Applied Biochemistry and Biotechnology最新文献

{"title":"Evaluation of the Effects of Di-(2-ethylhexyl) phthalate (DEHP) on Caenorhabditis elegans Survival and Fertility.","authors":"Alper Zöngür","doi":"10.1007/s12010-024-05032-z","DOIUrl":"https://doi.org/10.1007/s12010-024-05032-z","url":null,"abstract":"<p><p>Di-2-ethylhexyl (DEHP), which is widely used in industrial products, is produced annually in excess of 2 million tons worldwide. DEHP is an endocrine disruptor and one of the major environmental pollutant chemicals (EDCs) in nature. There is some information about the effects of these products, which provide great advantages in every respect, on human health and the environment. In this study, C. elegans organism was used to evaluate the health and environmental risks of DEHP. The survival and fertility effects of DEHP on the C. elegans organism were examined and the results were evaluated. In the study, it was determined that DEHP not only shortened the survival time of C. elegans but also caused a decrease in fertility. DEHP (0.625 mM and 10 mM) caused a 23.2-30.6% decrease in fertility. Additionally, the LC₅₀ (50% lethal concentration) value of DEHP was found to be 321 µg/mL.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141858689","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":"Production of Lipopeptide Biosurfactant Using Wastewater from Parboiled Paddy Rice and Evaluation of Antifungal Property of the Biosurfactant Against Two Dermatophyte Fungi.","authors":"Chandana Malakar, Mehjabin Ali, Rupshikha Patowary, Suresh Deka","doi":"10.1007/s12010-024-05000-7","DOIUrl":"https://doi.org/10.1007/s12010-024-05000-7","url":null,"abstract":"<p><p>A previously isolated lipopeptide biosurfactant-producing bacterium Bacillus licheniformis SCV1 was investigated for the production of the biosurfactant using wastewater from parboiled paddy rice. The biosurfactant thus produced was evaluated for its antifungal property against dermatophyte fungi Trichophyton ajelloi and Microsporum fulvum. Results revealed that the bacterial strain reduced surface tension of the media from 56.16 ± 1 mN/m to 35 ± 0.9 mN/m within 12 h, which further shrank to 29.3 ± 1 mN/m in 24 h of incubation. The yield of the biosurfactant was 3.15 ± 0.25 g/L at 48 h of incubation. The obtained biosurfactant exhibited efficient emulsifying activity against a wide range of hydrophobic substrates such as crude oil, olive oil, engine oil, and kerosene oil used in the study. The critical micelle concentration of the biosurfactant was found to be 80 mg/L. Structural characterization using FT-IR and TLC revealed that the biosurfactant produced by the strain in the wastewater is a lipopeptide consisting of surfactin and iturin. LCMS analysis revealed that the surfactin homologs range from C12 to C17-surfactin while the iturin contains C13 to C17-iturin homologs. It also revealed an in vitro study that the biosurfactant has antifungal properties against dermatophyte fungi Trichophyton ajelloi and Microsporum fulvum. Microscopic observation of the hyphae of the treated dermatophyte revealed disruption and fissure of the mycelia. The chemical composition of the wastewater revealed that it contains adequate nutritional composition and micronutrients to support bacterial growth. This is the first report that the wastewater of parboiled paddy could be used as a low-cost substrate for the production of lipopeptide biosurfactant, and the biosurfactant could be used for preventing dermatophytes fungi.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141858690","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":"Does Scaling Up Pig Farming Promote Carbon Neutrality Among Pig Farmers?","authors":"Wenjie Yao","doi":"10.1007/s12010-024-05029-8","DOIUrl":"https://doi.org/10.1007/s12010-024-05029-8","url":null,"abstract":"<p><p>Pig farming is a major source of carbon emissions in China's livestock industry, and the promotion and application of carbon neutral technologies dedicated to carbon reduction and sequestration is a key measure to promote the realization of carbon neutrality in pig farming industry. In the absence of a system for carbon emission reduction in agriculture, it is important to explore whether large-scale pig farming can spontaneously promote the application of carbon neutral technologies by pig farmers. Combined with the technical background of carbon neutrality in pig farming industry and using the field survey data of 468 pig farmers in Shandong Province, this paper empirically analyzed the effect of large-scale pig farming on the carbon neutral behavior of pig farmers. The findings are threefold. Firstly, because of the effect of economies of scale in the application of carbon neutral technologies, large-scale pig farming can promote the carbon neutral behavior of pig farmers. However, the effect of economies of scale in the application of carbon neutral technologies has a boundary, and the annual output of pigs should be maintained at 3000 to 4999. Secondly, among the effects of pig breeding scale on the application and its degree of composting project, state of planting and breeding cycle has no regulating effect. Thirdly, among the effects of pig breeding scale on the application and its degree of biogas engineering, both state of planting and breeding cycle and subsidy for construction of biogas digester have negative regulating effects. In this paper, the pig farming industry is taken as a typical example to reveal the endogenous incentive mechanism of promoting agricultural carbon emission reduction, which provides theoretical reference and experience enlightenment for the formulation of effective policies to promote agricultural carbon neutrality, which are different from industrial industries.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141858688","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":"Purification of Potential Antimicrobial Metabolites from Endophytic Fusarium oxysporum Isolated from Myrtus communis.","authors":"Saeed Ullah Khattak, Mansoor Ahmad, Jamshaid Ahmad, Saima Ikram, Sajjad Ahmad, Fahad M Alshabrmi, Eid A Alatawi","doi":"10.1007/s12010-024-05016-z","DOIUrl":"https://doi.org/10.1007/s12010-024-05016-z","url":null,"abstract":"<p><p>The rise of microbial resistance and emerging infections pose significant health threats. Natural products from endophytic fungi offer a promising source of novel compounds with the potential as major drug leads. This research aims to screen Myrtus communis and Moringa oleifera for endophytic fungi and screen their metabolites for antibacterial and antifungal potential. Six endophytic fungal strains were isolated using a potato dextrose agar (PDA) medium. The M. communis isolates were designated MC1, MC2, and MC3, and the M. oleifera isolates were named MO1, MO2, and MO3. Preliminary bioactivity testing revealed that the MC3 isolate exhibited significant growth inhibition against multidrug-resistant bacterial and fungal pathogens, including Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and Candida glabrata. The MC3 isolate was identified as Fusarium oxysporum through morphological and microscopic methods. For metabolite production, the fungal strain was cultured in potato dextrose broth (PDB) medium at 28 °C for 14 days in a shaking incubator. The metabolites were purified using various chromatographic techniques, HPLC and GC-MS. The GC-MS analysis of the bioactive compound containing fungal strain (F. oxysporum) revealed multiple compounds at different retention times using the NIST-20 Library. Based on RSI values and probability indices, two compounds were targeted for further purification. Structure elucidation was performed using 1D and 2D nuclear magnetic resonance (NMR) experiments on a Varian 500 NMR machine. The compounds identified were ethyl iso-allocholate (C₂₆H₄₄O₅, exact mass 436.32) and 1-monolinoleoyl glycerol trimethylsilyl ether (C₂₇H₅₆O₄Si₂, exact mass 500.37). The MS (NIST-20) library facilitated the investigation of the in silico antimicrobial activity of these compounds against the elastase virulence protein of P. aeruginosa and protease Sapp1p from C. parapsilosis. Both the compounds were docked with druggable proteins using the Glide induced fit docking (IFD) algorithm. The ethyl iso-allocholate and 1-monolinoleoyl glycerol trimethylsilyl ether compounds showed binding scores - 10.07 kcal mol^-1 and - 7.47 kcal mol^-1 against elastase, and - 8.16 kcal mol^-1 and - 6.89 kcal mol^-1 against aspartic protease, respectively. In vitro studies confirmed the inhibitory activity of these compounds against multidrug-resistant P. aeruginosa and E. faecalis. Ethyl iso-allocholate exhibited higher bioactivity against P. aeruginosa with inhibition rates of 41%, 27%, and 35% at concentrations of 1000, 500, and 250 µg mL^-1, respectively. These results suggest that bioactive compounds from F. oxysporum have the potential as antimicrobial agents, warranting further research.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141854362","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":"Integration of RCA-Based DNA Nanoscaffold with Target Triggered RNA-Cleaving DNAzyme for Sensitive Detection of miRNA21.","authors":"Yuan Li, Xuefei Lv, Hao Jiang, Xiaoqiong Li, Yulin Deng","doi":"10.1007/s12010-024-05022-1","DOIUrl":"https://doi.org/10.1007/s12010-024-05022-1","url":null,"abstract":"<p><p>Cascaded amplification showed promising potential for detection of trace target miRNAs in molecular diagnosis and prevention of many diseases. In this study, miRNA21 was chosen as the target, and rolling circle amplification (RCA)-based DNA nanoscaffold was integrated with target triggered RNA-cleaving DNAzyme for sensitive detection of miRNA21. That is, the H1 probe was bound with the long-chain product of RCA to self-assemble into DNA nanoscaffold. Target miRNA21 triggered the hybridization chain reaction (HCR) located on the nanoscaffold, and led to rapid proximity of DNAzyme fragments modified at both ends of the H2 probe, which realized the cyclic cleavage of self-quenching substrate probe efficiently, and the fluorescence signal was restored. The results demonstrated that the proposed assay was sensitive, 0.76 pM of miRNA21 can be detected. The proposed assay was specific; only one-base mismatched miRNA21 can be effectively recognized, other nucleic acid sequence and the serum matrix did not cause any interference. The proposed assay was accurate; recoveries from 82.1 to 115.0% can be obtained in the spiked fetal bovine serum (FBS). The flexible and programmable characteristics of DNA nanoscaffold and DNAzyme provide a confident and robust strategy for more sensitive nucleic acid detection, and can be developed to be a universal sensing platform for detecting other miRNAs just needing modification on the corresponding sequence of H1 probe in HCR.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141854361","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":"Cyanobacterial Phycocyanin-Based Electrochemical Biosensor for the Detection of the Free Radical Hydrogen Peroxide.","authors":"Sivaprakasam Sivasankari, Mani Vinoth, Abraham David Ravindran","doi":"10.1007/s12010-024-04978-4","DOIUrl":"https://doi.org/10.1007/s12010-024-04978-4","url":null,"abstract":"<p><p>Cyanobacteria are photosynthetic prokaryotes that inhabit extreme environments by modifying their photosensitive chemoreceptors called cyanobacteriochromes (CBCRs) which are linear tetrapyrrole-linked phycobilin molecules. These light-sensitive phycobilin from Spirulina platensis is recognized as a potential photoreceptor tool in optogenetics for monitoring cellular morphogenesis. We prepared and extracted highly fluorescent cyanobacterial phycocyanin (C-PC) by irradiating the culture with ambient red light. The crude phycocyanin was subjected to ion exchange chromatography, and its purity was monitored using UV-visible, fluorescence, and FT-IR spectroscopy methods. In the conventional method, red light-induced C-PC exhibited strong antioxidant activity against H₂O₂, with 88.7% in vitro scavenging activity without requiring any other preservatives. Interestingly, this red light-acclimated phycocyanin was applied as a biosensing material for the detection of the free radical hydrogen peroxide (H₂O₂) using the enzyme horseradish peroxidase (HRP) as a mediator. The modified C-PC-HRP glassy carbon electrode (GCE) can detect H₂O₂ from 0.1 to 1600 µM. The lowest possible detection limit of the electrode for H₂O₂ was 19 nM. This electrode was used to detect free radical H₂O₂ in blood serum samples. The microstructure of the lyophilized PC under SEM showed a flat crystal pattern, which enabled the immobilization of HRP on the electrode surface and electron transfer.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141756468","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":"Kunzea Ericoides (Kanuka) Leaf Extracts Show Moisturisation, Antioxidant, and UV Protection Effects in HaCaT Cells and Anti-melanogenesis Effects in B16F10 Cells.","authors":"Xuefeng He, Peishan Li, Shixin Zhao, Hengdeng Liu, Weijian Tang, Julin Xie, Jinming Tang","doi":"10.1007/s12010-024-04989-1","DOIUrl":"https://doi.org/10.1007/s12010-024-04989-1","url":null,"abstract":"<p><p>Kunzea ericoides (kanuka) products are well-known for their potent medicinal values in antioxidant and anti-inflammatory applications. The present study identified various compounds, such as chlorogenic acid, gallic acid, quercetin, and (E)-ferulic acid in the kanuka leaf extract, showing its potential use in maintaining skin health. The influence of kanuka leaf extract upon epidermal cells concerning cytotoxicity and in vitro activities of moisturisation, antioxidation, UV protection, and anti-melanogenesis effects were explored in the study. Kanuka leaf extract demonstrated significant promotion in the proliferation of HaCaT and B16F10 cells. After incubation with kanuka leaf extract, the content of ROS and DPPH in HaCaT was significantly decreased; at the same time, more SOD was produced. Furthermore, hyaluronidase-1 (HYAL-1) and HYAL-4 expressions were inhibited, while the aquaporin 3 (AQP-3) content was significantly increased in HaCaT. Kanuka leaf extract also inhibited the expressions of matrix metalloproteinases-1 (MMP-1) and MMP-14 in UV-induced HaCaT cells. In the B16F10 cell line, melanin and tyrosinase production were decreased under the presence of kanuka leaf extract, and the expressions of microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TYRP-1), and TYRP-2 were also inhibited. The study validated kanuka leaf extract as an effective natural product against photoaging and melanogenesis.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141618868","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":"Genomic Insights into the Landfill Microbial Community: Denitrifying Activity Supporting One-Carbon Utilization.","authors":"Bhagyashri J Poddar, Anshuman A Khardenavis","doi":"10.1007/s12010-024-04980-w","DOIUrl":"https://doi.org/10.1007/s12010-024-04980-w","url":null,"abstract":"<p><p>In spite of the developments in understanding of denitrifying methylotrophy in the recent years, challenges still exist in unravelling the overall biochemistry of nitrate-dependent methane oxidation in novel or poorly characterized/not-yet-cultured bacteria. In the present study, landfill site was mined for novel C1-carbon-metabolizing bacteria which can use nitrate/nitrite as an electron acceptor. A high-throughput rapid plate assay identified three bacterial isolates with eminent ability for nitrate-dependent methane metabolism under anaerobic conditions. Taxonomic identification by whole-genome sequence-based overall genome relatedness indices accurately assigned the isolates AAK_M13, AAK_M29, and AAK_M39 at the species level to Enterobacter cloacae, Bacillus subtilis, and Bacillus halotolerans, respectively. Several genes encoding sub-components involved in alcohol utilization and denitrification pathways, such as adh, fdh, fdo, nar, nir, and nor, were identified in all the genomes. Though no gene clusters encoding MMO/AMO were annotated, sequencing of PCR amplicons revealed similarity with pMMO/AMO gene using translated nucleotide sequence of strains AAK_M29 and AAK_M39, while strain AAK_M13 showed similarity with XRE family transcriptional regulator. This suggests the horizontal gene transfer and/or presence of a truncated version of a housekeeping enzyme encoded by genes exhibiting partial sequence similarity with pMMO genes that mimicked its function at greenhouse gas emission sites. Owing to lack of conclusive evidence for presence of methane metabolism genes in the selected isolates, further experiment was performed to validate their nitrate-dependent methane oxidation capacities. Bacillus subtilis AAK_M29, Bacillus halotolerans AAK_M39, and Enterobacter cloacae AAK_M13 could oxidize 60%, 75%, and 85% of the added methane respectively accompanied by high nitrate reduction (56-62%) thus supporting the correlation between these two activities. The remarkable ability of these isolates for nitrate-dependent methane metabolism has highlighted their role in ecological contribution and biotechnological potential to serve as methane and nitrate sinks in the landfill sites.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141557717","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":"The Anti-candidal and Absorbtion Performance of PVA/PVP-Based Jania rubens Hydrogel on Candida tropicalis and Some Physicochemical Properties of the Hydrogel.","authors":"Meltem Boran, Elif Erdogan Eliuz, Deniz Ayas","doi":"10.1007/s12010-024-04997-1","DOIUrl":"https://doi.org/10.1007/s12010-024-04997-1","url":null,"abstract":"<p><p>This study was aimed to create a bioactive hydrogel form with PVA/PVP (polyvinyl alcohol/poly(N-vinylpyrrolidone) polymer using acetone and ethanol extractions of Jania rubens red algae and investigate some pharmaceutical properties. The anti-candidal activity and some inhibition performance of J. rubens/PVA/PVP hydrogel were investigated on Candida tropicalis which is one of the important causes of bloodstream infections. The physicochemical properties of J. rubens/PVA/PVP hydrogel were revealed using FTIR and swelling-absorption tests. The volatile compounds of J. rubens extracts were examined by GCMS. By mixing the extracts in equal proportions, PVA/PVP-based hydrogel was prepared. According to the results, Cumulative Drug Release was stable at 25 °C for the first 5 h. The IZ (inhibition zone) and MIC (minimum inhibitory concentration) of J. rubens/PVA/PVP hydrogel were 9.01 mm and 80.20 mg/mL, respectively. It was found that logarithmic reduction and percent reduction were seen as 1.5 CFU/mL and 97.5%, respectively, on C. tropicalis exposed to J. rubens/PVA/PVP hydrogel in the first 5 min of the incubation. After exposure of C. tropicalis to J. rubens/PVA/PVP, the number of viable cells transferred from the gel to water was between 76.1 and 73.1% in high glucose medium, while it was between 92.2 and 80.8% for the PVA/PVP hydrogel under the same conditions. As a result, PVA/PVP hydrogel was made bioactive with J. rubens extracts for the first time in this study, and its potential for use as a functional anticandidal hydrogel on C. tropicalis has been demonstrated.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141496702","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":"In Silico Design of CT26 Polytope and its Surface Display by ClearColi™-Derived Outer Membrane Vesicles as a Cancer Vaccine Candidate Against Colon Carcinoma.","authors":"Elham Sharif, Navid Nezafat, Fatemeh Maghsood Ahmadi, Elham Mohit","doi":"10.1007/s12010-024-04971-x","DOIUrl":"https://doi.org/10.1007/s12010-024-04971-x","url":null,"abstract":"<p><p>Simultaneous targeting of several mutations can be useful in colorectal cancer (CRC) due to its heterogeneity and presence of somatic mutations. As CT26 mutations and expression profiles resemble those of human CRC, we focused on designing a polyepitope vaccine based on CT26 neoepitopes. Due to its low immunogenicity, outer membrane vesicles (rOMV) as an antigen delivery system and adjuvant was applied. Herein, based on previous experimental and our in silico studies four CT26 neoepitopes with the ability to bind MHC-I and MHC-II, TCR, and induce IFN-α production were selected. To increase their immunogenicity, the gp70 and PADRE epitopes were added. The order of the neoepitopes was determined through 3D structure analysis using ProSA, Verify 3D, ERRAT, and Ramachandran servers. The stable peptide-protein docking between the selected epitopes and MHC alleles strengthen our prediction. The CT26 polytope vaccine sequence was fused to the C-terminal of cytolysin A (ClyA) anchor protein and rOMVs were isolated from endotoxin-free ClearColi™ strain. The results of the C-ImmSim server showed that the ClyA-CT26 polytope vaccine could induce T and B cells immunity.The ClyA-CT26 polytope was characterized as a soluble, stable, immunogen, and non-allergen vaccine and optimized for expression in ClearColi™ 24 h after induction with 1 mM IPTG at 25 °C. Western blot analysis confirmed the expression of ClyA-CT26 polytope by ClearColi™ and also on ClearColi™-derived rOMVs. In conclusion, we found that ClearColi™-derived rOMVs with CT26 polytope can deliver CRC neoantigens and induce antitumor immunity, but in vivo immunological studies are needed to confirm vaccine efficacy.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141490364","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}