{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T15:13:16Z","timestamp":1785510796215,"version":"3.56.0"},"reference-count":59,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2023,11,29]],"date-time":"2023-11-29T00:00:00Z","timestamp":1701216000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>Muscle atrophy is a complex physiological condition caused by a variety of reasons, including muscle disuse, aging, malnutrition, chronic diseases, immobilization, and hormonal imbalance. Beyond its effect on physical appearance, this condition significantly reduces the quality of human life, thus warranting the development of preventive strategies. Although exercising is effective in managing this condition, it is applicable only for individuals who can engage in physical activities and are not bedridden. A combination of exercise and nutritional supplementation has emerged as a more advantageous approach. Here, we evaluated the effects of enzyme-assisted hydrolysates of Mytilus edulis prepared using Protamex (PMH), Alcalase (AMH), or Flavourzyme (FMH) in protecting against muscle atrophy in a dexamethasone (Dex)-induced muscular atrophy model in vitro and in vitro. Alcalase-assisted M. edulis hydrolysate (AMH) was the most efficient among the tested treatments and resulted in higher protein recovery (57.06 \u00b1 0.42%) and abundant amino acid composition (43,158 mg\/100 g; 43.16%). AMH treatment also escalated the proliferation of C2C12 cells while increasing the total number of nuclei, myotube coverage, and myotube diameter. These results were corroborated by a successful reduction in the levels of proteins responsible for muscle atrophy, including E3 ubiquitin ligases, and an increase in the expression of proteins associated with muscle hypertrophy, including myogenin and MyHC. These results were further solidified by the successful enhancement of locomotor ability and body weight in zebrafish following AMH treatment. Thus, these findings highlight the potential of AMH in recovery from muscle atrophy.<\/jats:p>","DOI":"10.3390\/md21120623","type":"journal-article","created":{"date-parts":[[2023,11,29]],"date-time":"2023-11-29T12:01:00Z","timestamp":1701259260000},"page":"623","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Alcalase-Assisted Mytilus edulis Hydrolysate: A Nutritional Approach for Recovery from Muscle Atrophy"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-3896-061X","authenticated-orcid":false,"given":"R. P. G. S. K.","family":"Amarasiri","sequence":"first","affiliation":[{"name":"Department of Marine Life Science, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jimin","family":"Hyun","sequence":"additional","affiliation":[{"name":"Department of Marine Life Science, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sang-Woon","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Marine Life Science, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Seafood and Aquatic Science, Gyeongsang National University, Tongyeong 53064, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3299-7266","authenticated-orcid":false,"given":"You-Jin","family":"Jeon","sequence":"additional","affiliation":[{"name":"Department of Marine Life Science, Jeju National University, Jeju 63243, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1180-9094","authenticated-orcid":false,"given":"Jung-Suck","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Seafood and Aquatic Science, Gyeongsang National University, Tongyeong 53064, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1007\/s00223-014-9915-y","article-title":"Skeletal Muscle: A Brief Review of Structure and Function","volume":"96","author":"Frontera","year":"2015","journal-title":"Calcif. Tissue Int."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1016\/j.jamda.2016.04.019","article-title":"Skeletal Muscle Regulates Metabolism via Interorgan Crosstalk: Roles in Health and Disease","volume":"17","author":"Campos","year":"2016","journal-title":"J. Am. Med. Dir. Assoc."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1084","DOI":"10.1016\/j.biocel.2004.10.003","article-title":"Molecular mechanisms involved in muscle wasting in cancer and ageing: Cachexia versus sarcopenia","volume":"37","author":"Busquets","year":"2005","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_4","first-page":"E469","article-title":"Skeletal muscle atrophy and the E3 ubiquitin ligases MuRF1 and MAFbx\/atrogin-1","volume":"307","author":"Bodine","year":"2014","journal-title":"Am. J. Physiol. Metab."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1007\/s13539-013-0105-y","article-title":"Cachexia as a major public health problem: Frequent, costly, and deadly","volume":"4","author":"Farkas","year":"2013","journal-title":"J. Cachex-Sarcopenia Muscle"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"101200","DOI":"10.1016\/j.arr.2020.101200","article-title":"Sarcopenia\u2014Molecular mechanisms and open questions","volume":"65","author":"Wiedmer","year":"2021","journal-title":"Ageing Res. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1111\/j.1749-6632.2010.05809.x","article-title":"Recent advances in the biology and therapy of muscle wasting","volume":"1211","author":"Glass","year":"2010","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1123\/ijsnem.2018-0020","article-title":"IOC Consensus Statement: Dietary Supplements and the High-Performance Athlete","volume":"28","author":"Maughan","year":"2018","journal-title":"Int. J. Sport Nutr. Exerc. Metab."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"G\u00f6ko\u011flu, N. (2021). Shellfish Processing and Preservation, Springer International Publishing.","DOI":"10.1007\/978-3-030-60303-8"},{"key":"ref_10","first-page":"2014","article-title":"The European market for mussels","volume":"115","author":"Monfort","year":"2014","journal-title":"Globefish Res. Programme"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1111\/1541-4337.12312","article-title":"Shellfish: Nutritive value, health benefits, and consumer safety","volume":"16","author":"Venugopal","year":"2017","journal-title":"Compr. Rev. Food Sci. Food Saf."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Hayes, M. (2012). Marine Bioactive Compounds: Sources, Characterization and Applications, Springer.","DOI":"10.1007\/978-1-4614-1247-2"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1111\/jfbc.12070","article-title":"Antioxidant and Anti-Inflammatory Activities of Protein Hydrolysates from Mytilus Edulis and Ultrafiltration Membrane Fractions","volume":"38","author":"Park","year":"2014","journal-title":"J. Food Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1016\/j.foodres.2004.10.002","article-title":"Purification of a radical scavenging peptide from fermented mussel sauce and its antioxidant properties","volume":"38","author":"Rajapakse","year":"2005","journal-title":"Food Res. Int."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"975","DOI":"10.3390\/md11040975","article-title":"Evidence of Anti-Proliferative Activities in Blue Mussel (Mytilus edulis) By-Products","volume":"11","author":"Beaulieu","year":"2013","journal-title":"Mar. Drugs"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1016\/j.fsi.2018.09.009","article-title":"Purification and characterization of an antimicrobial peptide mytichitin-chitin binding domain from the hard-shelled mussel, Mytilus coruscus","volume":"83","author":"Oh","year":"2018","journal-title":"Fish Shellfish. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1016\/j.foodchem.2009.04.077","article-title":"Isolation and characterisation of an anticoagulant oligopeptide from blue mussel, Mytilus edulis","volume":"117","author":"Jung","year":"2009","journal-title":"Food Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1108\/00346650310499730","article-title":"Opportunities and constraints in the functional food market","volume":"33","author":"Gray","year":"2003","journal-title":"Nutr. Food Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/s10067-005-0001-8","article-title":"Systematic review of a marine nutriceutical supplement in clinical trials for arthritis: The effectiveness of the New Zealand green-lipped mussel Perna canaliculus","volume":"25","author":"Cobb","year":"2006","journal-title":"Clin. Rheumatol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"102894","DOI":"10.1016\/j.algal.2022.102894","article-title":"The potent antioxidant effect of Neutrase-assisted hydrolysate from heat-resistant Pyropia yezoensis by molecular weight change","volume":"69","author":"Hyun","year":"2023","journal-title":"Algal Res."},{"key":"ref_21","first-page":"E381","article-title":"A high proportion of leucine is required for optimal stimulation of the rate of muscle protein synthesis by essential amino acids in the elderly","volume":"291","author":"Katsanos","year":"2006","journal-title":"Am. J. Physiol. Metab."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/s40279-014-0242-2","article-title":"The Effects of Protein Supplements on Muscle Mass, Strength, and Aerobic and Anaerobic Power in Healthy Adults: A Systematic Review","volume":"45","author":"Pasiakos","year":"2015","journal-title":"Sports Med."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Ryu, B., Je, J.-G., Jeon, Y.-J., and Yang, H.-W. (2021). Zebrafish Model for Studying Dexamethasone-Induced Muscle Atrophy and Preventive Effect of Maca (Lepidium meyenii). Cells, 10.","DOI":"10.3390\/cells10112879"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Yi, D., Yoshikawa, M., Sugimoto, T., Tomoo, K., Okada, Y., and Hashimoto, T. (2022). Effects of Maca on Muscle Hypertrophy in C2C12 Skeletal Muscle Cells. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23126825"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/0141-0229(95)00232-4","article-title":"Characterization of casein phosphopeptides prepared using alcalase: Determination of enzyme specificity","volume":"19","author":"Nicholas","year":"1996","journal-title":"Enzym. Microb. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1016\/j.foodhyd.2016.05.040","article-title":"Modification of interactions between selected volatile flavour compounds and salt-extracted pea protein isolates using chemical and enzymatic approaches","volume":"61","author":"Kun","year":"2016","journal-title":"Food Hydrocoll."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"C615","DOI":"10.1111\/j.1365-2621.2004.tb09909.x","article-title":"Influence of Hydrolysis Degree on the Functional Properties of Salmon Byproducts Hydrolysates","volume":"69","author":"Gbogouri","year":"2004","journal-title":"J. Food Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1028","DOI":"10.1016\/j.idairyj.2008.05.002","article-title":"Effects of pH, temperature and enzyme-to-substrate ratio on the antigenicity of whey protein hydrolysates prepared by Alcalase","volume":"18","author":"Zheng","year":"2008","journal-title":"Int. Dairy J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2278","DOI":"10.1016\/j.ijbiomac.2018.07.114","article-title":"Alcalase assisted production of novel high alpha-chain gelatin and the functional stability of its hydrogel as influenced by thermal treatment","volume":"118","author":"Zhang","year":"2018","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.procbio.2020.06.019","article-title":"Modifying alcalase activity and stability by immobilization onto chitosan aiming at the production of bioactive peptides by hydrolysis of tilapia skin gelatin","volume":"97","author":"Kimberle","year":"2020","journal-title":"Process. Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1002\/jsfa.7712","article-title":"Enzymatic characterisation of the immobilised Alcalase to hydrolyse egg white protein for potential allergenicity reduction","volume":"97","author":"Yang","year":"2017","journal-title":"J. Sci. Food Agric."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1002\/btpr.2245","article-title":"Enzymatic generation of peptides from potato proteins by selected proteases and characterization of their structural properties","volume":"32","author":"Waglay","year":"2016","journal-title":"Biotechnol. Prog."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.foodchem.2014.12.062","article-title":"Seasonal and geographical variations in the biochemical composition of the blue mussel (Mytilus edulis L.) from Ireland","volume":"177","author":"Grienke","year":"2015","journal-title":"Food Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/0022-0981(75)90066-0","article-title":"Seasonal changes in flesh weight and biochemical composition of mussels (Mytilus edulis L.) in the Conwy Estuary, North Wales","volume":"18","author":"Dare","year":"1975","journal-title":"J. Exp. Mar. Biol. Ecol."},{"key":"ref_35","first-page":"1391","article-title":"Effect of environmental factors on biochemical composition and condition index in the Medieterranean Mussel (Mytilus gallaprovincialis Lamarck, 1819) in the Sinop Region","volume":"27","author":"Kaya","year":"2003","journal-title":"Turk. J. Vet. Anim. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Di Pasquale, M.G. (2007). Amino Acids and Proteins for the Athlete: The Anabolic Edge, CRC Press.","DOI":"10.1201\/9781420043815"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Hall, P.A. (2013). Social Neuroscience and Public Health: Foundations for the Science of Chronic Disease Prevention, Springer.","DOI":"10.1007\/978-1-4614-6852-3"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1249\/JSR.0b013e31817fc0fd","article-title":"Recovery Nutrition: Timing and Composition after Endurance Exercise","volume":"7","author":"Childers","year":"2008","journal-title":"Curr. Sports Med. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1249\/JSR.0000000000000180","article-title":"Nutrition to Support Recovery from Endurance Exercise: Optimal Carbohydrate and Protein Replacement","volume":"14","author":"Moore","year":"2015","journal-title":"Curr. Sports Med. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"S29","DOI":"10.1080\/02640414.2011.619204","article-title":"Dietary protein for athletes: From requirements to optimum adaptation","volume":"29","author":"Phillips","year":"2011","journal-title":"J. Sports Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"111070","DOI":"10.1016\/j.msec.2020.111070","article-title":"3D myotube guidance on hierarchically organized anisotropic and conductive fibers for skeletal muscle tissue engineering","volume":"116","author":"Zhang","year":"2020","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"152873","DOI":"10.1016\/j.phymed.2019.152873","article-title":"4-hydroxy-3-methoxy cinnamic acid accelerate myoblasts differentiation on C2C12 mouse skeletal muscle cells via AKT and ERK 1\/2 activation","volume":"60","author":"Kuppusamy","year":"2019","journal-title":"Phytomedicine"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1080\/10498850.2019.1575940","article-title":"Enzymatic Hydrolysates of Hippocampus abdominalis Regulates the Skeletal Muscle Growth in C2C12 Cells and Zebrafish Model","volume":"28","author":"Kim","year":"2019","journal-title":"J. Aquat. Food Prod. Technol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2240","DOI":"10.1016\/j.procbio.2012.08.019","article-title":"Angiotensin I converting enzyme (ACE) inhibitory peptides from salmon byproduct protein hydrolysate by Alcalase hydrolysis","volume":"47","author":"Ahn","year":"2012","journal-title":"Process. Biochem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1007\/s11694-019-00296-0","article-title":"Effect of enzymatic hydrolysis using endo- and exo-proteases on secondary structure, functional, and antioxidant properties of chickpea protein hydrolysates","volume":"14","author":"Xu","year":"2020","journal-title":"J. Food Meas. Charact."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2143","DOI":"10.1016\/j.ijbiomac.2020.10.060","article-title":"Use of Alcalase in the production of bioactive peptides: A review","volume":"165","author":"Siar","year":"2020","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.freeradbiomed.2015.12.031","article-title":"The role of E3 ubiquitin-ligases MuRF-1 and MAFbx in loss of skeletal muscle mass","volume":"98","author":"Rom","year":"2016","journal-title":"Free. Radic. Biol. Med."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"108893","DOI":"10.1016\/j.cbi.2019.108893","article-title":"Promotion of myotube differentiation and attenuation of muscle atrophy in murine C2C12 myoblast cells treated with teaghrelin","volume":"315","author":"Hsieh","year":"2020","journal-title":"Chem. Interactions"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1002\/ajpa.1330470204","article-title":"Body weight: Its relation to tissue composition, segment distribution, and motor function. I. Interspecific comparisons","volume":"47","author":"Grand","year":"1977","journal-title":"Am. J. Phys. Anthropol."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Hyun, J., Kang, S.-I., Lee, S.-W., Amarasiri, R.P.G.S.K., Nagahawatta, D.P., Roh, Y., Wang, L., Ryu, B., and Jeon, Y.-J. (2023). Exploring the Potential of Olive Flounder Processing By-Products as a Source of Functional Ingredients for Muscle Enhancement. Antioxidants, 12.","DOI":"10.3390\/antiox12091755"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"894","DOI":"10.1210\/jc.2009-1822","article-title":"The Effect of Branched Chain Amino Acids on Skeletal Muscle Mitochondrial Function in Young and Elderly Adults","volume":"95","author":"Tatpati","year":"2010","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_52","unstructured":"Pasin, G., and Miller, S. (2000). Sports Nutrition, U.S. Dairy Export Counci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1152\/physiol.00024.2006","article-title":"The mTOR Pathway in the Control of Protein Synthesis","volume":"21","author":"Wang","year":"2006","journal-title":"Physiology"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1038\/ncb1101-1009","article-title":"Mediation of IGF-1-induced skeletal myotube hypertrophy by PI(3)K\/Akt\/mTOR and PI(3)K\/Akt\/GSK3 pathways","volume":"3","author":"Rommel","year":"2001","journal-title":"Nat. Cell Biol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1111\/ijfs.13323","article-title":"Digestion and bioavailability of bioactive phytochemicals","volume":"52","author":"Jakubczyk","year":"2017","journal-title":"Int. J. Food Sci. Technol."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Kumar, P., Nagarajan, A., and Uchil, P.D. (2018). Analysis of Cell Viability by the MTT Assay. Cold Spring Harb. Protoc., 2018.","DOI":"10.1101\/pdb.prot095505"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1399","DOI":"10.1038\/nprot.2006.224","article-title":"BrdU assay for neurogenesis in rodents","volume":"1","author":"Wojtowicz","year":"2006","journal-title":"Nat. Protoc."},{"key":"ref_58","first-page":"1","article-title":"The Myotube Analyzer: How to assess myogenic features in muscle stem cells","volume":"12","author":"Corvelyn","year":"2022","journal-title":"Skelet. Muscle"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"11.9.1","DOI":"10.1002\/0471140864.ps1109s58","article-title":"Amino acid analysis","volume":"58","author":"Rutherfurd","year":"2009","journal-title":"Curr. Protoc. Protein Sci."}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/21\/12\/623\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:33:58Z","timestamp":1760132038000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/21\/12\/623"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,29]]},"references-count":59,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["md21120623"],"URL":"https:\/\/doi.org\/10.3390\/md21120623","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,29]]}}}