{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:21:47Z","timestamp":1785403307950,"version":"3.56.0"},"reference-count":60,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,5,31]],"date-time":"2024-05-31T00:00:00Z","timestamp":1717113600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Fungal Biol."],"abstract":"<jats:p><jats:italic>Candida albicans<\/jats:italic> is the predominant cause of systemic candidiasis, although other non albicans Candida species are progressively becoming more widespread nowadays. <jats:italic>Candida auris<\/jats:italic> has emerged as a deadly multidrug-resistant fungal pathogen, posing a significant threat to global public health. In the absence of effective antifungal therapies, the development of a vaccine against <jats:italic>C. auris<\/jats:italic> infections is imperative. Enolase, a key glycolytic enzyme, has emerged as a promising vaccine candidate due to its immunogenic properties and essential role in fungal virulence. Herein, full-length Enolase gene sequences from <jats:italic>C. albicans<\/jats:italic> and <jats:italic>C. auris<\/jats:italic> were cloned into suitable expression vector and transformed into <jats:italic>Escherichia coli<\/jats:italic> expression hosts. Recombinant Enolase proteins were successfully expressed and purified using affinity chromatography under native conditions, followed by SDS-PAGE characterization and Western blot analysis. CD spectroscopy verified the existence of expressed proteins in soluble native conformation. Preliminary in silico studies verified the immunogenicity of recombinant Enolase proteins isolated from both <jats:italic>C. albicans<\/jats:italic> and <jats:italic>C. auris<\/jats:italic>. Furthermore, bioinformatics analysis revealed conserved B-cell and T-cell epitopes across <jats:italic>C. albicans<\/jats:italic> and <jats:italic>C. auris<\/jats:italic> Enolase proteins, suggesting potential cross-reactivity and broad-spectrum vaccine efficacy. Our findings are anticipated to play a role in advancing therapeutic as well as diagnostic strategies against systemic candidiasis.<\/jats:p>","DOI":"10.3389\/ffunb.2024.1399546","type":"journal-article","created":{"date-parts":[[2024,5,31]],"date-time":"2024-05-31T04:58:45Z","timestamp":1717131525000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Molecular cloning, expression, and purification, along with in silico epitope analysis of recombinant enolase proteins (a potential vaccine candidate) from Candida albicans and Candida auris"],"prefix":"10.3389","volume":"5","author":[{"given":"Manisha","family":"Shukla","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rohit","family":"Singh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pankaj","family":"Chandley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soma","family":"Rohatgi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,5,31]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"104879","DOI":"10.1016\/j.micpath.2021.104879","article-title":"In-silico design of a multivalent epitope-based vaccine against Candida auris","volume":"155","author":"Akhtar","year":"2021","journal-title":"Microb. Pathog."},{"key":"B2","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1186\/s13054-016-1313-6","article-title":"Diagnosis and management of invasive candidiasis in the ICU: an updated approach to an old enemy","volume":"20","author":"Calandra","year":"2016","journal-title":"Crit. Care"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.3791\/64425","article-title":"A soluble tetrazolium-based reduction assay to evaluate the effect of antibodies on candida tropicalis biofilms","volume":"10","author":"Chandley","year":"2022","journal-title":"J.\u00a0Visual. Exper."},{"key":"B4","doi-asserted-by":"publisher","first-page":"583","DOI":"10.1146\/annurev-micro-032521-015858","article-title":"Candida auris genetics and emergence","volume":"77","author":"Chowdhary","year":"2023","journal-title":"Annu. Rev. Microbiol."},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1002\/eji.202250284","article-title":"Enolase 1 of Candida albicans binds human CD4+ T cells and modulates na\u00efve and memory responses","volume":"53","author":"Daud","year":"2023","journal-title":"Eur. J. Immunol."},{"key":"B6","first-page":"82","article-title":"Pymol: An open-source molecular graphics tool","volume":"40","author":"DeLano","year":"2002","journal-title":"CCP4 Newsl Protein Crystallogr."},{"key":"B7","doi-asserted-by":"publisher","first-page":"2278","DOI":"10.1007\/s00894-014-2278-5","article-title":"AllerTOP v.2\u2013a server for in silico prediction of allergens","volume":"20","author":"Dimitrov","year":"2014","journal-title":"J. Mol. Model."},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-8-4","article-title":"VaxiJen: a server for prediction of protective antigens, tumour antigens and subunit vaccines","volume":"8","author":"Doytchinova","year":"2007","journal-title":"BMC Bioinf."},{"key":"B9","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1111\/fml.1993.111.issue-1","article-title":"Cloning and nucleotide sequence analysis of the Candida albicans enolase gene","volume":"111","author":"Franklyn","year":"1993","journal-title":"FEMS Microbiol. Lett."},{"key":"B10","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1016\/j.ijmm.2015.11.005","article-title":"The glycolytic enzyme enolase represents a plasminogen-binding protein on the surface of a wide variety of medically important fungal species","volume":"306","author":"Funk","year":"2016","journal-title":"Int. J. Med. Microbiol."},{"key":"B11","first-page":"449","article-title":"Characterization of sequence and structural features of the Candida krusei enolase","volume":"8","author":"Gandhi","year":"2008","journal-title":"Silico Biol."},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2015.00920","article-title":"Serological response and diagnostic value of recombinant candida cell wall protein enolase, phosphoglycerate kinase, and \u03b2-glucosidase","volume":"6","author":"He","year":"2015","journal-title":"Front. Microbiol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1186\/s13213-022-01682-8","article-title":"Investigation of the location and secretion features of Candida albicans enolase with monoclonal antibodies","volume":"72","author":"He","year":"2022","journal-title":"Ann. Microbiol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"7079356","DOI":"10.1155\/2020\/7079356","article-title":"An immunoinformatics study to predict epitopes in the envelope protein of SARS-CoV-2","volume":"2020","author":"Jakhar","year":"2020","journal-title":"Can. J. Infect. Dis. Med. Microbiol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"W24","DOI":"10.1093\/nar\/gkx346","article-title":"BepiPred-2.0: improving sequence-based B-cell epitope prediction using conformational epitopes","volume":"45","author":"Jespersen","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"B16","doi-asserted-by":"publisher","first-page":"615","DOI":"10.1099\/jmm.0.05060-0","article-title":"Binding of Candida albicans enolase to plasmin(ogen) results in enhanced invasion of human brain microvascular endothelial cells","volume":"52","author":"Jong","year":"2003","journal-title":"J. Med. Microbiol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1722","DOI":"10.3390\/cells9071722","article-title":"Characteristics of Extracellular Vesicles Released by the Pathogenic Yeast-Like Fungi Candida glabrata, Candida parapsilosis and Candida tropicalis","volume":"9","author":"Karkowska-Kuleta","year":"2020","journal-title":"Cells"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.13297","article-title":"Als3-mediated attachment of enolase on the surface of Candida albicans cells regulates their interactions with host proteins","volume":"23","author":"Karkowska-Kuleta","year":"2021","journal-title":"Cell Microbiol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"345","DOI":"10.1007\/s12275-013-2577-z","article-title":"Candida albicans ENO1 null mutants exhibit altered drug susceptibility, hyphal formation, and virulence","volume":"51","author":"Ko","year":"2013","journal-title":"J.\u00a0Microbiol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"891","DOI":"10.1002\/prot.21770","article-title":"Sequence-similar, structure-dissimilar protein pairs in the PDB","volume":"71","author":"Kosloff","year":"2008","journal-title":"Proteins"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1002829","article-title":"Reliable B cell epitope predictions: impacts of method development and improved benchmarking","volume":"8","author":"Kringelum","year":"2012","journal-title":"PloS Comput. Biol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"318","DOI":"10.1128\/CVI.00396-06","article-title":"Evaluation of a novel enzyme-linked immunosorbent assay to detect immunoglobulin G antibody to enolase for serodiagnosis of invasive candidiasis","volume":"14","author":"La\u00edn","year":"2007","journal-title":"Clin. Vaccine Immunol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"804","DOI":"10.4049\/jimmunol.2200318","article-title":"Soluble Enolase 1 of Candida albicans and Aspergillus fumigatus Stimulates Human and Mouse B Cells and Monocytes","volume":"211","author":"Langenhorst","year":"2023","journal-title":"J. Immunol."},{"key":"B24","doi-asserted-by":"publisher","first-page":"283","DOI":"10.1107\/S0021889892009944","article-title":"PROCHECK-a program to check the stereochemical quality of protein structures","volume":"26","author":"Laskowski","year":"1996","journal-title":"J.\u00a0Appl. Cryst."},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21082903","article-title":"Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans","volume":"21","author":"Leu","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B26","doi-asserted-by":"publisher","first-page":"5526","DOI":"10.1016\/j.vaccine.2011.05.030","article-title":"Immunisation with the glycolytic enzyme enolase confers effective protection against Candida albicans infection in mice","volume":"29","author":"Li","year":"2011","journal-title":"Vaccine"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.02085-22","article-title":"Baicalein Acts against Candida albicans by Targeting Eno1 and Inhibiting Glycolysis","volume":"10","author":"Li","year":"2022","journal-title":"Microbiol. Spectr."},{"key":"B28","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1186\/1471-2334-13-253","article-title":"Diagnostic value of immunoglobulin G antibodies against Candida enolase and fructose-bisphosphate aldolase for candidemia","volume":"13","author":"Li","year":"2013","journal-title":"BMC Infect. Dis."},{"key":"B29","doi-asserted-by":"publisher","first-page":"1127","DOI":"10.1093\/mmy\/myab053","article-title":"Opportunistic yeast pathogen Candida spp.: Secreted and membrane-bound virulence factors","volume":"59","author":"Lim","year":"2021","journal-title":"Med. Mycol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"2632","DOI":"10.1128\/jb.175.9.2632-2639.1993","article-title":"Molecular cloning and characterization of the Candida albicans enolase gene","volume":"175","author":"Mason","year":"1993","journal-title":"J. Bacteriol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"1660","DOI":"10.3390\/vaccines10101660","article-title":"Immunoinformatics-based identification of B and T cell epitopes in RNA-dependent RNA polymerase of SARS-cov-2","volume":"10","author":"Mir","year":"2022","journal-title":"Vaccines (Basel)."},{"key":"B32","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1080\/13693780310001644653","article-title":"Immunogenicity and protective effect of recombinant enolase of Candida albicans in a murine model of systemic candidiasis","volume":"42","author":"Montagnoli","year":"2004","journal-title":"Medical Mycology"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.00634-22","article-title":"ClaID: a rapid method of clade-level identification of the multidrug resistant human fungal pathogen candida auris","volume":"10","author":"Narayanan","year":"2022","journal-title":"Microbiol. Spectr."},{"key":"B34","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1016\/j.micpath.2017.04.009","article-title":"Identification of proteins involved in the adhesion of Candida species to different medical devices","volume":"107","author":"N\u00fa\u00f1ez-Beltr\u00e1n","year":"2017","journal-title":"Microb. Pathog."},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003550","article-title":"Emerging and emerged pathogenic Candida species: beyond the Candida albicans paradigm","volume":"9","author":"Papon","year":"2013","journal-title":"PloS Pathog."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/nrdp.2018.26","article-title":"Invasive candidiasis","volume":"4","author":"Pappas","year":"2018","journal-title":"Nat. Rev. Dis. Prim."},{"key":"B37","doi-asserted-by":"publisher","first-page":"1087","DOI":"10.1080\/14787210.2023.2264500","article-title":"Host defense mechanisms against Candida auris","volume":"21","author":"Pechacek","year":"2023","journal-title":"Expert Rev. Anti Infect. Ther."},{"key":"B38","doi-asserted-by":"publisher","first-page":"5165","DOI":"10.1021\/pr500681x","article-title":"Serum antibody signature directed against Candida albicans Hsp90 and enolase detects invasive candidiasis in non-neutropenic patients","volume":"13","author":"Pitarch","year":"2014","journal-title":"J. Proteome Res."},{"key":"B39","doi-asserted-by":"publisher","first-page":"4304","DOI":"10.1074\/jbc.M117.810440","article-title":"The Candida albicans ENO1 gene encodes a transglutaminase involved in growth, cell division, morphogenesis, and osmotic protection","volume":"293","author":"Reyna-Beltr\u00e1n","year":"2018","journal-title":"J. Biol. Chem."},{"key":"B40","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1111\/j.1470-8744.1999.tb00553.x","article-title":"Generation of a highly immunogenic recombinant enolase of the human opportunistic pathogen Candida albicans","volume":"29","author":"Sandini","year":"1999","journal-title":"Biotechnol. Appl. Biochem."},{"key":"B41","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/microorganisms8071046","article-title":"Moonlighting proteins at the candidal cell surface","volume":"8","author":"Satala","year":"2020","journal-title":"Microorganisms"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21217843","article-title":"Structural insights into the interactions of candidal enolase with human vitronectin, fibronectin and plasminogen","volume":"21","author":"Satala","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B43","doi-asserted-by":"publisher","first-page":"697","DOI":"10.3797\/(ISSN)0036-8709","article-title":"Oral Immunization Against Candidiasis Using Lactobacillus casei Displaying Enolase 1 from Candida albicans","volume":"82","author":"Shibasaki","year":"2014","journal-title":"Sci. Pharm."},{"key":"B44","doi-asserted-by":"publisher","first-page":"999","DOI":"10.3390\/jof7120999","article-title":"Expression and purification along with evaluation of serological response and diagnostic potential of recombinant Sap2 protein from C. parapsilosis for use in systemic candidiasis","volume":"7","author":"Shukla","year":"2021","journal-title":"J. Fungi"},{"key":"B45","doi-asserted-by":"publisher","first-page":"1159","DOI":"10.3390\/vaccines9101159","article-title":"The role of B cells and antibodies against candidate vaccine antigens in invasive candidiasis","volume":"9","author":"Shukla","year":"2021","journal-title":"Vaccines"},{"key":"B46","doi-asserted-by":"publisher","first-page":"3491","DOI":"10.1021\/acsomega.1c05975","article-title":"Expression, purification and refolding of Chikungunya virus full length envelop E2 protein along with B-cell and T-cell epitope analysis using immuno-informatics approaches","volume":"7","author":"Shukla","year":"2022","journal-title":"ACS Omega"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00312-20","article-title":"Vaccination with secreted aspartyl proteinase 2 protein from Candida parapsilosis can enhance survival of mice during Candida tropicalis mediated systemic candidiasis","volume":"88","author":"Shukla","year":"2020","journal-title":"Infect. Immun."},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2014.00066","article-title":"Extracellular enolase of Candida albicans is involved in colonization of mammalian intestinal epithelium","volume":"4","author":"Silva","year":"2014","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007460","article-title":"The NDV-3A vaccine protects mice from multidrug resistant Candida auris infection","volume":"15","author":"Singh","year":"2019","journal-title":"PloS Pathog."},{"key":"B50","doi-asserted-by":"publisher","first-page":"1569","DOI":"10.1093\/jac\/dkad139","article-title":"Invasive candidiasis: current clinical challenges and unmet needs in adult populations","volume":"78","author":"Soriano","year":"2023","journal-title":"J. Antimicrobial. Chemother."},{"key":"B51","doi-asserted-by":"publisher","first-page":"e01588","DOI":"10.1128\/JCM.01588-17","article-title":"Candida auris: an emerging fungal pathogen","volume":"56","author":"Spivak","year":"2018","journal-title":"J. Clin. Microbiol."},{"key":"B52","doi-asserted-by":"publisher","first-page":"2405","DOI":"10.1080\/22221751.2022.2125349","article-title":"Confirmation of fifth Candida auris clade by whole genome sequencing","volume":"11","author":"Spruijtenburg","year":"2022","journal-title":"Emerg. Microbes Infect."},{"key":"B53","doi-asserted-by":"publisher","first-page":"390","DOI":"10.1093\/infdis\/170.2.390","article-title":"Humoral and cellular immune responses to enolase after alimentary tract colonization or intravenous immunization with candida albicans","volume":"170","author":"Sundstrom","year":"1994","journal-title":"J. Infect. Dis."},{"key":"B54","doi-asserted-by":"publisher","first-page":"23293435","DOI":"10.1101\/2023.08.01.23293435","article-title":"Discovery of the sixth Candida auris clade in Singapore","author":"Suphavilai","year":"2023","journal-title":"medRxiv"},{"key":"B55","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1111\/j.1469-0691.1996.tb00198.x","article-title":"Anti-enolase antibodies partially protective against systemic candidiasis in mice","volume":"2","author":"Van Deventer","year":"1996","journal-title":"Clin. Microbiol. Infect."},{"key":"B56","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1128\/jcm.32.1.17-23.1994","article-title":"Diagnostic value of anti-Candida enolase antibodies","volume":"32","author":"van Deventer","year":"1994","journal-title":"J. Clin. Microbiol."},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.00897","article-title":"The Elusive Anti-Candida Vaccine: Lessons from the past and opportunities for the future","volume":"9","author":"Wan Tso","year":"2018","journal-title":"Front. Immunol."},{"key":"B58","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3314\/jjmm.48.1","article-title":"Trends in the epidemiology of invasive fungal infections","volume":"48","author":"Warnock","year":"2007","journal-title":"Jpn. J. Med. Mycol."},{"key":"B59","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.cyto.2015.07.025","article-title":"Beyond Candida albicans: mechanisms of immunity to non-albicans Candida species","volume":"76","author":"Whibley","year":"2015","journal-title":"Cytokine"},{"key":"B60","doi-asserted-by":"publisher","first-page":"13526","DOI":"10.1073\/pnas.0803195105","article-title":"Synthetic glycopeptide vaccines combining beta-mannan and peptide epitopes induce protection against candidiasis","volume":"105","author":"Xin","year":"2008","journal-title":"Proc. Natl. Acad. Sci. U S A."}],"container-title":["Frontiers in Fungal Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/ffunb.2024.1399546\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,7,19]],"date-time":"2024-07-19T10:30:20Z","timestamp":1721385020000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/ffunb.2024.1399546\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,31]]},"references-count":60,"alternative-id":["10.3389\/ffunb.2024.1399546"],"URL":"https:\/\/doi.org\/10.3389\/ffunb.2024.1399546","relation":{},"ISSN":["2673-6128"],"issn-type":[{"value":"2673-6128","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,31]]},"article-number":"1399546"}}