Applied Biochemistry and Biotechnology最新文献

{"title":"Characterization of Novel Pullulanase Type I from Newly Isolated Bacillus cereus Strain NP9: Potential Additive for Laundry Detergent Formulations","authors":"Nihan Arabaci","doi":"10.1007/s12010-025-05286-1","DOIUrl":"10.1007/s12010-025-05286-1","url":null,"abstract":"<div><p>This study aimed to produce a pullulanase that can be utilized as an additive in detergent formulations. A newly isolated <i>Bacillus cereus</i> strain NP9 exhibited the highest pullulanase activity and was selected for production. The optimum conditions for crude NP9 pullulanase were a pH of 7.0 and a temperature of 40 °C. It maintained stability at high rates within the pH range of 5.0 to 11.0 and temperatures between 25 and 65 °C. The molecular weight of the enzyme was determined to be approximately 170 kDa via native-PAGE. Thin-layer chromatography and high-performance liquid chromatography analyses indicated that NP9 pullulanase converted pullulan and starch substrates into maltotriose units (pullulanase type I). The enzyme exhibited moderate activity with certain metal ions and was not Ca²⁺-dependent. The inhibition of the enzyme by EDTA, EGTA, and 1,10-phenanthroline indicated it is a metalloenzyme. The enzyme moderately retained activity when exposed to non-ionic detergents such as Triton X-100, Tween 20, and Tween 80. It demonstrated high compatibility (90%) with the commercial detergent “Peros.” Wash performance analyses showed that the NP9 pullulanase and commercial detergent mixture removed starchy stains more effectively than washing with commercial detergent alone. In conclusion, NP9 pullulanase exhibited favorable properties, making it a potential candidate for the laundry detergent industry.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5470 - 5493"},"PeriodicalIF":3.3,"publicationDate":"2025-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12334528/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144324092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Anti-Melanogenesis Activity of Peptides from Shark (Mustelus griseus) Skin on B16F10 Melanocytes and In vivo Zebrafish Models","authors":"Yonghua Hu,&nbsp;Weichao Su,&nbsp;Baoxuan Wang,&nbsp;Xiaoyu Yang,&nbsp;Lina Ke,&nbsp;Yan Shi,&nbsp;Yuleyue Wang,&nbsp;Qin Wang","doi":"10.1007/s12010-025-05296-z","DOIUrl":"10.1007/s12010-025-05296-z","url":null,"abstract":"<div><p>Shark skin peptides, a kind of substance with a variety of biological activities, were shown to inhibit the activity of mushroom tyrosinase. This study was performed to clarify their inhibitory effects on melanin synthesis in B16F10 melanocytes and in vivo zebrafish models. In order to elucidate the inhibitory effects of shark skin peptides on B16F10 melanocytes, cell viability, morphologic changes, intracellular tyrosinase, and the melanogenesis-related proteins were determined. The results showed that the peptides were not cytotoxic to B16F10, and the morphology of the cells did not change significantly at the detected concentration. The intracellular melanin content and tyrosinase activity of B16F10 cells decreased in a concentration-dependent manner after the peptide treatment. The suppressive activity of the peptides on melanin synthesis is linked to the down-regulation of the important proteins involved in melanogenesis, such as tyrosinase, tyrosinase-related protein-1 (TRP1), and microphthalmia-associated transcription factor (MITF). The effects of shark skin peptides on melanin synthesis in zebrafish embryos showed that the peptides inhibit the production of melanin in embryos, and the melanin-related proteins were also detected as down-regulated. The results from ROS analysis suggested that the peptides are also capable of eliminating reactive oxygen species within the cells. In conclusion, the shark skin peptides could inhibit melanin synthesis, indicating that peptides may become additives for cosmetics potentially.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5396 - 5410"},"PeriodicalIF":3.3,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144315685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of Secondary Metabolites in Reducing the Negative Impact of Pest-Induced Stress in Eggplant (Solanum melongena L.)","authors":"Pratik Talukder,&nbsp;Srishti Chakraborty,&nbsp;Mainak Sarkar,&nbsp;Anirban Das,&nbsp;Rajarshi Ray","doi":"10.1007/s12010-025-05287-0","DOIUrl":"10.1007/s12010-025-05287-0","url":null,"abstract":"<div><p>Brinjal, also known as eggplant, is a significant crop in the Solanaceae family, with a high economic value and widespread cultivation in Indian states. Its primary metabolites include carbohydrates, proteins, lipids, organic acids, and polyphenolic compounds. Secondary metabolites contribute to the fruit’s flavor, color, and potential health benefits, such as chlorogenic acid, anthocyanins, flavonoids, alkaloids, and other phytochemicals. The phenylpropanoid pathway is a metabolic pathway in plants that produces a broad spectrum of secondary metabolites, including phenolic compounds, lignins, flavonoids, and other compounds. In brinjal fruit, the phenylpropanoid pathway is important for the synthesis of secondary metabolites, such as phenolic compounds and lignins, which contribute to the fruit’s color, flavor, and potential health benefits. Biological pests, such as shoot borer and fruit borer (<i>Leucinodes orbonalis</i>), can cause significant damage to crops, resulting in yield losses and economic consequences for farmers. The shoot borer and fruit borer follow a typical life cycle, starting with adult caterpillars laying eggs on the surface of young leaves and shoots of eggplant. After hatching, larvae burrow into plant tissue, creating tunnels in the buds and fruit, causing fruit wilting, deformity, and stunting. Ingestion weakens plants, making them more susceptible to secondary bacterial and fungal infections. Young shoots and fruit borer management can be difficult due to their hidden feeding habits. Integrated pest management (IPM) strategies are commonly used to effectively control this pest, which includes a combination of farming practices, biological control, and the rational use of chemical pesticides. This scientific investigation sincerely aims to attain a deep knowledge of the myriad ways pests affect and how this information can guide recovery and mitigation strategies. The design of these experiments acts as a potential tool for analyzing compound ongoing dynamics, unraveling the complexity of plant–pesticide interactions.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5411 - 5426"},"PeriodicalIF":3.3,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144315710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing Biohydrogen Yields: A Comparative Study of Individual and Combined Biomass Pretreatment Techniques","authors":"Chelladurai Mumtha,&nbsp;Pambayan Ulagan Mahalingam","doi":"10.1007/s12010-025-05298-x","DOIUrl":"10.1007/s12010-025-05298-x","url":null,"abstract":"<div><p>Biohydrogen production from various biomass sources using combined pretreatment methods is an emerging and cost-effective alternative energy technology. To enhance hydrogen production, a batch test was conducted involving heat treatment, ultrasonication, and acid hydrolysis of dairy whey (DW) and sugarcane bagasse (SCB). The heat-treated DW achieved a maximum cumulative hydrogen production of 153.4 ± 2.0 mL H₂/L, which is 20% higher than the untreated biomass. The physicochemical changes in both DW and SCB were analyzed using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive X-ray analysis (EDAX). The tools used to study the effects of pretreatments on SCB were X-ray diffraction (change in crystalline and amorphous regions) and SEM images. FTIR spectra showed the removal of hemicellulose and lignin content during pretreatments. The presence of cellulose, hemicellulose, and lignin structures in the sample of sugarcane bagasse and cellulosic fractions were indicated by the characteristic absorption bands (cm⁻¹) of groups shown by the FTIR spectrum for hemicellulose and lignin. SEM images showed extensive degradation of the buddle of fibers of some cellulosic fraction. FTIR spectra indicate that carbohydrates, proteins, and lipids were present in dairy whey. The EDX results indicated that untreated SCB contains 30% carbon, 13.71% oxygen, and 0.50% nitrogen. SCB underwent acid hydrolysis for 90 min at 121 °C using a 2 M concentration of H₂SO₄. The highest cumulative hydrogen production of 189.6 ± 4.3 mL H₂/L was achieved at 37 °C under co-culture conditions. Based on the findings of this study, it may be possible to produce biohydrogen from biomass in a manner that is both efficient and sustainable in the future.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5427 - 5451"},"PeriodicalIF":3.3,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144315709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization and Function of a Novel Extracellular Polysaccharide from a Green Alga Parachlorella sp. AMI5","authors":"Reo Yamane,&nbsp;Reiya Ishida,&nbsp;Yuuya Uejima,&nbsp;Akari Takagi,&nbsp;Akihiro Nakamura,&nbsp;Munehiko Asayama","doi":"10.1007/s12010-025-05276-3","DOIUrl":"10.1007/s12010-025-05276-3","url":null,"abstract":"<div><p>The green alga <i>Parachlorella</i> sp. AMI5 is a newly identified strain that produces a substantial amount of the unique extracellular polysaccharide amiEPS. This study aimed to investigate the purification, characteristics, and function of amiEPS-N from the AMI5 strain cultivated in a nitrogen-deficient BG11 (BG11-N) culture. We successfully obtained a high yield of 1.08 g/L of purified amiEPS-N with low protein content. Characteristic analyses revealed that amiEPS-N was composed of a flexible molecular form with 0.51 ± 0.00 log (nm)/log (g/mol) of a conformation plot slope in the water solution, and the weight average molecular mass was 1.73 × 10⁶ g/mol in maximum. The monosaccharide composition of amiEPS-N was rhamnose:xylose:glucuronic acid as 55.1:34.8:10.1 (mol%), indicating that it is an acidic rhamnan. The aqueous amiEPS-N exhibited anti-angiotensin-converting enzyme and tyrosinase-enhancing activities, which may contribute to the anti-hypertensive effects and inhibition of gray hair, indicating its novel and unique properties. The biochemical characteristics of the novel amiEPS and the contribution of this production technology to the social implementation for biorefineries are discussed.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5378 - 5395"},"PeriodicalIF":3.3,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144265004","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcription Factor TCF12 Activates the Transcription Level of SLC38A1 to Promote the Development of Hepatocellular Carcinoma","authors":"Xiaochun Li,&nbsp;Min Liang,&nbsp;Zizhong Xu,&nbsp;Zhili Wei,&nbsp;Yu Xu","doi":"10.1007/s12010-025-05263-8","DOIUrl":"10.1007/s12010-025-05263-8","url":null,"abstract":"<div><p>Hepatocellular carcinoma (HCC) is a primary liver cancer with a high mortality rate. The pathogenic mechanism of HCC is complex. In this study, we aimed to explore the functions of Solute Carrier Family 38 Member 1 (SLC38A1) and transcription factor 12 (TCF12) in HCC malignancy. Immunohistochemistry (IHC) assay was performed to estimate the expression of SLC38A1 in HCC. qRT-PCR assay and western blot assay were conducted to determine the expression of SLC38A1, TCF12, epithelial-mesenchymal transition (EMT)-related markers and ferroptosis-related markers. Colony formation assay and EdU assay were utilized to evaluate cell proliferation ability. Transwell assay was employed for cell migration and invasion. Reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and iron levels were examined with related kits. ChIP assay and dual-luciferase reporter assay were performed to verify the relationship between TCF12 and SLC38A1. The in vivo experiment was conducted to assess the function of TCF12 in vivo. SLC38A1 was upregulated in HCC tissues and cells. SLC38A1 knockdown suppressed HCC cell proliferation, migration, invasion, and EMT and inhibited ferroptosis in vitro. The transcription factor TCF12 could activate the transcription level of SLC38A1. TCF12 overexpression ameliorated the effects of SLC38A1 knockdown on HCC cell malignant behaviors. Moreover, TCF12 silencing inhibited tumorigenesis in vivo. TCF12 targeted SLC38A1 to promote HCC cell proliferation, migration and invasion and restrained ferroptosis, providing a novel sight for HCC pathogenesis.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5362 - 5377"},"PeriodicalIF":3.3,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144232877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stability of Burkholderia cepacia Lipase Immobilized on Styrene–Divinylbenzene Activated with Glutaraldehyde, Triton X-100, and Polyethylene Glycol for the Green Synthesis of Hexyl Acetate","authors":"Wellington M. Correa,&nbsp;Wislei R. Osório,&nbsp;Ausdinir D. Bortolozo,&nbsp;Erik Poloni,&nbsp;Giovana S. Padilha","doi":"10.1007/s12010-025-05277-2","DOIUrl":"10.1007/s12010-025-05277-2","url":null,"abstract":"<div><p>This work focuses on improving the stability of <i>Burkholderia cepacia</i> lipase immobilized on styrene–divinylbenzene by using chemical additives and a cost-effective physical adsorption method. Ethanol pretreatment of the supports proved essential for maintaining enzyme activity. The optimal conditions for immobilization were achieved at a 1:1 support-to-enzyme ratio, pH 8, 200 rpm, and 60 °C. Combinations of the additives glutaraldehyde, polyethylene glycol 1500, and Triton X-100 were examined for activation treatment of supports before immobilization. Concentrations of 2.5% (w/v) of polyethylene glycol 1500 and 0.5% (v/v) of Triton X-100 were used to maximize biocatalyst activity. We show that the activated biocatalyst yielded up to 950% more hexyl acetate than non-activated control after 12 reaction cycles. Fourier transform infrared spectroscopy and scanning electron microscopy confirmed the effective immobilization of the <i>Burkholderia cepacia</i> lipase. This study introduces a scalable and sustainable method for creating robust biocatalysts aimed at producing value-added chemicals, thereby advancing green chemistry in the flavor industry.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5344 - 5361"},"PeriodicalIF":3.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12334490/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Relationship Between Activating Transcription Factor 3 and Forkhead Box Protein A2 in Spinal Cord Injury and the Underlying Mechanism","authors":"Xiong Dong,&nbsp;Huaizhi Gu,&nbsp;Guanhua Xu,&nbsp;Hongxiang Hong,&nbsp;Zhiming Cui","doi":"10.1007/s12010-025-05256-7","DOIUrl":"10.1007/s12010-025-05256-7","url":null,"abstract":"<div><p>Activating transcription factor 3 (ATF3) may function as a regulator of various diseases; however, its role in spinal cord injury (SCI) remains unknown. We designed a current work to evaluate the potentials of the ATF3/forkhead box protein A2 (FOXA2) axis in SCI. GSE45006 chip was analyzed, and a volcano plot and heatmap were drawn. Gene Ontology and KEGG analysis were performed for the differentially expressed genes. Animals with SCI were established. Quantitative reverse transcription polymerase chain reaction and western blotting were used to determine mRNA expression, and western blotting was used for detecting protein expression. The interaction between FOXA2 and the ATF3 promoter was evaluated using the UCSC database and confirmed using dual-luciferase and chromatin immunoprecipitation assays. Cellular behaviors were determined using CCK-8, EdU, and TUNEL assays. Levels of p-PI3K, PI3K, p-AKT, and AKT were examined by the WB method. We found that ATF3 expression was markedly increased in rats with SCI. Interestingly, ATF3 knockdown increased the proliferation and suppressed the apoptotic ability of PC12 cells. FOXA2 activates ATF3 transcription. Knockdown of FOXA2-mediated down-regulation of ATF3 increases growth and decreases PC12 cell death. ATF3 knockdown could increase the level of p-PI3K and p-AKT; FOXA2 shRNA could affect the expression of p-PI3K and p-AKT, which was partially abrogated by ATF3 OE. Forkhead box protein regulates the transcription of ATF3, thereby affecting cell growth and PC12 cell death.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5327 - 5343"},"PeriodicalIF":3.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214557","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrated Analysis of Proteome and Metabolome Reveals the Basis of Amino Acid Metabolism in Cigar Artificial Fermentation","authors":"Shujun Chen,&nbsp;Fuxiang Zhu,&nbsp;Shengkui Zhang,&nbsp;Shengxiao Wang,&nbsp;Yanyuan Shen,&nbsp;Mengmeng Zhang,&nbsp;Wenxiao Hu,&nbsp;Qingxiang He,&nbsp;Lei Qiu,&nbsp;Qidong Hao,&nbsp;Zhixing Li,&nbsp;Zhao Liu,&nbsp;Yvqing Ding,&nbsp;Meng Xu,&nbsp;Hongying Kan,&nbsp;Yanqi Hu,&nbsp;Xianyan Zhao","doi":"10.1007/s12010-025-05275-4","DOIUrl":"10.1007/s12010-025-05275-4","url":null,"abstract":"<div><p>Cigars are a type of tobacco product made entirely from dried tobacco, primarily consisting of the filler, binder, and wrapper. Fermentation is a key step in improving the quality of cigar tobacco leaves (CTLs). To investigate how fermentation affects quality, this study employed non-targeted metabolomics and data-independent acquisition (DIA) proteomics to examine the metabolic changes and protein expression levels in tobacco leaves. The results reveal that a total of 112 differential metabolites were identified through untargeted metabolomics, with 87 compounds demonstrating a decrease in relative abundance post-fermentation, including 20 amino acids and their derivatives. Utilizing DIA proteomics, 341 differentially expressed proteins were identified. Functional analysis of these proteins revealed variations in biological functions at different fermentation stages. A total of 21 driver proteins exhibited significant correlations with the metabolic regulation of eight amino acids. This study revealed that the transformation of amino acid metabolism significantly affects the quality of CTLs. It enhanced the understanding of amino acids among the differential metabolites before and after fermentation. This research provides a theoretical basis for the control of amino acids during the artificial fermentation process of CTLs, aiming to further improve their quality.</p><h3>Graphical Abstract</h3>\u0000<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5308 - 5326"},"PeriodicalIF":3.3,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12010-025-05275-4.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144155344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Alhagi maurorum: A Medicinal Treasure Trove Empowered by Copper Oxide Nanoparticles for Enhanced Secondary Metabolite Synthesis","authors":"Deepak Bamal,&nbsp;Anoop Singh,&nbsp;Nisha Swami,&nbsp;Anita Rani Sehrawat","doi":"10.1007/s12010-025-05284-3","DOIUrl":"10.1007/s12010-025-05284-3","url":null,"abstract":"<p>This study investigated the potential of green-synthesized copper oxide nanoparticles (CuO NPs) to enhance biomass production and therapeutic metabolite yields in <i>Alhagi maurorum,</i> a medicinal plant of significant pharmaceutical value. CuO NPs were biosynthesized using <i>A. maurorum</i> leaf extract as a reducing and capping agent, with characterization confirmed via UV-Vis spectroscopy, FTIR, XRD, SEM, TEM, and zeta potential analysis. Nanoparticles ranged from 7-30 nm in size. Callus induction and proliferation were established using Murashige and Skoog (MS) media supplemented with varying concentrations (0-12 mg/L) of CuO NPs combined with plant growth regulators. Maximum callus fresh weight (9.02 mg in cotyledon and 8.46 mg in hypocotyl) was achieved in MS media containing 3.0 mg/L BAP, 0.1 mg/L NAA, and 0.50 mg/L kinetin without CuO NPs. However, CuO NPs significantly enhanced metabolite production in a dose-dependent manner. Analysis of variance revealed statistically significant differences (<i>p</i>=0.001) across all biochemical parameters tested, with high <i>F</i>-values for peroxidase activity (7,755.74), total flavonoids (5,195.02), and total soluble sugar (5,702.18). At 8 mg/L CuO NPs, callus cultures exhibited elevated levels of total free amino acids (12.49±0.023 mg/g DW) and total soluble protein (35.617±0.033 mg/g DW), while control samples produced higher starch (35.547±0.23 mg/g DW) and total soluble sugar (121.56±0.091 mg/g DW) content. Significantly, CuO NP-treated cultures demonstrated enhanced secondary metabolite synthesis, with maximum total phenolic compounds (156.477±0.167 mg/g DW GAE) and flavonoids (58.307±0.179 mg/g QE) at 8 and 10 mg/L CuO NPs, respectively. Antioxidant enzyme analysis revealed that cotyledon-derived callus exhibited peak activities at specific CuO NP concentrations: superoxide dismutase (84.5±0.254% inhibition) and glutathione reductase (0.75±0.006% inhibition) at 8 mg/L; peroxidase (3.137±0.009 U), catalase (77.35±0.152 U), and ascorbate peroxidase (0.43±0.006 mM/mg FW) at 10 mg/L. HPLC analysis confirmed the novel presence of lupeol, an anticancer compound, in regenerated roots. These findings demonstrate the potential of CuO NPs for enhancing therapeutic metabolite production in <i>A. maurorum</i> tissue culture while suggesting optimal concentration ranges (8-10 mg/L) for maximum benefits. Further research is necessary to elucidate the molecular mechanisms governing nanoparticle-plant interactions and to address potential health implications.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5221 - 5248"},"PeriodicalIF":3.3,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144148871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}