{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T03:41:40Z","timestamp":1788406900754,"version":"build-2803163510"},"reference-count":30,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,3,6]],"date-time":"2025-03-06T00:00:00Z","timestamp":1741219200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Digestive enzymes, such as proteases and lipases, are widely recognized for their crucial roles in the ripening and production of fermented foods. Digestive enzymes are also used as supplements in nonruminant livestock to enhance feed digestion and promote animal growth. However, information on the effects of exogenous digestive enzymes on gut health and disease remains limited. Notably, recent studies show that consuming proteases and lipases can increase the levels of beneficial bacteria and short-chain fatty acids in rodent gut. These findings led us to hypothesize that intestinal proteases and lipases play beneficial roles by enriching beneficial bacteria. To examine this hypothesis, we reviewed recent studies on the potential effects of exogenous digestive enzymes on gut microbiota composition and overall health. Consistent with the hypothesis, all 13 studies in this review reported significant improvements in animal gut microbiota composition with the dietary supplementation of proteases and lipases. Additionally, the possible mechanisms of the prebiotic-like effects of the enzymes through increased nutrient digestion were discussed. This review explores how exogenous proteases and lipases influence gut microbiota composition and overall health. This is the first review to provide insights into the potential roles of exogenous digestive enzymes as prebiotics.<\/jats:p>","DOI":"10.3390\/nu17050924","type":"journal-article","created":{"date-parts":[[2025,3,6]],"date-time":"2025-03-06T09:59:17Z","timestamp":1741255157000},"page":"924","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Potential Roles of Exogenous Proteases and Lipases as Prebiotics"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2279-832X","authenticated-orcid":false,"given":"Yongshou","family":"Yang","sequence":"first","affiliation":[{"name":"School of Life Sciences, Anhui University, Hefei 230036, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6196-5186","authenticated-orcid":false,"given":"Thanutchaporn","family":"Kumrungsee","sequence":"additional","affiliation":[{"name":"Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima 739-8528, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yukako","family":"Okazaki","sequence":"additional","affiliation":[{"name":"Faculty of Human Life Sciences, Fuji Women\u2019s University, Ishikari 061-3204, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshiro","family":"Watanabe","sequence":"additional","affiliation":[{"name":"Faculty of Human Health, Sonoda Women\u2019s University, Amagasaki 661-0012, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junji","family":"Inoue","sequence":"additional","affiliation":[{"name":"Ahjikan Co., Ltd., Shoko Center, Hiroshima 733-0833, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takafumi","family":"Iguchi","sequence":"additional","affiliation":[{"name":"R & D Division Yaegaki Biotechnology, Inc., Himeji 679-4298, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5161-9880","authenticated-orcid":false,"given":"Shinji","family":"Fukuda","sequence":"additional","affiliation":[{"name":"Institute for Advanced Biosciences, Keio University, Tsuruoka 997-0052, Japan"},{"name":"Transborder Medical Research Center, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan"},{"name":"Gut Environmental Design Group, Kanagawa Institute of Industrial Science and Technology, Kawasaki 210-0821, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manabu","family":"Kuroda","sequence":"additional","affiliation":[{"name":"Amano Enzyme Inc., Nagoya 460-8630, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyoichi","family":"Nishio","sequence":"additional","affiliation":[{"name":"Amano Enzyme Inc., Nagoya 460-8630, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shotaro","family":"Yamaguchi","sequence":"additional","affiliation":[{"name":"Amano Enzyme Inc., Nagoya 460-8630, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5467-3126","authenticated-orcid":false,"given":"Norihisa","family":"Kato","sequence":"additional","affiliation":[{"name":"Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima 739-8528, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"543","DOI":"10.3136\/fstr.27.543","article-title":"Lipase and protease activities in Koji cheeses surface-ripened with Aspergillus strains","volume":"27","author":"Suzuki","year":"2021","journal-title":"Food Sci. Technol. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.anifeedsci.2016.04.015","article-title":"Toward optimal value creation through the application of exogenous mono-component protease in the diets of non-ruminants","volume":"221","author":"Cowieson","year":"2016","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_3","unstructured":"Denhard, M. (2025, January 06). Digestive Enzymes and Enzyme Supplements. Available online: https:\/\/www.hopkinsmedicine.org\/health\/wellness-and-prevention\/digestive-enzymes-and-digestive-enzyme-supplements."},{"key":"ref_4","first-page":"20","article-title":"Digestive Enzyme Supplements","volume":"26","author":"Evans","year":"2025","journal-title":"Today\u2019s Dietit."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"715","DOI":"10.3892\/br.2015.490","article-title":"Beneficial effects of protease preparations derived from Aspergillus on the colonic luminal environment in rats consuming a high-fat diet","volume":"3","author":"Yang","year":"2015","journal-title":"Biomed. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"110579","DOI":"10.1016\/j.biopha.2020.110579","article-title":"Candida rugosa lipase alters the gastrointestinal environment in wild-type mice","volume":"130","author":"Menden","year":"2020","journal-title":"Biomed. Pharmacother."},{"key":"ref_7","unstructured":"Howell, E. (2014). Food Enzymes for Health and Longevity, Lotus Press, Division of Lotus Brands Inc.. [3rd ed.]. Revised and Enlarged."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"187","DOI":"10.2174\/138920021702160114150137","article-title":"Digestive Enzyme Supplementation in Gastrointestinal Diseases","volume":"17","author":"Ianiro","year":"2016","journal-title":"Curr. Drug Metab."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1007\/s40005-022-00605-8","article-title":"Digestive enzyme supplementation in prescription drugs, over-the-counter drugs, and enzyme foods","volume":"53","author":"Park","year":"2023","journal-title":"J. Pharm. Investig."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.nutres.2017.06.004","article-title":"Consumption of an acid protease derived from Aspergillus oryzae causes bifidogenic effect in rats","volume":"44","author":"Yang","year":"2017","journal-title":"Nutr. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1901","DOI":"10.1080\/09168451.2019.1627183","article-title":"Feeding Aspergillus protease preparation combined with adequate protein diet to rats increases levels of cecum gut-protective amino acids, partially linked to Bifidobacterium and Lactobacillus","volume":"83","author":"Yang","year":"2019","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Yang, Y., Kumrungsee, T., Kato, N., Fukuda, S., Kuroda, M., and Yamaguchi, S. (2021). Supplemental Aspergillus Lipase and Protease Preparations Display Powerful Bifidogenic Effects and Modulate the Gut Microbiota Community of Rats. Fermentation, 7.","DOI":"10.3390\/fermentation7040294"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Kostiuchenko, O., Kravchenko, N., Markus, J., Burleigh, S., Fedkiv, O., Cao, L., Letasiova, S., Skibo, G., H\u00e5llenius, F.F., and Prykhodko, O. (2022). Effects of Proteases from Pineapple and Papaya on Protein Digestive Capacity and Gut Microbiota in Healthy C57BL\/6 Mice and Dose-Manner Response on Mucosal Permeability in Human Reconstructed Intestinal 3D Tissue Model. Metabolites, 12.","DOI":"10.3390\/metabo12111027"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.aninu.2021.12.003","article-title":"Effects of dietary supplementation with essential oils and protease on growth performance, antioxidation, inflammation and intestinal function of weaned pigs","volume":"9","author":"Peng","year":"2022","journal-title":"Animal Nutr."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"101052","DOI":"10.1016\/j.animal.2023.101052","article-title":"Dietary supplementation of proteases on growth performance, nutrient digestibility, blood characteristics and gut microbiota of growing pigs fed sorghum-based diets","volume":"18","author":"Peng","year":"2024","journal-title":"Animal"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s42523-023-00287-z","article-title":"Dietary novel alkaline protease from Bacillus licheniformis improves broiler meat nutritional value and modulates intestinal microbiota and metabolites","volume":"6","author":"Yi","year":"2024","journal-title":"Anim. Microbiome"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Liu, T., Ma, W., Wang, J., Wei, Y., Wang, Y., Luo, Z., Zhang, Y., Zeng, X., Guan, W., and Shao, D. (2024). Dietary Protease Supplementation Improved Growth Performance and Nutrients Digestion via Modulating Intestine Barrier, Immunological Response, and Microbiota Composition in Weaned Piglets. Antioxidants, 13.","DOI":"10.3390\/antiox13070816"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Bevilacqua, A., Campaniello, D., Speranza, B., Racioppo, A., Sinigaglia, M., and Corbo, M.R. (2024). An Update on Prebiotics and on Their Health Effects. Foods, 13.","DOI":"10.3390\/foods13030446"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1331","DOI":"10.3389\/fmicb.2015.01331","article-title":"Exploring Amino Acid Auxotrophy in Bifidobacterium bifidum PRL2010","volume":"6","author":"Ferrario","year":"2015","journal-title":"Front. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"13045","DOI":"10.1080\/10408398.2023.2260467","article-title":"The beneficial potential of protein hydrolysates as prebiotic for probiotics and its biological activity: A review","volume":"64","author":"Gao","year":"2024","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Averina, O.V., Poluektova, E.U., Marsova, M.V., and Danilenko, V.N. (2021). Biomarkers and Utility of the Antioxidant Potential of Probiotic Lactobacilli and Bifidobacteria as Representatives of the Human Gut Microbiota. Biomedicines, 9.","DOI":"10.3390\/biomedicines9101340"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4797","DOI":"10.1038\/s41598-022-08840-7","article-title":"Exogenous lipase administration alters gut microbiota composition and ameliorates Alzheimer\u2019s disease-like pathology in APP\/PS1 mice","volume":"12","author":"Menden","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Yang, Y., Jia, H., Ando, C., Kato, H., Kumrungsee, T., Kato, N., Kimoto, A., Fukuda, S., Kuroda, M., and Nishio, K. (2023). Exogenous Penicillium camemberti Lipase Preparation Exerts Prebiotic-like Effects by Increasing Cecal Bifidobacterium and Lactobacillus Abundance in Rats. Fermentation, 9.","DOI":"10.3390\/fermentation9030227"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Song, Y., and Lee, S.-H. (2024). Recent Treatment Strategies for Acute Pancreatitis. J. Clin. Med., 13.","DOI":"10.3390\/jcm13040978"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Pan, Y., Li, J., Fan, Z., Chen, Y., Huang, X., and Wu, D. (2024). New Insights into Chronic Pancreatitis: Potential Mechanisms Related to Probiotics. Microorganisms, 12.","DOI":"10.3390\/microorganisms12091760"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.bbrc.2017.10.130","article-title":"Supplementation of pancreatic digestive enzymes alters the composition of intestinal microbiota in mice","volume":"495","author":"Nishiyama","year":"2018","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"906522","DOI":"10.3389\/fphys.2022.906522","article-title":"The Positive Effects of Exogenous Pancreatin on Growth Performance, Nutrient Digestion and Absorption, and Intestinal Microbiota in Piglets","volume":"13","author":"Liu","year":"2022","journal-title":"Front. Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e2127456","DOI":"10.1080\/19490976.2022.2127456","article-title":"Bifidobacterium spp. and their metabolite lactate protect against acute pancreatitis via inhibition of pancreatic and systemic inflammatory responses","volume":"14","author":"Li","year":"2022","journal-title":"Gut Microbes"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"K\u00e5rlund, A., Paukkonen, I., G\u00f3mez-Gallego, C., and Kolehmainen, M. (2021). Intestinal Exposure to Food-Derived Protease Inhibitors: Digestion Physiology- and Gut Health-Related Effects. Healthcare, 9.","DOI":"10.3390\/healthcare9081002"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"106560","DOI":"10.1016\/j.micpath.2024.106560","article-title":"The proteolytic activity in inflammatory bowel disease: Insight from gut microbiota","volume":"188","author":"Hou","year":"2024","journal-title":"Microb. Pathog."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/5\/924\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:48:18Z","timestamp":1760028498000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/5\/924"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,6]]},"references-count":30,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["nu17050924"],"URL":"https:\/\/doi.org\/10.3390\/nu17050924","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,6]]}}}