{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T00:59:01Z","timestamp":1792803541093,"version":"4.5.0"},"reference-count":59,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2022,12,2]],"date-time":"2022-12-02T00:00:00Z","timestamp":1669939200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"EU Interreg North-West Europe","award":["NWE 639"],"award-info":[{"award-number":["NWE 639"]}],"id":[{"id":"https:\/\/ror.org\/027mm1153","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Erasmus student travel grant","award":["NWE 639"],"award-info":[{"award-number":["NWE 639"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>In vitro ACE-1 inhibitory peptides were characterised previously from a number of microalgal species including Spirulina platensis (peptide IAPG), Chlorella vulgaris (peptides FDL, AFL, VVPPA), Isochrysis galbana (peptide YMGLDLK), Chlorella sorokiniana (peptides IW and LW) and indeed Nannochloropsis oculata (peptides GMNNLTP and LEQ). The isolation of protein from Nannochloropsis oculata using a combination of ammonium salt precipitation and xylanase treatment of resulting biomass combined with molecular weight cut off filtration to produce a permeate and characterisation of bioactive peptides is described. The Angiotensin-1-converting enzyme (ACE-1) IC50 value for the generated permeate fraction was 370 \u00b5g\/mL. Ninety-five peptide sequences within the permeate fraction were determined using mass spectrometry and eight peptides were selected for chemical synthesis based on in silico analysis. Synthesized peptides were novel based on a search of the literature and relevant databases. In silico, simulated gastrointestinal digestion identified further peptides with bioactivities including ACE-1 inhibitory peptides and peptides with antithrombotic and calcium\/calmodulin-dependent kinase II (CAMKII) inhibition. This work highlights the potential of Nannochloropsis oculata biomass as both a protein and bioactive peptide resource, which could be harnessed for use in the development of functional foods and feeds.<\/jats:p>","DOI":"10.3390\/biom12121806","type":"journal-article","created":{"date-parts":[[2022,12,5]],"date-time":"2022-12-05T02:56:26Z","timestamp":1670208986000},"page":"1806","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Identification of Bioactive Peptides from Nannochloropsis oculata Using a Combination of Enzymatic Treatment, in Silico Analysis and Chemical Synthesis"],"prefix":"10.3390","volume":"12","author":[{"given":"Maria","family":"Hayes","sequence":"first","affiliation":[{"name":"Food BioSciences Department, Teagasc Food Research Centre, Ashtown, D15 Dublin, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1571-5781","authenticated-orcid":false,"given":"Leticia","family":"Mora","sequence":"additional","affiliation":[{"name":"Instituto de Agroqu\u00edmica y Tecnolog\u00eda de Alimentos, Burjassot CSIC, 46980 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8014-0809","authenticated-orcid":false,"given":"Simona","family":"Lucakova","sequence":"additional","affiliation":[{"name":"Institute of Chemical Process Fundamentals of the Czech Academy of Sciences, Rozvojova 135\/1, 165 02 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,2]]},"reference":[{"key":"ref_1","unstructured":"Darzins, A., Pienkos, P., and Edye, L. (2010). Current Status and Potential for Algal Biofuels Production. IEA Bioenergy Task 39-CommerCializing Liquid Biofuels, IEA."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1021\/bp070371k","article-title":"Biofuels from Microalgae","volume":"24","author":"Li","year":"2008","journal-title":"Biotechnol. Prog."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/s12934-018-0879-x","article-title":"The promising future of microalgae: Current status, challenges, and optimization of a sustainable and renewable industry for biofuels, feed, and other products","volume":"17","author":"Khan","year":"2018","journal-title":"Microb. Cell Fact."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1186\/s13068-020-01718-8","article-title":"Improved lipid productivity in Nannochloropsis gaditana in nitrogen-replete conditions by selection of pale green mutants","volume":"13","author":"Cecchin","year":"2020","journal-title":"Biotechnol. Biofuels"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/0022-0981(91)90007-J","article-title":"The amino-acid and sugar composition of 16 species of microalgae used in mariculture","volume":"145","author":"Brown","year":"1991","journal-title":"J. Exp. Mar. Biol. Ecol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Verspreet, J., Soetemans, L., Gargan, C., Hayes, M., and Bastiaens, L. (2021). Nutritional Profiling and Preliminary Bioactivity Screening of Five Micro-Algae Strains Cultivated in Northwest Europe. Foods, 10.","DOI":"10.3390\/foods10071516"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"103919","DOI":"10.1016\/j.jff.2020.103919","article-title":"Microalgae of the genus Nannochloropsis: Chemical composition and functional implications for human nutrition","volume":"68","author":"Zanella","year":"2020","journal-title":"J. Funct. Foods"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fmars.2016.00068","article-title":"Bioactivity screening of microalgae for antioxidant, anti-inflammatory, anticancer, anti-diabetes and antibacterial activities","volume":"3","author":"Lauritano","year":"2016","journal-title":"Front. Mar. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1166\/jbmb.2013.1264","article-title":"Angiotensin I-Converting Enzyme (ACE) Inhibitory Peptide Isolated from Biodiesel Byproducts of Marine Microalgae, Nannochloropsis Oculata","volume":"7","author":"Heo","year":"2013","journal-title":"J. Biobased Mater. Bioenergy"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.1007\/s40200-020-00681-3","article-title":"Evaluation of the anti-inflammatory and antioxidant effects of the microalgae Nannochloropsis gaditana in streptozotocin-induced diabetic rats","volume":"19","author":"Nacer","year":"2020","journal-title":"J. Diabetes Metab. Disord."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1515\/bmc-2019-0004","article-title":"Antihyperglycemic and antihyperlipidemic activities of Nannochloropsis oculata microalgae in Streptozotocin-induced diabetic rats","volume":"10","author":"Nasirian","year":"2019","journal-title":"Biomol. Concepts"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2266","DOI":"10.1271\/bbb.67.2266","article-title":"Effects of the Marine Unicellular Alga Nannochloropsis sp. to Reduce the Plasma and Liver Cholesterol Levels in Male Rats Fed on Diets with Cholesterol","volume":"67","author":"Werman","year":"2014","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1097\/00005344-199015061-00016","article-title":"Angiotensin-converting enzyme inhibitors and the kallikrein-kinin system","volume":"15","author":"Shimamoto","year":"1990","journal-title":"J. Cardiovasc. Pharmacol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4485","DOI":"10.1021\/jf900803r","article-title":"Anti-hypertensive Nutraceuticals and Functional Foods","volume":"57","author":"Chen","year":"2009","journal-title":"J. Agric. Food Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e023535","DOI":"10.1161\/JAHA.121.023535","article-title":"Angiotensin-Converting Enzyme Inhibitors, Angiotensin II Receptor Blockers, and Outcomes in Patients Hospitalized for COVID-19","volume":"10","author":"Pan","year":"2021","journal-title":"J. Am. Heart Assoc."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1016\/j.sjbs.2015.06.005","article-title":"Milk derived bioactive peptides and their impact on human health\u2014A review","volume":"23","author":"Mohanty","year":"2016","journal-title":"Saudi J. Biol. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1016\/j.foodres.2017.04.027","article-title":"Effect of cooking and in vitro digestion on the antioxidant activity of dry-cured ham by-products","volume":"97","author":"Gallego","year":"2017","journal-title":"Food Res. Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1021\/jf505019z","article-title":"Cardioprotective Cryptides Derived from Fish and Other Food Sources: Generation, Application, and Future Markets","volume":"63","author":"Mora","year":"2015","journal-title":"J. Agric. Food Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1332","DOI":"10.2174\/1381612827666210125155048","article-title":"Bioactive Peptides in Preventative Healthcare: An Overview of Bioactivities and Suggested Methods to Assess Potential Applications","volume":"27","author":"Hayes","year":"2021","journal-title":"Curr. Pharm. Des."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"112275","DOI":"10.1016\/j.biopha.2021.112275","article-title":"Measuring the oral bioavailability of protein hydrolysates derived from food sources: A critical review of current bioassays","volume":"144","author":"Abeer","year":"2021","journal-title":"Biomed. Pharmacother."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2501","DOI":"10.1002\/jsfa.6982","article-title":"Effect of degree of hydrolysis on the bioavailability of corn gluten meal hydrolysates","volume":"95","author":"Jin","year":"2014","journal-title":"J. Sci. Food Agric."},{"key":"ref_22","first-page":"353","article-title":"Nutritional value of proteins from edible seaweed Palmaria palmata (Dulse)","volume":"10","author":"Fleurence","year":"1990","journal-title":"J. Nutr. Biochem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.peptides.2014.07.005","article-title":"Identification of novel dipeptidyl peptidase-IV and angiotensin-I-converting enzyme inhibitory peptides from meat proteins using in silico analysis","volume":"59","author":"Lafarga","year":"2014","journal-title":"Peptides"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7421","DOI":"10.1021\/jf301361c","article-title":"Isolation and characterisation of bioactive pro-peptides with in vitro renin inhibitory activities from the macroalga Palmaria palmata","volume":"60","author":"Fitzgerald","year":"2012","journal-title":"J. Agric. Food Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"S55","DOI":"10.1016\/j.jbiotec.2017.06.985","article-title":"Production of GH11 xylanase for bakery industry by solid state fermentation","volume":"256","author":"Tekkol","year":"2017","journal-title":"J. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Minkiewicz, P., Iwaniak, A., and Darewicz, M. (2019). BIOPEP-UWM Database of Bioactive Peptides: Current Opportunities. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20235978"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1007\/978-1-4939-3524-6_14","article-title":"Large-Scale and Deep Quantitative Proteome Profiling Using Isobaric Labeling Coupled with Two-Dimensional LC\u2013MS\/MS","volume":"1410","author":"Gritsenko","year":"2016","journal-title":"Methods Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8352","DOI":"10.1021\/jf500983n","article-title":"Potential of a renin inhibitory peptide from the red seaweed Palmaria palmata as a functional food ingredient following confirmation and characterisation of a hypotensive effect in spontaneously hypertensive rats","volume":"62","author":"Fitzgerald","year":"2014","journal-title":"J. Agric. Food Chem."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Willenberg, I., Meschede, A.K., Gueler, F., Jang, M.-S., Shushakova, N., and Schebb, N.H. (2015). Food Polyphenols Fail to Cause a Biologically Relevant Reduction of COX-2 Activity. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0139147"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2260","DOI":"10.1128\/AEM.72.3.2260-2264.2006","article-title":"Casein-Derived Antimicrobial Peptides Generated by Lactobacillus acidophilus DPC6026","volume":"72","author":"Hayes","year":"2006","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1007\/BF00003945","article-title":"Comparison of different extractive procedures for proteins from the edible seaweeds Ulva rigida and Ulva rotundata","volume":"7","author":"Fleurence","year":"1995","journal-title":"J. Appl. Phycol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3401","DOI":"10.1007\/s10811-020-02197-y","article-title":"Effects of preservation on protein extraction in four seaweed species","volume":"32","author":"Wijers","year":"2020","journal-title":"J. Appl. Phycol."},{"key":"ref_33","unstructured":"Association of Official Analytical Chemists (1998). Official Methods of Analysis of AOAC International, AOAC International."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1166\/jbmb.2013.1264","article-title":"Angiotensin 1 converting enzyme inhibitory peptides isolated from Biodiesel By Product of Marine Microalga Nannochloropsis oculata","volume":"7","author":"Qian","year":"2013","journal-title":"J. Biobased Mater. BioEnergy"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.1007\/s10811-013-9994-6","article-title":"Purification and identification of novel angiotensin-I converting enzyme (ACE) inhibitory peptides from cultured marine microalgae (Nannochloropsis oculata) protein hydrolysate","volume":"25","author":"Samarakoon","year":"2013","journal-title":"J. Appl. Phycol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"277","DOI":"10.4490\/algae.2016.31.6.29","article-title":"Antiinflammatory and anticancer activities of sterol rich fractions of cultured marine microalga Nannochloropsis oculata","volume":"31","author":"Sanjeewa","year":"2016","journal-title":"Algae"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"012064","DOI":"10.1088\/1755-1315\/404\/1\/012064","article-title":"Identification of bioactive compounds from Nannochloropsis sp.","volume":"404","author":"Fithriani","year":"2020","journal-title":"IOP Conf. Ser. Earth Environ. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Purcell, D., Packer, M.A., and Hayes, M. (2022). Angiotensin-I-Converting Enzyme Inhibitory Activity of Protein Hydrolysates Generated from the Macroalga Laminaria digitata (Hudson) JV Lamouroux 1813. Foods, 11.","DOI":"10.3390\/foods11121792"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Mooney, C., Haslam, N., Pollastri, G., and Shields, D.C. (2012). Towards the Improved Discovery and Design of Functional Peptides: Common Features of Diverse Classes Permit Generalized Prediction of Bioactivity. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0045012"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1922","DOI":"10.3389\/fphar.2021.695401","article-title":"CaMKII Inhibition is a Novel Therapeutic Strategy to Prevent Diabetic Cardiomyopathy","volume":"12","author":"Veitch","year":"2021","journal-title":"Front. Pharmacol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1016\/j.peptides.2012.05.013","article-title":"Preparation and evaluation of antioxidant peptides from ethanol-soluble proteins hydrolysate of Sphyrna lewini muscle","volume":"36","author":"Wang","year":"2012","journal-title":"Peptides"},{"key":"ref_42","first-page":"H188","article-title":"Impact of Commercial Precooking of Common Bean (Phaseolus vulgaris) on the Generation of Peptides, After Pepsin-Pancreatin Hydrolysis, Capable to Inhibit Dipeptidyl Peptidase-IV","volume":"80","author":"Mojica","year":"2014","journal-title":"J. Food Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1038\/nbt0395-265","article-title":"Phage Libraries Displaying Cyclic Peptides with Different Ring Sizes: Ligand Specificities of the RGD-Directed Integrins","volume":"13","author":"Koivunen","year":"1995","journal-title":"Nat. Biotechnol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/S0014-5793(98)00405-0","article-title":"Critical amino acid residues of AIP, a highly specific inhibitory peptide of calmodulin-dependent protein kinase II","volume":"427","author":"Ishida","year":"1998","journal-title":"FEBS Lett."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3565","DOI":"10.1007\/s10811-018-1481-7","article-title":"Production of protein extracts from Swedish red, green, and brown seaweeds, Porphyra umbilicalis K\u00fctzing, Ulva lactuca Linnaeus, and Saccharina latissima (Linnaeus) J. V. Lamouroux using three different methods","volume":"30","author":"Harrysson","year":"2018","journal-title":"J. Appl. Phycol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Echave, J., Fraga-Corral, M., Garcia-Perez, P., Popovi\u0107-Djordjevi\u0107, J., Avdovi\u0107, E.H., Radulovi\u0107, M., Xiao, J., A. Prieto, M., and Simal-Gandara, J. (2021). Seaweed Protein Hydrolysates and Bioactive Peptides: Extraction, Purification, and Applications. Mar. Drugs, 19.","DOI":"10.3390\/md19090500"},{"key":"ref_47","unstructured":"M\u00e6hre, H.K. (2016). Seaweed Proteins\u2014How to Get to Them? Effects of Processing on Nutritional Value, Bioaccessibility and Extractability. [Ph.D. Thesis, The Artic University of Norway (UIT)]. Available online: https:\/\/hdl.handle.net\/10037\/9130."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1007\/s10811-016-0972-7","article-title":"A comparison of protocols for isolating and concentrating protein from the green seaweed Ulva ohnoi","volume":"29","author":"Angell","year":"2017","journal-title":"J. Appl. Phycol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"e20201636","DOI":"10.1590\/0001-3765202220201636","article-title":"Algae as a source of peptides inhibitors of the angiotensin-converting enzyme: A systematic review","volume":"94","author":"Porto","year":"2022","journal-title":"Ann. Braz. Acad. Sci."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3895","DOI":"10.1021\/ja211836t","article-title":"Mechanistic insights into the 1,3 xylanases: Useful enzymes for manipulation of algal biomass","volume":"134","author":"Sakaguchi","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Kroger, M., Klemm, M., and Nelles, M. (2018). Hydrothermal disintegration and extraction of different microalgal species. Energies, 11.","DOI":"10.3390\/en11020450"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1186\/1754-6834-4-36","article-title":"The enhancement of enzymatic hydrolysis of lignocellulosic substrates by the addition of accessory enzymes such as xylanase: Is it an additive or synergistic effect?","volume":"4","author":"Hu","year":"2011","journal-title":"Biotechnol. Biofuels"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Dahiya, S., and Singh, B. (2019). Microbial Xylanases in Bread Making. Reference Module in Food Science, Academic Press.","DOI":"10.1016\/B978-0-08-100596-5.21644-2"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Daliri, E.B.-M., Oh, D.H., and Lee, B.H. (2017). Bioactive peptides. Foods, 6.","DOI":"10.3390\/foods6050032"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Daskaya-Dikmen, C., Yucetepe, A., Karbancioglu-Guler, F., Daskaya, H., and Ozcelik, B. (2017). Angiotensin-I-Converting Enzyme (ACE)-Inhibitory Peptides from Plants. Nutrients, 9.","DOI":"10.3390\/nu9040316"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"5206","DOI":"10.1038\/s41598-021-84820-7","article-title":"Antioxidant properties and inhibition of angiotensin-converting enzyme by highly active peptides from wheat gluten","volume":"11","author":"Liu","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Cunha, S.A., Coscueta, E.R., Nova, P., Silva, J.L., and Pintado, M.M. (2022). Bioactive Hydrolysates from Chlorella vulgaris: Optimal Process and Bioactive Properties. Molecules, 27.","DOI":"10.3390\/molecules27082505"},{"key":"ref_58","first-page":"21","article-title":"CaMKII inhibitors: From research tools to therapeutic agents","volume":"5","author":"Epellicena","year":"2014","journal-title":"Front. Pharmacol."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"D\u00edaz-Roa, A., Espinoza-Culup\u00fa, A., Torres-Garc\u00eda, O., Borges, M.M., Avino, I.N., Alves, F.L., Miranda, A., Patarroyo, M.A., da Silva, P.I., and Bello, F. (2019). Sarconesin II, a New Antimicrobial Peptide Isolated from Sarconesiopsis magellanica Excretions and Secretions. Molecules, 24.","DOI":"10.3390\/molecules24112077"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/12\/1806\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:33:03Z","timestamp":1760146383000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/12\/12\/1806"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,2]]},"references-count":59,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["biom12121806"],"URL":"https:\/\/doi.org\/10.3390\/biom12121806","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,2]]}}}