Journal of Genetic Engineering and Biotechnology最新文献

{"title":"Comparative evaluation of E. coli expression systems for soluble hScFv-IL-17A with an unpaired CDR-L3 cysteine","authors":"Weeraya Thongkum ,&nbsp;Kanokporn Sornsuwan ,&nbsp;Thanathat Pamonsupornwichit ,&nbsp;Anuwat Weechan ,&nbsp;Natsima Viriyaadhammaa ,&nbsp;Kanchanok Kodchakorn ,&nbsp;Chatchai Tawapiwatana","doi":"10.1016/j.jgeb.2025.100613","DOIUrl":"10.1016/j.jgeb.2025.100613","url":null,"abstract":"<div><div>This study systematically evaluated <em>Escherichia coli</em> (<em>E. coli</em>) expression systems for the production of a humanized single-chain variable fragment targeting interleukin-17A (hScFv-IL-17A). A wild-type hScFv (hScFv-IL-17A-WT), derived from Secukinumab, was compared with a C97S mutant (hScFv-IL-17A-C97S) in which the unpaired cysteine residue in the light-chain complementarity-determining region 3 (CDR3) was substituted with serine. The primary objectives were to identify an optimal expression system for soluble protein production and to assess the functional consequences of the C97S mutation. Protein solubility, yield, and antigen-binding properties were evaluated across expression on three <em>E. coli</em> strains: Origami B (DE3), SHuffle, and BL21 (DE3) incorporating the DisCoTune system. Analyses were performed using SDS-PAGE, Western blotting, enzyme-linked immunosorbent assay (ELISA), and biolayer interferometry (BLI). Among the systems tested, BL21 (DE3) with DisCoTune yielded the highest levels of soluble expression for both variants. Although the C97S mutation enhanced soluble protein yield in this system, it significantly impaired functional activity. The hScFv-IL-17A-WT exhibited a strong binding affinity (K_D = 3.64 × 10⁻⁸ M), whereas the hScFv-IL-17A-C97S demonstrated an approximately 10-fold reduction in affinity. Competitive ELISA further confirmed that both variants recognized the same epitope as a neutralizing monoclonal antibody. Structural modeling predicted that the C97S substitution imposed a moderate stability penalty without inducing major conformational alterations, consistent with the observed reduction in binding affinity. Collectively, these findings indicate that while the C97S mutation improves soluble expression, it compromises biological activity. The hScFv-IL-17A-WT produced in BL21 (DE3) using the DisCoTune system represents the most promising candidate for subsequent therapeutic development.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100613"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145473965","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":"Detection method for CRISPR-Cas9 edited tomato with single nucleotide deletion using multiplex probe-based real-time PCR and confirmation of non-transgenicity: A case study","authors":"Monika Singh,&nbsp;Shiwani,&nbsp;Raghavendra Aminedi,&nbsp;Kushaldeep Kaur","doi":"10.1016/j.jgeb.2025.100609","DOIUrl":"10.1016/j.jgeb.2025.100609","url":null,"abstract":"<div><div>The adoption of gene-edited (GE) food crops has increased due to applicability of technology without drastically affecting the genetic information. However, the discussion on regulatory regime pertaining to GE plants throughout the globe is overwhelming. In the countries where GE products are regulated, or for the verification of gene-editing as claimed by the developers, detection of GE plants (referred to as GE detection here) plays an imperative role. Precisely detecting small modifications in single or few nucleotide(s) is a challenge. An efficient detection method was developed for GE tomato line with a single base pair edit as deletion in <em>Solanum lycopersicum pectate lyase</em> (<em>SlPL</em>) gene for better shelf life. A stepwise strategy is reported herein, comprising of screening at early phase targeting Cas9 protein gene using rapid loop-mediated isothermal amplification (LAMP) and conventional polymerase chain reaction (PCR) assays, followed by the verification of single point mutation using real-time PCR. Multiplex TaqMan® real-time PCR using fluorescent labelled dual probes simultaneously targeting edited and unedited sequences, was used for verification of single point deletion. This method was based on the negative selection where the presence of a mutation was determined by absence of signal in comparison to the wild-type. The developed multiplex real-time PCR method was found sensitive enough to detect 0.1% of the targeted lines. In view of the regulation of GM tomato in several countries, the approach for confirming non-transgenic nature of edited line was also presented using real-time PCR targeting common screening elements present in globally approved GM tomato events. This approach could be utilized as a tool globally either to support the regulatory authorities for food traceability or to verify the editing as well as to track suspected transgenic element in SDN-1 and SDN-2 types.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100609"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145424507","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":"Prognostic and functional role of PPAR-alpha and SNP receptor (Leu162Val) in acute coronary syndrome: a potential novel target","authors":"Abdullah Abdulsattar Raeef ,&nbsp;Hassan H. Al-Saeed ,&nbsp;Sami Mekhlif Mishlish","doi":"10.1016/j.jgeb.2025.100555","DOIUrl":"10.1016/j.jgeb.2025.100555","url":null,"abstract":"<div><h3>Background and objective</h3><div>PPAR-alpha plays a key role in acute coronary syndrome (ACS). Its activation influences lipid metabolism and inflammation, impacting cardiovascular health and potentially mitigating risks related to heart conditions. This study aims to find the relationship between PPAR-alpha concentration and genetic polymorphism in ACS patients.</div></div><div><h3>Methods and materials</h3><div>The study enrolled 90 ACS patients and 90 controls aged 30–70. Blood samples were analyzed for biomarkers (troponin, CK-MB, hs-CRP, PPARα) and rs1800206 SNP via ELISA and HRM-PCR. Data were statistically analyzed using SPSS v21, ANOVA, and ROC tests.</div></div><div><h3>Results</h3><div>The results showed an increase in the levels of troponin, CK-MB, and hs-CRP for the patient groups compared to the healthy groups with significant differences between the groups (p-value &lt;0.05). The level of (PPAR-alpha) was low in patients (1472.2 ± 74.5) and high in the healthy group (2051.8 ± 149.9) with significant differences between study groups. ROC analysis identified CK-MB, and hs-CRP as optimal biomarkers for ACS diagnosis; PPARα showed good predictive value. The study also found that the dominant genotype (CC) represented a small percentage of the patient samples while the homozygotes (CG) and the recessive (GG) represented the highest percentage, unlike the healthy group. Also, Allele (C) has a low percentage in the patients’ group while allele (G) has a high percentage with considering the G allele as a risk factor. In addition, the results showed an abnormal distribution of Hardy-Weinberg equilibrium indicating the presence of factors extraneous to the community that changed the genetic pattern. The results also showed that the dominant genetic model is the best expressive for the samples of the Iraqi community infected with ACS.</div></div><div><h3>Conclusions</h3><div>The present study demonstrated a relationship between the concentration of PPAR-alpha and the genetic polymorphism of Leu162Val (rs1800206) SNP and acute coronary syndrome with a decrease in the concentration of PPAR-alpha and the predominance of the recessive gene and the recessive allele in patients compared to healthy individuals.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100555"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144879271","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":"Transcriptome profiling by RNA sequencing reveals novel targets of Gemini nano curcumin on p53-mutant HT-29 colorectal cancer cells","authors":"Hewa Jalal Azeez ,&nbsp;Raheleh Karimzadeh ,&nbsp;Esmaeil Babaei","doi":"10.1016/j.jgeb.2025.100548","DOIUrl":"10.1016/j.jgeb.2025.100548","url":null,"abstract":"<div><h3>Introduction</h3><div>Colorectal cancer is considered as an aggressive tumor with high mortality in the world. It has been shown that Gemini Nano-Curcumin (Gemini-Cur) affects viability of colorectal cancer cells. Nevertheless, the cellular and molecular mechanisms underlying its toxicity are debatable. Here, we planned to untangle the potential novel targets for nanocurcumin on p53-mutant HT-29 cancer cells by employing RNA sequencing.</div></div><div><h3>Methods</h3><div>After cultivation, HT-29 cell were incubated with appropriate doses of Gemini-Cur for 24 h. Total RNA was extracted, cDNA library was constructed and sequenced. The DESeq2 tool were employed to normalize reads and detect differentially expressed genes (DEGs). The enrichR tool was employed to do Gene Ontology (GO) &amp; identify biological processes (BP), cellular components (CC) and molecular functions (MF) that are impacted in the condition. The PPI network was constructed of 1200 DEGs using STRING, visualized by Cytoacape and analyzed with MCODE.</div></div><div><h3>Results</h3><div>After DEGs screening, 1309 genes between untreated and treated cancer cells were obtained including 479 upregulated with P-value &lt; 0.05 &amp; log2 FC &gt; 1) as well as 63 downregulated genes with P-value &lt; 0.05 and log2 FC &lt; −1). Then, DEGs were assigned to 207 GO terms including numerous cellular pathways such as endoplasmic reticulum (ER)-related processes. Finally, 542 up and downregulated genes were mapped to 67 reactome pathways. The pathway analysis illustrated that Gemini-Cur modulates numerous pathways including ER stress response- and transporter-related genes. Using MCODE, Three modules were significantly identified with scores ≥ 1 and nodes ≥ 1.</div></div><div><h3>Conclusion</h3><div>Our data reveal that nanocurcumin might affects HT-29 colorectal cancer cells through modulating different pathways such as endoplasmic reticulum response, and transporter-related genes. More studies necessitate to unravel the molecular mechanisms and proteins involved in these pathways that could be considered as novel therapeutic targets in colorectal cancer.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100548"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144867229","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":"Isolation and characterization of chitinolytic fast-growing bacteria from marine environment in Vietnam","authors":"Thi Van Anh Tran ,&nbsp;Thuy Hang Dam ,&nbsp;Lan Huong Nguyen ,&nbsp;Tuan Le ,&nbsp;Thi Kieu Oanh Vu ,&nbsp;Thomas Walther ,&nbsp;Ha Thanh Le","doi":"10.1016/j.jgeb.2025.100587","DOIUrl":"10.1016/j.jgeb.2025.100587","url":null,"abstract":"<div><div>Chitin is found in tremendous quantities in nature, especially in the marine environment. This polymer has a stable structure that requires a relatively long time to degrade. As a result, fast-growing microorganisms capable of degrading chitin might be a potential approach in responding to the hydrolysis of this abundant material. This study focused on the isolation of chitinolytic fast-growing bacteria from marine environment in Vietnam, determined their biochemical characteristics, and generally evaluated the relationship between the presence of some important chitinase genes (mostly extracellular) and chitin degradation capacity of the isolates. Crucial chitinase genes were detected via PCR using degenerate primers, while chitin hydrolysis ability was determined via enzymatic measurements, assessing strains’ growth on chitin and chitin hydrolysate composition. The results showed that fourteen chitinolytic fast-growing bacteria isolated from seawater in Vietnam belonged to the family <em>Vibrionaceae</em> and the majority of them possessed all five tested chitinase genes including <em>VCA0027</em> (chitinase), <em>VC1952</em> (endochitinase), <em>VC0769</em> (endochitinase), <em>VCA0700</em> (chitodextrinase), <em>VC2217</em> (hexosaminidase). <em>V. natriegens</em> isolates N5.3, N7.2, and 10.3, which lack VC1952 and VC0769, exhibited significantly reduced enzymatic activities, weak chitin hydrolysis, and limited growth on chitin substrates, highlighting the crucial role of these genes for efficient chitin degradation.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100587"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145362168","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":"Hub genes and common pathogenic mechanisms between COPD and H. pylori infection","authors":"Keying Li ,&nbsp;Tuliang Liang ,&nbsp;Minfang Li ,&nbsp;Xuxin Sun ,&nbsp;Wei Xie ,&nbsp;Sheng Chen","doi":"10.1016/j.jgeb.2025.100622","DOIUrl":"10.1016/j.jgeb.2025.100622","url":null,"abstract":"<div><h3>Background</h3><div>Chronic obstructive pulmonary disease (COPD) and<!--> <em>Helicobacter pylori</em> (<em>H. pylori</em>) infection may share common pathogenic mechanisms, yet their molecular and cellular interactions remain poorly defined.</div></div><div><h3>Methods</h3><div>Public datasets for COPD and <em>H. pylori</em> <!-->infection were retrieved from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified, followed by weighted gene co-expression network analysis (WGCNA) to define disease-associated modules. Overlapping genes underwent functional enrichment via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Support vector machine-recursive feature elimination (SVM-RFE) was analyzed<!--> <!-->to identify hub genes. Gene set enrichment analysis (GSEA), receiver operating characteristic (ROC) curve validation, and immune infiltration analysis using CIBERSORT were performed. Single-cell RNA sequencing (scRNA-seq) and cell–cell communication analysis were further applied to characterize hub gene expression and intercellular interactions.</div></div><div><h3>Results</h3><div>Integrative analysis identified 100 co-expressed genes, which were associated with cytokine-receptor interaction, TNF signaling, and chemokine-mediated signaling.<!--> <!-->SVM-RFE highlighted<!--> <!-->CCR1<!--> <!-->and<!--> <!-->CCL19<!--> <!-->as shared genes, validated in external cohorts, with GSEA indicating involvement in immune and inflammatory signaling.<!--> <!-->ROC curves confirmed robust diagnostic performance.<!--> <!-->Immune infiltration analysis showed macrophages M0 were consistently elevated in COPD and <em>H. pylori</em> infection; CCR1 and CCL19 are associated with multiple immune cell types. Single-cell analysis revealed hub genes are primarily expressed in macrophages and fibroblasts, potentially modulating pathogenesis via interactions between immune and signaling pathways.</div></div><div><h3>Conclusion</h3><div>CCR1 and CCL19 are shared genes linking COPD and <em>H. pylori</em> infetion, which regulate immune homeostasis and inflammatory signaling<!--> <!-->via influencing macrophages<!--> <!-->and fibroblasts. These findings offered new ideas for molecular mechanisms and integrated therapeutic interventions<!--> <!-->in COPD and <em>H. pylori</em> infetion.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100622"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145623559","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":"Potential antibacterial, antioxidant and anticancer activities of Nigerian mountain honey in relation to its bacterial flora","authors":"Sara H. Mansour ,&nbsp;Asmaa I. El-shazly ,&nbsp;Amira A. Gamal ,&nbsp;Elsayed A. Elsayed ,&nbsp;Mohammad A. Wadaan ,&nbsp;Mona A. Esawy","doi":"10.1016/j.jgeb.2025.100559","DOIUrl":"10.1016/j.jgeb.2025.100559","url":null,"abstract":"<div><div>Numerous studies have investigated the biological activities of honey, emphasizing its antimicrobial, antioxidant, and therapeutic properties. However, a comprehensive examination of the bacteria isolated from honey has been largely overlooked. This study aimed to bridge that gap by exploring the relationship between the bioactivities of honey and the bacterial spores it contains. Furthermore, there is no comparison between the polysaccharides in honey such as levan and its inherent bacteria. Accordingly, various enzymes, including levansucrase, invertase, glucose oxidase, amylase, and catalase, were assessed in Nigerian mountain honey and their inherent seven bacterial isolates. Honey and its isolates showed levansucrase production to various degrees where the highest activity was obtained with isolate NO.4 and the honey recorded (21U/ml). Additionally, honey and its isolates showed varied activities between (171–465 U/ml), (88–250 U/ml) for amylase and glucose oxidase respectively. The highest catalase activity was recorded by isolate No 4 based on the qualitative test. Interestingly, all isolates lacked protease activity. Honey and specific isolates inhibited various pathogenic bacteria, including <em>Salmonella enterica Typhi, Escherichia coli,</em> and <em>Bacillus cereus.</em> Also, fungi such as <em>Candida albicans.</em> In addition, honey and its isolates had high antioxidant activities ranging from 58 to 77%. The polysaccharides Lev1, Lev2, and honey have no cytotoxic effects on normal cells. They were identified as levans based on FTIR and NMR. Honey and lev1, lev2 showed cytotoxicity against HCT116 cell lines (61.5, 59.5, and 53.70%), respectively. Interestingly, the study found a strong link between the biological activities of the bacteria and the honey itself.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100559"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144896453","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":"The effects of gut microbiota-derived short-chain fatty acids (SCFAs) on human colon cancer cells: An in vitro study","authors":"Shiva Darabi ,&nbsp;Fatemeh Keshavarzi ,&nbsp;Parviz Ashtari ,&nbsp;Farahnaz Motamedi Sedeh ,&nbsp;Behrouz Alirezapour","doi":"10.1016/j.jgeb.2025.100610","DOIUrl":"10.1016/j.jgeb.2025.100610","url":null,"abstract":"<div><div>Colorectal cancer (CRC) has always been a major concern for researchers due to its deadly nature. One of the microbial metabolites found in the gut is short-chain fatty acids (SCFAs), which have the ability to regulate the immune system and exert protective effects against CRC. In this study, we investigated the effects of SCFAs on the expression of the <em>CYP1A1</em> gene in Caco-2 cells at varying concentrations. The Caco-2 cell line and HDF control cell line were treated with three concentrations (7.5, 15, and 30 mM) of NaA(sodium acetate), NaB(sodium butyrate), and NaP(sodium propionate) for three different time periods (24, 48, and 72 h). Additionally, two concentrations (2.5 and 10 μM) of CH223191 (an AhR gene antagonist) alone and a concentration of 10 μM of CH223191 in combination with the mentioned concentrations of NaA, NaB and NaP were applied to the cells for 48 h. The gene expression of <em>CYP1A1</em> was examined using Real-time PCR. The expression of the <em>CYP1A1</em> gene increased with concentrations of NaA and NaB at all time points, and with 30 mM of NaP at 24 h (p &lt; 0.05). However, increasing the concentration of NaB and NaP led to decreased expression of the <em>CYP1A1</em> gene. These findings suggest the potential use of SCFAs as epigenetic drugs for the prevention and treatment of CRC.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100610"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145424546","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":"In vitro antiproliferative effects of green synthesized silver nanoparticles from Brassica carinata microgreens on DU-145 prostate cancer cells and In vivo safety assessment","authors":"Dogfounianalo Somda ,&nbsp;Joel L. Bargul ,&nbsp;Sabina Wangui Wachira ,&nbsp;John M. Wesonga","doi":"10.1016/j.jgeb.2025.100552","DOIUrl":"10.1016/j.jgeb.2025.100552","url":null,"abstract":"<div><h3>Background</h3><div>Prostate cancer ranks among the most lethal malignancies affecting men globally. Conventional therapeutic approaches for this disease encounter limitations due to adverse side effects and the emergence of drug resistance, making its management challenging. Green-synthesized silver nanoparticles (AgNPs) have emerged as an attractive alternative to address the shortcomings of existing anticancer treatments.</div></div><div><h3>Objective</h3><div>This study was undertaken to evaluate the anticancer potential and safety profile of <em>BCM-</em>AgNPs, focusing on DU-145 inhibition and sub-acute toxicity in Wistar rats.</div></div><div><h3>Methods</h3><div>Previously synthesized <em>Brassica carinata</em> microgreen silver nanoparticles (<em>BCM</em>-AgNPs) alongside <em>Brassica carinata</em> microgreen crude extract (BCME) were used. The antiproliferative activity was investigated through the MTT assay on DU-145 prostate cancer cells and normal Vero E6 cells to determine the IC₅₀ and CC₅₀. Subsequently, a series of bioassays, including cell migration assays, clonogenic assays, cell cycle assays, and DNA fragmentation assays, were conducted. Furthermore, RT-qPCR was performed to delineate the molecular mechanism of <em>BCM</em>-AgNPs on DU-145 cells. Finally, a subacute toxicity study was carried out to assess the safety of <em>BCM</em>-AgNPs.</div></div><div><h3>Results</h3><div>Both <em>BCM</em>-AgNPs and BCME exhibited potent antiproliferative activity against DU-145 cells with an IC₅₀ of 33.47 μg/mL and 46.25 μg/mL, respectively, while sparing normal Vero E6 cells with selectivity indices of 3.50 for <em>BCM</em>-AgNPs and 4.62 for BCME. <em>BCM-AgNPs</em> demonstrated anti-migratory effects and hindered colony formation in DU-145 cells. They further induced S-phase cell cycle arrest through DNA fragmentation. RT-qPCR analysis revealed the upregulation of pro-apoptotic genes and the downregulation of genes involved in cell survival and proliferation pathways. <em>BCM</em>-AgNPs showed no toxicity in Wistar rats during 28-day oral administration.</div></div><div><h3>Conclusion</h3><div>This study demonstrates for the first time the selective and prominent antiproliferative effects of <em>BCM</em>-AgNPs against DU-145, as well as their safety and biocompatibility, underscoring their potential as a promising candidate for anticancer therapy.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100552"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145105183","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":"Optimization of fermentation conditions for enhanced L-arginase production by Alcaligenes aquatilis BC2 using response surface methodology","authors":"Birhan Getie Assega ,&nbsp;Kefyalew Ayalew Getahun ,&nbsp;Tamene Milkessa Jiru ,&nbsp;Tsehayneh Geremew Yohannes ,&nbsp;Mulugeta Aemero ,&nbsp;Berhanu Andualem","doi":"10.1016/j.jgeb.2025.100591","DOIUrl":"10.1016/j.jgeb.2025.100591","url":null,"abstract":"<div><div>L-arginase-based enzyme therapy, which depletes L-arginine by converting it to L-ornithine and urea, selectively inhibits the growth of L-arginine-dependent cancer cells with low toxicity. This approach shows promise as a novel cancer treatment. This research used Response Surface Methodology (RSM) to enhance L-arginase production by <em>Alcaligenes aquatilis</em> BC2, which was isolated from an Ethiopian soda lake. The Plackett-Burman Design was used to screen eight factors that influence L-arginase production and identified arginine concentration, peptone concentration, and incubation temperature as the most significant variables. The central composite design analysis demonstrated that the optimized conditions of 1.75 % L-arginine concentration, 3 % peptone concentration, and an incubation temperature of 37.5 °C enhance L-arginase production from a baseline of 92.45 U/mL to an optimized yield of 288.79 U/mL. This represents a 3.1-fold increase under the optimized conditions.</div><div>The model was developed based on 20 experimental runs, demonstrating excellent fit with R2 = 0.9974 and a significant F-value of 420.28 (p &lt; 0.0001). Additionally, the lack-of-fit test was conducted and found to be non-significant (F-value = 4.18, p = 0.0714), further supporting the model’s predictive strength. This investigation showed that applying statistical design to optimize fermentation conditions leads to increased production of L-arginase, thereby advancing enzyme-based therapeutic practices and highlighting statistical optimization as essential for bioprocess development.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100591"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145332238","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}