Amanda Lillberg, Maria Pajumo, Eevi Haajanen, Terhi Viinikanoja, Anneli Ritala
{"title":"Valorization of shopping center side- and waste streams with plant cell culture technology – Prospects and cost reduction","authors":"Amanda Lillberg, Maria Pajumo, Eevi Haajanen, Terhi Viinikanoja, Anneli Ritala","doi":"10.1016/j.jbiotec.2025.05.006","DOIUrl":"10.1016/j.jbiotec.2025.05.006","url":null,"abstract":"<div><div>Upcycling potential of side-streams and waste material, originating from a shopping center were studied as a feedstock for plant cell cultures of arctic bramble (<em>Rubus arcticus</em> L.), barley (<em>Hordeum vulgare</em> L.) and tobacco BY-2 (<em>Nicotiana tabacum</em> L.). Soft drink waste mix, orange peels, expired bread, spent coffee grounds, and brewer’s spent grain, were used in cultivation experiments to supplement sucrose in the basic growth medium. Artificial urine was included to represent a future scenario, where also human urine could be used as a circular feedstock along improvement in purification technologies allowing removal of microbes and toxic molecules. Diluted artificial urine was used to replace elemental macronutrients e.g., ammonium, phosphate, and sulphate in the culture medium. Soft drink waste mix and artificial urine contributed to the highest biomass accumulation of the tested side streams. Culture medium composition optimization, exploiting combination of soft drink waste mix and artificial urine, was conducted with design of experiments software. The optimized culture medium contributed to notable culture medium cost reductions of 55 %, 33 % and 45 % for arctic bramble, tobacco BY-2 and barley cell lines, respectively, without compromising the biomass yield. However, modified culture medium induced some changes in the amino acid composition of arctic bramble plant cell biomass, e.g., increased asparagine and decreased arginine, alanine and tyrosine contents. Thus, attention should be paid to the amino acid composition in further studies. Our findings confirm the potential of upcycling side streams and replacing costly growth medium components with so far unutilized resources.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 150-158"},"PeriodicalIF":4.1,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144016279","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Luona Ye, Zitang Xu, Yunchong Li, Pengbo Wang, Yunjun Yan, Jinyong Yan
{"title":"A chitosan-integrated antibacterial protein composite nanocomplex derived from barnacle cement and spider silk","authors":"Luona Ye, Zitang Xu, Yunchong Li, Pengbo Wang, Yunjun Yan, Jinyong Yan","doi":"10.1016/j.jbiotec.2025.05.007","DOIUrl":"10.1016/j.jbiotec.2025.05.007","url":null,"abstract":"<div><div>While barnacle cement protein cp19k (from <em>Megabalanus rosa</em>) possesses remarkable adhesion properties and spider silk protein MaSp1 (from <em>Nephila clavata</em> dragline silk) demonstrates exceptional toughness, their advancements in medical biomaterials are significantly hindered by their limitations in antimicrobial properties. In this study, composite nanocomplexes incorporating chitosan and proteins derived from barnacle cement and spider silk were designed and biofabricated for enhanced antibacterial properties. The impact of chitosan’s molecular weight on the properties of nanocomplexes comprising cp19k-MaSp1/chitosan, MaSp1/chitosan, and cp19k/chitosan was evaluated. The results revealed that low molecular weight chitosan (LMWC, Mw = 1 kDa) forms nanocomplexes that exhibit distinct structural differences in comparison to those formed with high molecular weight chitosan (HMWC, Mw ≥ 150 kDa). Furthermore, cp19k-MaSp1/C<sub>150k</sub> exhibited the most potent antibacterial activity against <em>E. coli</em> and <em>S. aureus</em>, surpassing the performance of cp19k, MaSp1, cp19k-MaSp1, and chitosan individually, achieving inhibition by disrupting the bacterial cell membrane structure and elevating the intracellular ROS level. Meanwhile, On day 6, the viability of HUVECs (Human Umbilical Vein Endothelial Cells) of cp19k-MaSp1/C<sub>150k</sub> had attained a level of 145.21 ± 6.23 %, representing a substantial elevation when compared to C<sub>150k</sub>. The remarkable biocompatibility of nanocomplexes cp19k-MaSp1/C<sub>150k</sub> holds potential for application in wound dressings and tissue repair.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 57-71"},"PeriodicalIF":4.1,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143931888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Interferon gamma-induced hub genes and key pathways: A study based on biological network analysis and experimental validation","authors":"Reza Heidari-Japelaghi , Mostafa Valizadeh , Raheem Haddad","doi":"10.1016/j.jbiotec.2025.04.022","DOIUrl":"10.1016/j.jbiotec.2025.04.022","url":null,"abstract":"<div><div>By performing a biological network analysis, we identified some hub genes, which were up- or down-regulated in the breast cancer (BC) cell line after treatment with IFN-γ. Moreover, several pathways including cytokine-cytokine receptor interaction, TNF signaling pathway, NOD-like receptor signaling pathway, and NF-κB signaling pathway were detected that their activation leads to the antiproliferation, proapoptosis, and antiviral activities. To validate <em>in silico</em> results, the bioactivity of recombinant human IFN-γ (hIFN-γ) produced in different hosts was analyzed by antiviral and anticancer assays. The antiviral role of the hIFN-γ preparations was evaluated by inhibition of Vesicular Stomatitis Virus (VSV)-mediated cytopathic effects on Vero cells. A dose-dependent increase in cell viability was observed at different concentrations of recombinant proteins. The maximum amount of the cell viability detected for the hIFN-γ preparations was determined at a concentration of 32.00 pg/mL. To analyze the cytotoxic efficacy of the hIFN-γ preparations on the growth and development of tumor cells, a BC cell line (MCF-7) was treated with both recombinant protein forms in a time- and dose-dependent way. The highest level of inhibiting cell proliferation was detected at a concentration of 32.00 pg/mL hIFN-γ after 72 h incubation. Anticancer and antiviral functions of IFN-γ were confirmed via the expression analysis of hub genes <em>cd74</em>, <em>cxcl10</em>, <em>il6</em>, and <em>stat1</em> using RT-PCR. Furthermore, the hIFN-γ preparations were significantly able to up-regulate the expression of proapoptotic <em>Bax</em> and <em>p53</em> and to down-regulate <em>Bcl-2</em> as an antiapoptotic gene, showing the cytotoxic effect of hIFN-γ toward MCF-7 cells via apoptosis induction.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 72-87"},"PeriodicalIF":4.1,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143935541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yating Zhang , Xixi Wang , Dong Su , Ling Zhao , Kailiang Leng , Junkui Miao , Yueqin Yu
{"title":"Effective astaxanthin production from flocculated Haematococcus pluvialis via biofilm cultivation in a tri-layer tray bioreactor","authors":"Yating Zhang , Xixi Wang , Dong Su , Ling Zhao , Kailiang Leng , Junkui Miao , Yueqin Yu","doi":"10.1016/j.jbiotec.2025.05.002","DOIUrl":"10.1016/j.jbiotec.2025.05.002","url":null,"abstract":"<div><div><em>Haematococcus pluvialis</em>, renowned for its high astaxanthin content, is a prime candidate for commercial-scale production of natural astaxanthin. While biofilm cultivation shows potential for enhancing astaxanthin accumulation in <em>H. pluvialis</em>, it faces challenges in effectively harvesting green-vegetative <em>H. pluvialis</em> and mitigating the \"dark zone\" issue within the biofilm. Our study demonstrated that flocculation using a combination of chitosan and NaOH was an effective method for harvesting green-vegetative <em>H. pluvialis</em>, which surpasses centrifugation in both biomass and astaxanthin production. A tri-layer tray bioreactor was developed to enhance astaxanthin production via biofilm redistribution by using a rinsing method, and its effectiveness was further supported by a tray photobioreactor with an integrated automatic rinsing system. The biofilm method achieved an optimal specific light energy consumption for astaxanthin production of 9.42 kWh g<sup>−1</sup> at the light intensity of 150 μmol m<sup>−2</sup> s<sup>−1</sup>, presenting its potential for commercial-scale cultivation of <em>H. pluvialis</em> for astaxanthin production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 88-98"},"PeriodicalIF":4.1,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143935535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Anna Bechtel, Lucas Kettner, Marie Jenz, Daniel Mopils, Lutz Fischer
{"title":"Recombinant production of a diamine oxidase in Komagataella phaffii and in vitro degradation of histamine in simulated intestinal fluid","authors":"Anna Bechtel, Lucas Kettner, Marie Jenz, Daniel Mopils, Lutz Fischer","doi":"10.1016/j.jbiotec.2025.05.003","DOIUrl":"10.1016/j.jbiotec.2025.05.003","url":null,"abstract":"<div><div>Diamine oxidase (DAO-1) from <em>Yarrowia lipolytica</em> was recombinantly produced in <em>Komagataella phaffii</em> in fed-batch bioreactor cultivations using the constitutive <em>GAP</em> and methanol-inducible <em>AOX1</em> promoters. Utilizing an exponential glucose feed, a maximum DAO-1 activity of 43 ± 7 µkat/L<sub>culture</sub> was achieved after 37 h of cultivation using the <em>GAP</em> promoter. When the <em>AOX1</em> promoter was used, a maximum DAO-1 activity of 230 ± 13 µkat/L<sub>culture</sub> was achieved after 90 h of cultivation with a stepwise increase in the methanol feed. Thus, approximately 15- (<em>GAP</em> promoter) and 82-fold (<em>AOX1</em> promoter) higher DAO-1 activities were obtained in <em>K. phaffii</em> compared to homologous recombinant production in <em>Y. lipolytica</em>. The DAO-1 produced in <em>K. phaffii</em> has been partially purified and was formulated as a sucrose-based tablet containing 4500 nkat DAO-1 activity. The DAO-1 tablet was tested for histamine degradation in a simulated intestinal fluid in the presence of food components, and approximately 90 % of the histamine applied initially (75 mg) was degraded in 90 minutes. This is the first time that a DAO-1 tablet has been able to break down such large amounts of histamine under simulated intestinal fluid conditions, which could help people with histamine intolerance in the future.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 39-47"},"PeriodicalIF":4.1,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143927527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Edmilson Clarindo de Siqueira , Aline de Andrade Alves , Maria Paloma Silva de Barros , Rayane da Silva Vale , Páblo Eugênio da Costa e Silva , Jonas Contiero , Emmanuel Damilano Dutra , Laureen Michelle Houllou
{"title":"Integrated production of polyhydroxyalkanoates and rhamnolipids: Insights in cultivation conditions and metabolic engineering","authors":"Edmilson Clarindo de Siqueira , Aline de Andrade Alves , Maria Paloma Silva de Barros , Rayane da Silva Vale , Páblo Eugênio da Costa e Silva , Jonas Contiero , Emmanuel Damilano Dutra , Laureen Michelle Houllou","doi":"10.1016/j.jbiotec.2025.05.001","DOIUrl":"10.1016/j.jbiotec.2025.05.001","url":null,"abstract":"<div><div>Polyhydroxyalkanoates and biosurfactants have the potential to play a significant role in emerging bioeconomic chains. With growing environmental worries about the excessive consumption of fossil fuel derivatives, significant focus has been paid to a renewable-based economy known as the circular bioeconomy. Polyhydroxyalkanoates (PHAs) are a type of biodegradable, hydrophobic, non-toxic, thermoplastic polymer created by microbial processes that have good physicochemical properties. Rhamnolipids (RhL) are amphipathic, biodegradable, and biocompatible compounds with outstanding emulsification capabilities. Unfortunately, commercial manufacturing of PHA and RhL remains limited due to their high production costs as compared to standard polymers and surfactants. The combined manufacture of PHA and RhL can lower production costs and is an ideal option for creating two widely applicable commodities on the market. This work provides a general overview of PHA and RhL co-production, focusing on the use of renewable materials and important aspects that are directly related to cultivation conditions, as well as genetic and metabolic engineering strategies to optimize PHA and RhL production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 17-25"},"PeriodicalIF":4.1,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143922354","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Matheus Pereira de Carvalho , Michele Greque de Morais , Lucielen Oliveira dos Santos , Céline Laroche , Jorge Alberto Vieira Costa
{"title":"Production of biomass and biomolecules in Limnospira indica PCC 8005 cultivation under magnetic fields and polymeric nanofibers","authors":"Matheus Pereira de Carvalho , Michele Greque de Morais , Lucielen Oliveira dos Santos , Céline Laroche , Jorge Alberto Vieira Costa","doi":"10.1016/j.jbiotec.2025.05.004","DOIUrl":"10.1016/j.jbiotec.2025.05.004","url":null,"abstract":"<div><div>Researchers often apply physical, chemical, or biological stresses to cyanobacteria cultivation to enhance biomass production by triggering cellular adaptation mechanisms, increasing growth or boosting target compound synthesis. Static magnetic fields (SMF) offer a non-toxic, cost-effective way to modulate microalgal growth, alter biomass composition, and promote metabolite production. Polymeric nanofibers (Nano) function as a physical barrier in cultivation, while monoethanolamine (MEA) acts as a chemical absorbent, reducing CO₂ loss and enhancing biofixation. This study investigated the effects of SMF and nanofibers on the biomass yield and molecular composition of <em>Limnospira indica</em> PCC 8005. The combined SMF and Nano treatment achieved the highest biomass yield (5.87 ± 0.06 g L⁻¹), a 28 % increase compared to the control. SMF application increased protein content by 16 % but reduced carbohydrate levels by 73 % relative to the nanofiber-only treatment (39.58 ± 0.98 % ww⁻¹). Exopolysaccharide (EPS) produced under the SMF+NanoMEA treatment contained 39.9 % uronic acid, while the Nano-only treatment had the highest sulphate content (8.4 %) but the lowest uronic acid concentration (25.4 %). The EPS were identified as acidic, sulphated polysaccharides. SMF and nanofibers significantly enhances biomass production, alters the carbohydrate and protein proportions in biomass, and influences the composition of sugars, acids, and sulphate in exopolysaccharides.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 48-56"},"PeriodicalIF":4.1,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143931889","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Biocatalytic production of a monoamine oxidase B/catechol-O-methyltransferase inhibitor from piperine by engineered P450 BM3.","authors":"Mariusz Brzoski , Samuele Irudal , Elena Gazzano , Roberto Buscaino , Guido Viscardi , Giovanna Di Nardo , Gianfranco Gilardi","doi":"10.1016/j.jbiotec.2025.04.024","DOIUrl":"10.1016/j.jbiotec.2025.04.024","url":null,"abstract":"<div><div>The single-step biotransformation of the natural compound piperine into a known dual inhibitor of monoamine oxidase B (MAO-B) and catechol-O-methyltransferase (COMT), was achieved by cytochrome P450 BM3 wild-type and the D251G/Q307H double mutant. This compound is used for research in neurodegenerative disorders, such as Parkinson’s disease, and its value in the market is ∼14,000 €/g. Currently, it is produced by chemical synthesis requiring incubation of piperine with boron tribromide (BBr<sub>3</sub>) in dichloromethane with yield of product not exceeding 55 % and using tedious and long procedure for its production and isolation. The P450 D251G/Q307H double mutant exhibited a 3-fold increase in catalytic efficiency compared to the wild-type enzyme, achieving high conversion (51.6 % of conversion in 15 minutes) under mild, environmentally friendly conditions. The yield of production was 0.01 mg of the inhibitor in 1 mL of reaction in 15 minutes at 28°C using the purified enzyme. Moreover, biological assays demonstrated that the resulting compound has a novel and stronger antioxidant and antimicrobial activities, respectively, when compared to piperine. The data further demonstrates the broader potential of engineered enzymes as versatile and sustainable tools in industrial biotechnology, offering an efficient platform for the modification of natural compounds to produce bioactive molecules.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 8-16"},"PeriodicalIF":4.1,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143917681","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Fermentative production of α-ketoisovalerate and α-ketoisocaproate by metabolically engineered Corynebacterium glutamicum","authors":"Yanan Wang, Weixuan Dong, Yulong Gao, Jiaxiang Kuang, Xinyu Zhou, Feiao Wang, Siyu Tian, Yanjun Li","doi":"10.1016/j.jbiotec.2025.04.023","DOIUrl":"10.1016/j.jbiotec.2025.04.023","url":null,"abstract":"<div><div>Alpha-ketoisovalerate (KIV) and α-ketoisocaproate (KIC) are widely used as food additives and in the synthesis of pharmaceuticals and higher alcohols. Current chemical synthesis methods are environmentally harmful, and whole-cell catalysis processes are costly due to expensive substrates. Direct fermentative production of KIV and KIC from glucose is a promising alternative, although research in this area remains limited. In this study, we engineered an L-valine-overproducing <em>Corynebacterium glutamicum</em> strain for KIV and KIC production. We inactivated leucine dehydrogenase and isopropylmalate synthase to block the formation of L-valine and KIC, resulting in the production of 53.5 g/L KIV with a yield of 0.16 g/g glucose and a productivity of 0.70 g/L·h⁻¹ in a 5-L fermentor. Next, we overexpressed genes in the L-leucine biosynthesis pathway (<em>leuA</em>, <em>leuCD</em>, and <em>leuB</em>) by introducing a feedback-resistant <em>leuA</em> (<em>leuA</em><sup>fbr</sup>) in a plasmid-based system, deleting the transcriptional repressor gene <em>ltbR</em>, and increasing the gene copy numbers of <em>leuCD</em> and <em>leuB</em> under a strong promoter, creating a high-KIC-producing strain. Acetate supplementation enhanced acetyl-CoA supply, increasing KIC production while reducing KIV accumulation. The final strain produced 79.8 g/L KIC with a yield of 0.29 g/g glucose and a productivity of 1.05 g/L·h⁻¹ in a 5-L fermentor, surpassing previous fermentation results and most whole-cell catalysis processes, highlighting its industrial application potential.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"405 ","pages":"Pages 1-7"},"PeriodicalIF":4.1,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143922353","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Fabrication of a sensitive neurotransmitter detecting amperometric biosensor employing laccase nanoparticles on a pencil graphite electrode","authors":"Himani Guliya , Suman Lata , Reeti Chaudhary","doi":"10.1016/j.jbiotec.2025.04.021","DOIUrl":"10.1016/j.jbiotec.2025.04.021","url":null,"abstract":"<div><div>Neurotransmitters including dopamine, adrenaline, and noradrenaline are members of the important class of biogenic amines known as catecholamines. They perform crucial roles in various physiological processes and are frequently associated with stress responses, neurodegenerative and cardiovascular diseases, including Parkinson's and Alzheimer's. This research presents the fabrication of a novel amperometric biosensor designed to detect catecholamine levels with high specificity and sensitivity. The fabrication of the biosensor is based on the immobilization of synthesized cysteine functionalized laccase nanoparticles (Lac-NPs) onto the pencil graphite electrode (PGE). The successful synthesis of Lac-NPs, along with their immobilization and the fabrication of the Lac-NPs/PGE biosensor, was validated through various techniques, including Transmission Electron Microscopy (TEM), Fourier Transform Infrared Spectrophotometer (FTIR), UV–visible Spectroscopy, Dynamic Light Scattering (DLS), Zeta potential, Scanning Electron Microscope (SEM), Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). The biosensor was optimized at various pH, temperature, scan rate and response times to ensure a high-performance sensor with rapid response times and stability for better detection. The Lac-NPs/PGE biosensor showed high sensitivity (2320.0 µA/mM cm<sup>2</sup>), a lower limit of detection (LOD) (0.12 µM), and a broad linear range (0.1–800.0 µM) with a coefficient of determination of R<sup>2</sup>= 0.999. In the analysis of real pharmaceutical samples of neurotransmitters, high recovery rates (94.0–99.0 %) have been attained. Superior analytical performance resulting from this simple fabrication process and cost-effective PGE shows this biosensor is a promising tool for the accurate and real-time monitoring of catecholamine levels, with potential applications in clinical diagnostics, neurobiology, and environmental analysis.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"404 ","pages":"Pages 152-161"},"PeriodicalIF":4.1,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143891008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}