal-treated bacterial cells. The nanopore long-read sequencing analysis revealed that cadmium, nickel, copper, and arsenic resistance genes were found on the bacterial genome. No genes were found for lead and chromium but 849 hypothetical coding sequences having unknown functions were present on the bacterial genome. So, the Microbacterium sp. strain 1S1 is a potential candidate for the removal of heavy metals from e-waste dumping sites.

样本采集自巴基斯坦拉合尔的两个电子垃圾倾倒场（Mehmood Booti（31°36′28″N, 74°23′36″E）和Lakhodair(31°37′36.6″N, 74°25′07.6″E)）。便携式多参数仪用于测定理化参数，如温度、pH、电导率、浊度、总悬浮颗粒和总溶解固体。用微量盐肉汤测定细菌对所有重金属的最低抑菌浓度。在扫描电子显微镜下观察有无金属应力的细菌形态。所有样品的温度范围为28.7 ~ 35.7℃，pH范围为6.7 ~ 7.89。其他参数范围，如电导率µS/cm(698-8742)，浊度（14.2-103），总悬浮颗粒（31-698）和总溶解固体（564-23456）。Mehmood Booti土壤样品的铅浓度为1800 mg/kg， Lakhodair土壤样品的铅浓度为1567 mg/kg。在实验室和中试规模上采用微杆菌1S1菌株进行生物修复试验。该细菌对不同金属的抗性强弱顺序为：As >；Pb祝辞Cd比;铜比;Cr祝辞倪。2 d和4 d后，细菌对砷的生物修复率分别为81.33%和96%。2 d和4 d对镍的活性最低，分别为17%和28.33%。每CFU对铅和砷的金属去除率最高，分别为1.99E-7和1.45E-07。热灭活的细菌细胞去除高浓度的砷和低浓度的铅。电镜观察显示，对照组和金属处理过的细菌细胞形态无明显变化。纳米孔长读测序分析显示，在细菌基因组上发现了抗镉、抗镍、抗铜和抗砷基因。没有发现铅和铬的基因，但在细菌基因组中存在849个功能未知的假设编码序列。因此，微细菌sp.菌株1S1是去除电子垃圾倾倒场重金属的潜在候选者。

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