Current Research in Biotechnology最新文献

{"title":"Characterization of a novel thermostable NAD+-dependent formate dehydrogenase from Methylacidiphilum kamchatkense Kam1 (MkaFDH)","authors":"Khouloud Zribi ,&nbsp;Matteo Ciciani ,&nbsp;Agata Sofia Assunção Carreira ,&nbsp;Martina Paganin ,&nbsp;Sara Pozzo ,&nbsp;Lucio Cinà ,&nbsp;Baris Binay ,&nbsp;Francesco Secundo ,&nbsp;Nicola Segata ,&nbsp;Alessandro Provenzani","doi":"10.1016/j.crbiot.2025.100306","DOIUrl":"10.1016/j.crbiot.2025.100306","url":null,"abstract":"<div><div>Metal-independent NAD⁺-dependent formate dehydrogenases (FDHs) are enzymes responsible for catalyzing the conversion of formate (HCOO^–) to carbon dioxide (CO₂), a biological reaction involved in microbial carbon processing and cofactor regeneration. These enzymes show large potential for environmental bioremediation and biotechnological uses. However, FDHs applications are hampered by the enzymes’ limited stability under extreme conditions, such as high temperatures or extreme pH. Therefore, we aimed to identify and characterize novel metal-independent FDHs with improved activity and thermostability compared to known FDHs. By using four different FDH protein sequences, <em>Ct</em>FDH (from <em>Chaetomium thermophilum)</em>, <em>Mt</em>FDH (from <em>Myceliophthora thermophile)</em>, <em>Op</em>FDH (from <em>Ogata parapolymorpha</em> DL-1) and <em>Pse</em>FDH (from <em>Pseudomonas</em> sp.<em>101)</em> we retrieved 18,850 FDHs sequences from the NCBI database and matched against the species present in the database of thermophilic bacteria, ThermoBase. Our phylogenetic analysis identified four distinct FDHs in thermophilic bacteria: <em>Methylocaldum szegediense</em> (<em>Msz</em>FDH), <em>Methylacidiphilum kamchatkense</em> (<em>Mka</em>FDH), <em>Mycobacterium arosiense</em> (<em>Mar</em>FDH) and <em>Mycobacterium genavense</em> (<em>Mge</em>FDH). We selected and characterized the <em>Mka</em>FDH as it was expressed in the thermophilic bacterium with the highest optimum growth (55 °C) among the four bacteria. The <em>MkaFDH</em> was cloned, and the recombinant protein was expressed in <em>E. coli</em> and purified. The conditions for the optimal catalytic activity for formate oxidations were screened and identified, revealing metal-independent, NAD⁺-restricted activity in phosphate buffer, pH 8. Importantly, the enzyme showed remarkable thermal stability and catalytic activity, showing a melting temperature (Tm) of 60.15 °C, as confirmed by far-UV circular dichroism (CD). Finally, the enzyme showed good thermostability for formate oxidation up to 57.5 °C, and its high catalytic efficiency (k_cat/K_m = 0.44 s⁻¹mM⁻¹) suggested its potential industrial application. Collectively, we describe here a novel FDH with relevant thermostability that can be exploited as a prototype for industrial applications.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100306"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144366626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of artificial intelligence in automating diagnostic procedures in clinical microbiology laboratories","authors":"Nishant Singhal ,&nbsp;Harsh Vardhan ,&nbsp;Rajul Jain ,&nbsp;Payal Gupta ,&nbsp;Aaysha Pandey ,&nbsp;Naresh Kumar Wagri ,&nbsp;Ashish Gaur","doi":"10.1016/j.crbiot.2025.100351","DOIUrl":"10.1016/j.crbiot.2025.100351","url":null,"abstract":"<div><div>With infectious diseases continuing to pose a significant challenge to global health, clinical laboratories are pursuing faster, more accurate, and more scalable diagnostic options. This article highlights how advancements in robotics, machine learning, deep learning, and natural language processing are revolutionizing traditional laboratory practices. From automated Gram-staining and slide analysis to AI-enabled bacterial identification and antibiotic resistance testing, every technological development enhances diagnostic precision, reduces human error, and speeds up turnaround times. The assessment also deals with the real-world challenges of integrating these technologies, which include ethical issues, data privacy, system compatibility, and user acceptance. Additionally, it examines possible future developments, such as rapid diagnostics, smart laboratory infrastructure, and AI’s capability to create a seamless, interconnected network of diagnostic tools. As laboratories move towards completely automated and intelligent systems, combining human expertise with machine intelligence may enhance microbiological diagnostics’ quality, efficiency, and responsiveness in clinical settings.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100351"},"PeriodicalIF":4.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145570910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel methyl-substituted pyrrole hydrazones as selective melanoma agents","authors":"Stanislava Vladimirova ,&nbsp;Rossitsa Hristova ,&nbsp;Ivan Iliev","doi":"10.1016/j.crbiot.2025.100354","DOIUrl":"10.1016/j.crbiot.2025.100354","url":null,"abstract":"<div><div>Melanoma represents the most aggressive form of skin cancer, derived from melanocytes − the pigment-producing cells located in the epidermis. Effective treatment of melanoma remains a major clinical challenge, largely due to its capacity for immune evasion, rapid metastatic spread, and the development of resistance to therapeutic interventions. Based on our previously developed compounds with high selectivity towards melanoma SH-4 cell line, we created a new series of compounds in which we replaced the halogen atom with a methyl group in the precursor pyrrole-based carbohydrazide <strong>2</strong>. All derivative hydrazones (<strong>2A–D</strong>) were synthesized by the condensation of carbohydrazide <strong>2</strong> with various substituted pyrrole aldehydes, subsequently characterized, and subjected to detailed spectroscopic analysis. <em>In vitro</em> evaluation of these newly developed compounds was performed across a diverse set of cancerous and non-cancerous cell lines to determine their biological effects. A safety profile was established using the BALB 3 T3 NRU-assay with mouse embryonic fibroblasts (BALB 3 T3 clone A31), revealing minimal cytotoxicity and an absence of phototoxic effects. Antiproliferative effects were quantified in keratinocytes (HaCaT) and melanoma (SH-4) cells using the MTT dye reduction assay. Of particular note, compound <strong>2C</strong> emerged as the most selective agent (SI = 5.51) against human melanoma cells, displaying significant antiproliferative activity (IC₅₀ = 31.93 ± 2.59). Further investigation revealed that the cytotoxic action of <strong>2C</strong> is mediated by the induction of cell accumulation in the S/G2 phase. Consistent with our prior research, the current results further confirm that hydrazones synthesized via condensation with β-aldehydes exhibit enhanced bioactivity relative to those obtained from α-aldehydes.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100354"},"PeriodicalIF":4.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145617415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “Lipid droplets dependent or independent cytoprotective activities of unsaturated fatty acids, Lorenzo’s oil and sulfo-N-succinimidyl oleate on 7-ketocholesterol-induced oxidative stress, organelle dysfunction and cell death on 158N and ARPE-19 cells: Cell targets and benefits of sulfo-N-succinimidyl oleate” [Curr. Res. Biotechnol. 7 (2024) 100195]","authors":"Thomas Nury ,&nbsp;Imen Ghzaiel ,&nbsp;Aziz Hichami ,&nbsp;Claudio Caccia ,&nbsp;Valerio Leoni ,&nbsp;Vivien Pires ,&nbsp;Atanas G Atanasov ,&nbsp;Amira Zarrouk ,&nbsp;Gérard Lizard ,&nbsp;Anne Vejux","doi":"10.1016/j.crbiot.2025.100285","DOIUrl":"10.1016/j.crbiot.2025.100285","url":null,"abstract":"","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"9 ","pages":"Article 100285"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143834804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “A review: Anti-obesity drug discovery from natural plant metabolites and endogenous peptides” [Curr. Res. Biotechnol. 8 (2024) 100259]","authors":"Xiaomu Zhu ,&nbsp;Dongdong Wang ,&nbsp;Atanas G. Atanasov","doi":"10.1016/j.crbiot.2025.100289","DOIUrl":"10.1016/j.crbiot.2025.100289","url":null,"abstract":"","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"9 ","pages":"Article 100289"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143834803","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Workflow for evaluating enzyme immobilization and performance for continuous flow manufacturing","authors":"Emily M. Luteran,&nbsp;Marc R. Aloisi,&nbsp;Wendi L. Akerley,&nbsp;Robert D. Gilbertson","doi":"10.1016/j.crbiot.2025.100327","DOIUrl":"10.1016/j.crbiot.2025.100327","url":null,"abstract":"<div><div>Enzymes have shown promise in various industries due to their functional specificity, catalytic efficiency, and environmental sustainability. These biological catalysts can be a pivotal component of manufacturing pipelines like continuous flow chemistry. For this, there exists a need to robustly immobilize enzymes on solid supports and assess the effects of the solid supports on catalytic performance and stability. Here, we use an industrially relevant model enzyme, <em>C. ensiformis</em> (Jack bean) urease, to demonstrate immobilization and assess performance in the context of continuous flow manufacturing. Various immobilization strategies were screened focusing on immobilization efficiency, protocol simplicity, and urease biocatalyst kinetics. Based on this, CDI-agarose and NHS-agarose resins were identified as the best-performing immobilization strategies for urease. CDI-agarose-urease and NHS-agarose-urease were then scaled up and applied to a large-scale continuous flow reactor to evaluate product yields, operational stability, and long-term stability. These experiments identified differences in stability and performance depending on the immobilization method tested. This highlights the importance of screening immobilization methods and subsequent enzyme performance for each candidate biocatalyst used in manufacturing to promote optimal performance and stability. As such, this work provides a framework for evaluating enzyme biocatalyst immobilization approaches to improve performance and enable transition into industrial processes.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100327"},"PeriodicalIF":4.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144866518","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and biological study of novel FELL analogs containing L- or d-tyr instead of l-phe in the N-terminus","authors":"Boryana Borisova ,&nbsp;Hristina Nocheva ,&nbsp;Hristina Zlatanova-Tenisheva ,&nbsp;Marie Laronze-Cochard ,&nbsp;Stéphane Gérard ,&nbsp;Dancho Danalev","doi":"10.1016/j.crbiot.2025.100332","DOIUrl":"10.1016/j.crbiot.2025.100332","url":null,"abstract":"<div><div>The high prevalence of pain affecting millions of people worldwide made it a major health problem. At the same time often, inflammation is closely associated to the pain. Thus, creation of molecules with a double effect is a good alternative to the currently existing in the medicinal practice non-steroidal medications. Herein, some modifications in the N- and C-terminus of the tetrapeptide FELL with proven anti-inflammatory properties are performed and the newly synthesized compounds are tested for both analgesic and anti-inflammatory potential. The analgesic and anti-inflammatory activity of the newly synthesized molecules was investigated using Paw-pressure and Carrageenan-Induced Paw Edema tests, respectively, on experimental animals. The results showed a certain “discrepancy” between the analgesic and the anti-inflammatory effects of the newly synthesized substances. Newly synthesized <strong>BB9</strong> and <strong>BB15</strong>, L-Tyr containing C-terminal amide and free acid, respectively, exhibit a more significant analgesic activity compared to those of parent molecules <strong>BB1</strong> and <strong>BB11</strong>, containing L-Phe in the N-terminus. Moreover, taking into account the obtained results for the compounds <strong>BB10</strong> and <strong>BB16</strong>, the preferable substitution is L-Tyr, instead of D-Tyr. It could be concluded that the aromatic OH-function is probably important for the connection with the opioid and cannabinoid receptors. However, the designed structural modifications did not lead to improvement in anti-inflammatory effect compared to those of parent molecules. Moreover, in a context of the positive finding is that all modifications done save the hydrolytic stability of the molecules.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100332"},"PeriodicalIF":4.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144922612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dark interruption with red or blue LEDs mitigates powdery mildew and enhances bioactive compound accumulation in organically grown strawberries","authors":"Samar Khalil ,&nbsp;Beatrix W. Alsanius ,&nbsp;Most Tahera Naznin","doi":"10.1016/j.crbiot.2025.100313","DOIUrl":"10.1016/j.crbiot.2025.100313","url":null,"abstract":"<div><div>Disease attack of powdery mildew (Podosphaera aphanis) is a major concern in organic strawberry production in tunnels or greenhouses production. We examined the impact of dark period interruption with light from blue and red LEDs on the occurrence of powdery mildew in organically grown strawberries, plant performance and on berry quality. Strawberry cultivars (Fragaria × ananassa cvs. Honeye and Faith) were grown in a climate chamber for two months in pots filled with peat based growing media certified for organic production. The plants were drip irrigated and fertilized with liquid organic fertilizers. They were exposed to a control treatment with 18 h light using white (polychromatic) LEDs and 6 h darkness or with treatments involving white LED treatment for 14 h, followed by darkness, interrupted after 2 h by 4 h of monochromatic blue or red LED exposure and additional 4 h of darkness. A three-factorial experiment including strawberry cultivars, light regimes and pathogen inoculation was performed using six replicates (pots) per cultivar and treatment. Light regime influenced the biomass of strawberry plants irrespective of cultivar, and dark period interruption with using red or blue LEDs promoted fresh biomass of the canopy and roots as compared to the control regime. Dark interruption using blue LEDs enhanced the accumulated berry yield, increased the antioxidant activities and reduced the disease incidence as compared to the control regime and the dark period interruption using red LEDs. Blue LED treatment favored the performance of the strawberry cultivars. The obtained results are of interest for organic strawberry production to be implemented in integrated control strategies for powdery mildew with potential to replace the use of pesticides and enhance product quality.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100313"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144587680","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Vaporized hydrogen peroxide sterilization of powered air-purifying respirators: Efficacy and safety assessment","authors":"Yeerzati Tuluhongtayi ,&nbsp;Wenjun He ,&nbsp;Kun Cai ,&nbsp;Weifang Han ,&nbsp;Sheng Zhang ,&nbsp;Jun Liu ,&nbsp;Wenwen Lei ,&nbsp;Zongxing Zhang ,&nbsp;Jiancheng Qi ,&nbsp;Guizhen Wu","doi":"10.1016/j.crbiot.2025.100312","DOIUrl":"10.1016/j.crbiot.2025.100312","url":null,"abstract":"<div><div>The COVID-19 pandemic increased demand for personal protective equipment, necessitating exploration of reprocessing methods. This study evaluates the efficacy and safety of vaporized hydrogen peroxide (VHP) sterilization for potential reuse of three powered air purifying respirator (PAPR) brands (3 M, Kebiao, Honeywell). <em>Geobacillus stearothermophilus</em> spores (ATCC 7953) served as biological indicators, while performance parameters (filtration, airflow, battery) and material integrity were monitored throughout 60 cycles, with H₂O₂ residual concentrations measured after 30 cycles. VHP achieved 100 % sterilization efficacy under both unloaded (48/48 biological indicators) and fully-loaded (132/132 biological indicators) conditions, demonstrating complete microbial inactivation across all sampling sites. However, brand-specific sterilization varied across operational conditions. 3 M units achieved complete sterilization in respiratory conduits when powered off but showed reduced filter efficacy. Kebiao units demonstrated complete sterilization when powered off but reduced efficacy during operation at internal filtration sites. Honeywell units with activated carbon showed poor VHP compatibility, while non-activated carbon units performed better when powered off. After 60 cycles, all PAPRs maintained filtration efficiency &gt; 99.97 % and airflow rates &gt; 120 L/min, with functional battery performance. H₂O₂ residual concentrations significantly exceeded safety thresholds (&lt;1 ppm): 3.2 (2.0,4.6) ppm for 3 M, 6.1 (4.3,7.4) ppm for Kebiao, and 6.5 (4.1,8.2) ppm for Honeywell units, with 3 M showing significantly lower residuals (P &lt; 0.05). This study provides critical evidence supporting VHP as a promising approach for PAPR sterilization. However, brand-specific protocols and extended ventilation procedures are required to ensure effective sterilization and reduce H₂O₂ residues to safe level.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100312"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144572717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeting the UCP1-dependent thermogenesis pathway with CRISPR/Cas9: a new approach to obesity management","authors":"Esmail Karami ,&nbsp;Fatemeh Rostamkhani ,&nbsp;Maasoume Abdollahi ,&nbsp;Mohamadreza Ahmadifard","doi":"10.1016/j.crbiot.2025.100295","DOIUrl":"10.1016/j.crbiot.2025.100295","url":null,"abstract":"<div><div>Obesity is a complex, multifactorial disease characterized by excessive body fat accumulation, which negatively impacts health. Its increasing prevalence has led to a global epidemic, emphasizing the urgent need for innovative and effective treatment strategies. This study aims to explore the potential of CRISPR/Cas9-mediated gene editing to enhance UCP1-dependent thermogenesis, offering a novel approach to obesity management. Uncoupling protein 1 (UCP1), primarily located in the inner mitochondrial membrane of brown adipose tissue (BAT), plays a crucial role in thermogenesis and energy expenditure. By converting stored energy into heat, UCP1 activation enhances calorie burning, helping to regulate body temperature and mitigate obesity-related health risks. Recent advancements in genome editing technologies, particularly CRISPR/Cas9, provide a precise method to modify genes involved in UCP1 expression and activity. This approach holds significant promise for sustainable obesity management by enhancing metabolic efficiency and energy expenditure. This study examines the feasibility of using CRISPR/Cas9 to target the UCP1-dependent thermogenesis pathway for obesity treatment. It explores the mechanisms of CRISPR/Cas9, the role of UCP1 in energy regulation, and potential strategies to enhance thermogenic activity. Our findings highlight the promise of CRISPR-based interventions in metabolic regulation. However, further research is necessary to optimize safety, efficacy, and regulatory considerations before translating these findings into clinical applications.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"9 ","pages":"Article 100295"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143916719","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}