Biotechnology Reports最新文献

{"title":"Scale-down optimization of a robust, parallelizable human induced pluripotent stem cell bioprocess for high-throughput research","authors":"James Colter ,&nbsp;Tiffany Dang ,&nbsp;Julia Malinovska ,&nbsp;Jessica May Corpuz ,&nbsp;Dora Modrcin ,&nbsp;Roman Krawetz ,&nbsp;Kartikeya Murari ,&nbsp;Michael Scott Kallos","doi":"10.1016/j.btre.2025.e00900","DOIUrl":"10.1016/j.btre.2025.e00900","url":null,"abstract":"<div><div>Human induced pluripotent stem cell (hiPSC) derived therapeutics require clinically relevant quantities of high-quality cell populations for applications in regenerative medicine. The lack of efficacy exhibited across clinical trials suggests deeper understanding of the networks governing phenotype is needed. Further, costs limit study throughput in characterizing the artificial niche relative to outcomes. We present herein an optimized strategy to enable high-throughput hiPSC expansion at &lt;20 mL research scale. We assessed viability of single cell inoculation and aggregate preformation to facilitate proliferation. We modeled aggregate characteristics against agitation rate. Our results demonstrate tunable control with fold expansion comparable to commercial systems. Marker quantification and teratoma assay confirm functional pluripotency. This approach constitutes a scalable protocol to accelerate hiPSC research, and a significant step in advancing the rate of progress in elucidating links to derivative functionality. This work will enable statistically rigorous studies targeting hiPSC and downstream phenotype for clinical manufacturing.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00900"},"PeriodicalIF":0.0,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144168992","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":"Cultivation of Cupriavidus necatorstrains on hydrolyzed lignocellulosic feedstocks widely available in Europe","authors":"Halima Aliyu Alhafiz ,&nbsp;Karin Longus ,&nbsp;Rob A.J. Verlinden ,&nbsp;Vera Lambauer ,&nbsp;Andreas Kruschitz ,&nbsp;Regina Kratzer","doi":"10.1016/j.btre.2025.e00899","DOIUrl":"10.1016/j.btre.2025.e00899","url":null,"abstract":"<div><div>Today, 85 % of the carbon in organic chemicals and their derivatives comes from fossil sources. Replacing fossil-based materials with sustainable sources requires large quantities of feedstocks and mature technologies. Biorefineries based on lignocellulose have great potential to replace fossil raw materials in the short and medium term. Here we want to pave the way for the bacterium <em>Cupriavidus necator</em> as a versatile biotechnological workhorse in future biorefineries. Wheat straw, beech, pine and spruce reflect lignocellulosic biomass from the agricultural waste and wood sectors that is widespread in Europe. Miscanthus was chosen as an emerging energy crop. Lignocellulose feedstocks were pretreated by steam explosion under variable conditions prior to enzymatic hydrolysis. Native <em>Cupriavidus necator</em> and a strain adapted by laboratory evolution were shown to grow on 16 filtered lignocellulosic hydrolysates as the sole carbon source and without prior detoxification.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00899"},"PeriodicalIF":0.0,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144240520","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":"Induction conditions that promote the effect of glycerol on recombinant protein production in Escherichia coli","authors":"Yoshihiro Ojima ,&nbsp;Hajime Saito ,&nbsp;Shintaro Miyuki ,&nbsp;Koichi Fukunaga ,&nbsp;Terumichi Tsuboi ,&nbsp;Masayuki Azuma","doi":"10.1016/j.btre.2025.e00898","DOIUrl":"10.1016/j.btre.2025.e00898","url":null,"abstract":"<div><div>Proinsulin was expressed by <em>Escherichia coli</em> SHuffle T7 with pET system in minimal medium containing glucose (Glu medium), glucose and glycerol (GluGly medium) or glycerol (Gly medium). With 100 μM IPTG, proinsulin production did not increase with glycerol. In contrast, proinsulin production per medium volume in GluGly and Gly media was approximately 3∼4-fold higher than in Glu medium with 10 μM IPTG. mRNA expression of target protein was higher in GluGly versus Glu medium, indicating that proinsulin production was enhanced by the release of glucose-induced catabolite inhibition. Although proinsulin production did not differ between GluGly and Gly media at 25 h, substrate was consumed quickly in GluGly medium with 1.55±0.12 times higher proinsulin production at 15 h. Productivity, considering the production period, was highest in the GluGly medium. This study shows a mixture of glucose and glycerol is valuable for protein production in <em>E. coli</em> with low IPTG concentrations.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00898"},"PeriodicalIF":0.0,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124178","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":"Production and evaluation of rabies immunoglobulin extracted from chicken egg yolk","authors":"Vichununt Kerdput ,&nbsp;Vararut Yodkamol ,&nbsp;Mattika Sookprasong ,&nbsp;Wongsakorn Wongwadhunyoo ,&nbsp;Iyacoob Khunsri ,&nbsp;Promsin Masrinoul ,&nbsp;Trong Wisedchanwet ,&nbsp;Naphatsamon Uthailak ,&nbsp;Yanin Limpanont ,&nbsp;Onrapak Reamtong ,&nbsp;Poom Adisakwattana ,&nbsp;Charin Thawornkuno","doi":"10.1016/j.btre.2025.e00897","DOIUrl":"10.1016/j.btre.2025.e00897","url":null,"abstract":"<div><div>This study investigated the potential of using laying hens to produce rabies immunoglobulin (RIG) due to concerns about an African Horse Sickness outbreak in 2020. The hens were immunized with rabies vaccine, resulting in the production of Immunoglobulin Y (IgY) antibodies in their egg yolks. The study found that using polyethylene glycol 6000 (PEG) was the most effective method for extracting anti-rabies IgY from egg yolks in terms of amount (1.87 mg of IgY from one mL of egg yolk), purity (83 %), and cost-effectiveness. Further purification through affinity chromatography increased the purity to 95 %, as confirmed by 1D GeLC-MS/MS, showing a purity of up to 98 %. In addition, IgY extracted using PEG demonstrated the highest neutralizing activity compared to the caprylic acid and ammonium sulfate methods. It was estimated that only 8 eggs are needed to produce a vial of RIG with the same potency as equine RIG (1000 IU/vial).</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00897"},"PeriodicalIF":0.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143947599","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":"Biogenic copper and copper oxide nanoparticles to combat multidrug-resistant Staphylococcus aureus: Green synthesis, mechanisms, resistance, and future perspectives","authors":"Gamal M. El-Sherbiny,&nbsp;M.E. Shehata,&nbsp;Mohamed H. Kalaba","doi":"10.1016/j.btre.2025.e00896","DOIUrl":"10.1016/j.btre.2025.e00896","url":null,"abstract":"<div><div>Antimicrobial resistance has increased alarmingly in recent years, with the World Health Organization identifying multidrug-resistant <em>Staphylococcus aureus</em> as a particular threat to global public health due to its extensive resistance profile and associated high mortality rates. While various metal nanoparticles have been explored as antimicrobial agents, the specific advantages of biosynthesized copper nanoparticles against MDR <em>S. aureus</em> remain inadequately consolidated in the literature.</div></div><div><h3>Objective</h3><div>This review uniquely evaluates the emerging evidence for biosynthesized copper nanoparticles as a sustainable, cost-effective, and potentially alternative to conventional antibiotics against multidrug-resistant <em>S. aureus</em> strains.</div></div><div><h3>Methods</h3><div>We systematically analyzed current literature on green synthesis methods for copper and copper oxide nanoparticles, their characterization techniques, antimicrobial mechanisms, and efficacy against multidrug-resistant <em>S. aureus</em>, focusing on identifying knowledge gaps and future research directions.</div></div><div><h3>Results</h3><div>Unlike other metal nanoparticles, biosynthesized copper nanoparticles demonstrate significant antibacterial activity against multidrug-resistant <em>S. aureus</em> through multiple simultaneous mechanisms that bacteria try to develop resistance against. Their unique physicochemical properties enable enhanced bacterial elimination compared to conventional antibiotics and other metal nanoparticles, with minimal toxicity to mammalian cells at therapeutic concentrations. Our analysis further reveals the considerable potential of these nanoparticles to overcome existing biological barriers in infection sites that limit conventional therapies.</div></div><div><h3>Conclusion</h3><div>This broad assessment of biosynthesized copper nanoparticles shows strong potential as a therapy against MDR <em>S. aureus</em> and provides a foundation for future research to address antimicrobial resistance where current treatments fail.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00896"},"PeriodicalIF":0.0,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143943270","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":"Reduction of microbial contamination and biogenic amines formation in Ras cheese using Chlorella vulgaris extracts","authors":"Diaa A. Marrez","doi":"10.1016/j.btre.2025.e00895","DOIUrl":"10.1016/j.btre.2025.e00895","url":null,"abstract":"<div><div>The present study aimed to explore the impact of <em>C. vulgaris</em> aqueous and diethyl ether extracts on reducing microbial contamination and biogenic amines (BAs) formation in Ras cheese during ripening process. Phenolic profile of aqueous extract was quantified by high performance liquid chromatography (HPLC) and compounds in ether extract were determined using gas chromatography-mass spectrometry (GC–MS). Eleven compounds were detected in aqueous extract and catechin was the major (54.82 µg/g), while 14 compounds were detected in ether extract of which the main compounds hexadecane (14.26 %) and 9, 12-Octadecadienoic acid, methyl ester (9.61 %). <em>C. vulgaris</em> aqueous and ether extracts observed antibacterial activity against seven strains of foodborne bacteria with minimum inhibitory concentration (MIC) ranged from 0.33 to 1.43 mg/mL and antifungal activity against 9 strains of toxigenic fungi with MIC values from 0.42 to 1.74 mg/mL. Ras cheese treated with ether extract had the highest reduction in formation of tryptamine (93.4 %), β-phenylethylamine (86.4 %), Putrescine (88.3 %), Cadaverine (81.8 %), Histamine (62.9 %), Serotonin (92.1 %), Tyramine (87.2 %), Spermidine (80.8) and Spermine (86.8 %) compared to cheese treated with aqueous extract and control samples. Also, the microbial load (total bacterial count, yeasts and molds count, proteolytic bacterial count) in Ras cheese samples treated with <em>C. vulgaris</em> aqueous and diethyl ether extracts were lower than the control sample. Whereas, no growth of coliform group, Staphylococci and Salmonella were detected in treated samples and control. The microalga <em>C. vulgaris</em> extracts considered promising source from natural ingredients to reduce biogenic amines content and microbial load in cheese manufacturing.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00895"},"PeriodicalIF":0.0,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143895659","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":"Unveiling the potential biochemical effects of selected heterocyclic compounds as human Type-A γ-aminobutyric acid (GABA A) Modulator: An Insilico Approach","authors":"Abel Kolawole Oyebamiji ,&nbsp;Sunday Adewale Akintelu ,&nbsp;Oluwakemi Ebenezer ,&nbsp;Faith Eniola Olujinmi ,&nbsp;David O. Adekunle ,&nbsp;Adesoji Alani Olanrewaju ,&nbsp;Omowumi Temitayo Akinola ,&nbsp;Samson Olusegun Afolabi ,&nbsp;Ehimen Anastasia Erazua ,&nbsp;Ayodeji Arnold Olaseinde","doi":"10.1016/j.btre.2025.e00894","DOIUrl":"10.1016/j.btre.2025.e00894","url":null,"abstract":"<div><h3>Background</h3><div>Investigating the bioactivities of zuranolone derivatives as Type-A γ-aminobutyric acid inhibitors which will thereby down-regulate postpartum depression is considered a crucial study.</div></div><div><h3>Method</h3><div>This study is aimed at investigating the biochemical activities of 1-(2-((3R,5R,8R,9R,10S,13S,14S,17S)-3‑hydroxy-3,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-oxoethyl)-1H-pyrazole-4-carbonitrile derivatives against type-A γ-aminobutyric acid (GABA A) which will thereby enhance the activity of γ-aminobutyric acid (GABA) in human central nervous system.</div></div><div><h3>Results</h3><div>In this work, series of computational tools such as Spartan 14, molecular operating environment software, Gromacs and Admetsar1 were explored and the studied compounds were subjected to this software which resulted to series of results. Vacuum was observed to have highest influence on highest occupied molecular orbital (E_H) of compound 1 and water as well as ethanol reduces its ability to donate electron to the nearby molecules. Also, the effect of water and ethanol were investigated on the studied compound via lowest unoccupied molecular orbital (E_L) and energy gap and the results were reported appropriately. The molecular docking investigation was carried out on the studied compounds and Type-A γ-aminobutyric acid (pdb id: 4cof) and the compounds 3 with calculated binding affinity value of -7.32433319kcal/mol as well as pi-H as the non-bonding interaction were observed which therefore confirm the potential ability of compound to inhibit the target than other studied compound. Also, compound 1 and reference compound were subjected to molecular dynamic simulation study and the actual binding energy for the selected compounds were obtained and reported.</div></div><div><h3>Conclusion</h3><div>Our findings from this work may open door for the design of several 1-(2-((3R,5R,8R,9R,10S,13S,14S,17S)-3-(benzyloxy)-3,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-oxoethyl)-1H-pyrazole-4-carbonitrile derivatives as potential Type-A γ-aminobutyric acid inhibitors.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00894"},"PeriodicalIF":0.0,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848379","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":"Rhizobial, passenger nodule endophytes and phyllosphere bacteria in combination with acyl homoserine lactones enhances the growth and yield of groundnut","authors":"Sivakumar Madhan ,&nbsp;Yuvasri Errakutty Arunan ,&nbsp;Anandham Rangasamy ,&nbsp;Balachandar Dananjeyan ,&nbsp;Johnson Iruthayasamy ,&nbsp;Manimaran Gajendiran ,&nbsp;Krishnamoorthy Ramasamy ,&nbsp;Raghu Rajasekaran ,&nbsp;Vincent Saminathan","doi":"10.1016/j.btre.2025.e00893","DOIUrl":"10.1016/j.btre.2025.e00893","url":null,"abstract":"<div><div>Quorum sensing (QS) mechanisms play an essential role in mediating several signals and plant-bacteria interactions, promoting plant growth. This study demonstrated production of multiple Homoserine lactone (HSL) molecules like C6 HSL, C7 HSL, C8 HSL, 3-Hydroxy-C8-HSL and 3-oxo-C14 HSL in rhizobial and passenger endophytes and phyllospheric bacteria which regulated production of plant growth promoting traits <em>viz.,</em> indole acetic acid and exo-polysaccharide production, biofilm formation, and motility. Quorum quenching (QQ) molecules like salicylic acid, gallic acid, and disalicylic acid impaired these traits, but exogenous addition of QS molecules (C7HSL and 3-oxo-C14 HSL) restored these inhibitory effects of QQ compounds. The pot culture experiment revealed that the treatment involving <em>Methylobacterium populi</em> TMV7–4 or <em>Enterobacter cloacae</em> S23 with salicylic acid, C7HSL and 3-oxo-C14 HSL significantly enhanced plant growth including root length, nodulation, pod formation, soil available nutrients and plant nutrients uptake. In future field validation is required for the use of QS molecules in improving groundnut production.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00893"},"PeriodicalIF":0.0,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143823748","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-situ partial reduction of biochar by overlaying a syngas stream","authors":"Valentin Chataigner ,&nbsp;Dominique Tarlet ,&nbsp;François Ricoul ,&nbsp;Jérôme Bellettre","doi":"10.1016/j.btre.2025.e00892","DOIUrl":"10.1016/j.btre.2025.e00892","url":null,"abstract":"<div><div>This work focuses on the partial reduction of a hot biochar by a hot syngas coming out of the partial oxidation zone in the Ariane® pyrolysis prototype. The patented geometry of this pyrolyser, with separate pyrolysis and partial oxidation zones, enables easy management of biochar and syngas flows. The aim is to increase the specific surface area of the biochar to achieve more agronomic specifications, while maintaining a high carbon content in the biochar. The experimental implementation of this partial reduction is achieved by adding a partial reduction zone at the bottom of the reactor, enabling direct contact between biochar and syngas. This part now enables biochar and syngas streams to be mixed in a high-temperature zone (<span><math><mrow><mo>&gt;</mo><msup><mn>500</mn><mo>∘</mo></msup><mi>C</mi></mrow></math></span>). The addition of this part increased the specific surface area of the biochar by <span><math><mrow><mn>250</mn><mspace></mspace><msup><mrow><mi>m</mi></mrow><mn>2</mn></msup><mo>/</mo><mi>g</mi></mrow></math></span> and preserved the carbon content of the biochar regardless of the biomass flow rate. Around <span><math><mrow><mn>20</mn><mspace></mspace><mo>%</mo></mrow></math></span> of the carbon in the biochar is consumed by the reduction and transferred from the biochar to the syngas. The syngas is thus enriched in particular in <span><math><msub><mi>H</mi><mn>2</mn></msub></math></span> and <span><math><mrow><mi>C</mi><msub><mi>O</mi><mn>2</mn></msub></mrow></math></span> and depleted in <span><math><mrow><mi>C</mi><msub><mi>H</mi><mn>4</mn></msub></mrow></math></span>, which opens up a wider range of development opportunities for this gas (biomethane, biohydrogen, biokerosene, cogeneration, etc.). Thanks to this activation method, a better compromise has been achieved (compared with the previous version of the prototype) between quality (specific surface area and carbon content) and quantity of biochar produced (anhydrous yield).</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00892"},"PeriodicalIF":0.0,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143783393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Montbretia flowers as a source of bioactive crocins: Biotechnology tools and delivery systems","authors":"Lucía Morote ,&nbsp;Cristian Martínez Martínez Fajardo ,&nbsp;María Mondéjar López ,&nbsp;Elena Moreno-Gimenez ,&nbsp;Ángela Rubio-Moraga ,&nbsp;Olivia Costantina Demurtas ,&nbsp;Gianfranco Diretto ,&nbsp;Enrique Niza ,&nbsp;Verónica Aragonés ,&nbsp;Alberto López Jiménez ,&nbsp;José-Antonio Daròs ,&nbsp;Oussama Ahrazem ,&nbsp;Lourdes Gómez-Gómez","doi":"10.1016/j.btre.2025.e00891","DOIUrl":"10.1016/j.btre.2025.e00891","url":null,"abstract":"<div><div>Crocins are potent antioxidants with significant therapeutic potential, exhibiting anti-inflammatory, anticancer, and antidepressant properties. The ornamental plant <em>Crocos-mia x crocosmiiflora</em> is widely cultivated for its aromatic and vibrant flowers. In this study, we identified unique crocins as the primary pigments responsible for the flower's coloration. These metabolites predominantly consist of molecules with eight glucose units, followed by crocins containing six and seven glucose units. To elucidate the mo-lecular mechanisms underlying crocin biosynthesis in <em>C. x crocosmiiflora</em>, tran-scriptomic analysis was performed to identify key carotenoid cleavage dioxygenase (CCD) genes. Using <em>Crocus sativus</em> CsCCD2L gene as a bait, we identified a CCD transcript from the transcriptome data. Phylogenetic analysis revealed that the identified CCD belongs to the CCD2 subfamily, and it was designated as CroCCD2. Functional characterization of CroCCD2 was carried out using bacterial expression systems and <em>Nicotiana benthamiana</em> plants with a virus-mediated expression system. These experiments demonstrated that CroCCD2 efficiently converts the precursor zeaxanthin into crocetin, a key intermediate in crocin biosynthesis. Furthermore, we investigated the bioactivity of crocins and discovered that their anti-inflammatory effects depend on their vehiculation within exosomes or liposomes. This suggests that the transport mechanism is critical for the biological activity of crocins. Our findings highlight the specialization of CCD subfamilies in monocots and dicots for crocin biosynthesis and provide evidence of the anti-inflammatory activity of exosome-transported crocins. This study establishes a foundation for further research into the metabolic network of crocins in <em>C. x crocosmiiflora</em> and suggests that the CroCCD2 gene could be introduced into other crop plants to produce these bioactive apocarotenoids.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00891"},"PeriodicalIF":0.0,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143738734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}