{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T21:15:47Z","timestamp":1789593347188,"version":"build-2803163510"},"reference-count":37,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,12,30]],"date-time":"2024-12-30T00:00:00Z","timestamp":1735516800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Key R&amp;D Program of China","award":["2023YFD1301603"],"award-info":[{"award-number":["2023YFD1301603"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Peptidoglycan (PGN) is a unique component of prokaryotic cell walls with immune-enhancing capacities. Here, we extracted PGN from Corynebacterium glutamicum, a by-product of amino acid fermentation, using the trichloroacetic acid (TCA) method. SDS-PAGE analysis confirmed the presence of PGN, with a band of approximately 28 kDa. Further analysis was conducted through amino acid analysis, FTIR, and MALDI-TOF\/TOF MS, and the results showed that the chemical structural monomer of PGN is NAG-(\u03b2-1,4-)-NAM-l-Ala-d-Glu-l-Lis-d-Ala. The immune activation effects of PGN were evaluated in a RAW264.7 cell model. Our results showed that PGN could increase the secretion level of NO, ROS, and immune regulatory substances, including TNF-\u03b1 and IL-1\u03b2, and up-regulated the mRNA expression of TNF-\u03b1 and iNOS. In addition, PGN stimulated the expression of ERK2, MyD88, RIP2, and the related receptor NOD1 in the NF-\u03baB and MAPK pathways. Comparative RNA sequencing was conducted to analyze the gene expression profiles in RAW264.7 cells. KEGG analysis indicated that most of the genes were enriched in the NF-\u03baB, MAPK, and TNF signaling pathways. Taken together, these findings suggest that PGN may have immune-activating potential for the development and application of immune adjuvants. Importantly, the application of PGN also provides a new way to utilize amino acid fermentation by-products.<\/jats:p>","DOI":"10.3390\/ijms26010237","type":"journal-article","created":{"date-parts":[[2024,12,31]],"date-time":"2024-12-31T07:51:57Z","timestamp":1735631517000},"page":"237","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Structural Characterization and Immune Activation Capacity of Peptidoglycan from Corynebacterium glutamicum in RAW264.7 Cells"],"prefix":"10.3390","volume":"26","author":[{"given":"Xiaoying","family":"Wang","sequence":"first","affiliation":[{"name":"Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Risk Assessment Laboratory of Animal Product Quality Safety Feed Source Factors of the Ministry of Agriculture and Rural Affairs, Institute of Feed Research of Chinese Academy of Agricultural Sciences, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuzhen","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Risk Assessment Laboratory of Animal Product Quality Safety Feed Source Factors of the Ministry of Agriculture and Rural Affairs, Institute of Feed Research of Chinese Academy of Agricultural Sciences, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aijuan","family":"Zheng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Risk Assessment Laboratory of Animal Product Quality Safety Feed Source Factors of the Ministry of Agriculture and Rural Affairs, Institute of Feed Research of Chinese Academy of Agricultural Sciences, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhimin","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Risk Assessment Laboratory of Animal Product Quality Safety Feed Source Factors of the Ministry of Agriculture and Rural Affairs, Institute of Feed Research of Chinese Academy of Agricultural Sciences, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3041-1746","authenticated-orcid":false,"given":"Jiang","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Animal Husbandry and Veterinary Science, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiheng","family":"Zou","sequence":"additional","affiliation":[{"name":"Institute of Animal Husbandry and Veterinary Science, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9120-0977","authenticated-orcid":false,"given":"Guohua","family":"Liu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Risk Assessment Laboratory of Animal Product Quality Safety Feed Source Factors of the Ministry of Agriculture and Rural Affairs, Institute of Feed Research of Chinese Academy of Agricultural Sciences, Beijing 100081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41589-023-01538-5","article-title":"Complete Biosynthesis of the Potent Vaccine Adjuvant QS-21","volume":"20","author":"Martin","year":"2024","journal-title":"Nat. Chem. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1080\/14760584.2021.1991794","article-title":"SARS-CoV-2 Subunit Vaccine Adjuvants and Their Signaling Pathways","volume":"21","author":"Mekonnen","year":"2022","journal-title":"Expert Rev. Vaccines"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1038\/s41392-023-01557-7","article-title":"Vaccine Adjuvants: Mechanisms and Platforms","volume":"8","author":"Zhao","year":"2023","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Rapaka, R.R., Cross, A.S., and McArthur, M.A. (2021). Using Adjuvants to Drive T Cell Responses for Next-Generation Infectious Disease Vaccines. Vaccines, 9.","DOI":"10.3390\/vaccines9080820"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Xiong, B., Chen, X., Tu, J., Han, Z., Meng, X., and Sun, H. (2023). Actinidia Eriantha Polysaccharide Exerts Adjuvant Activity by Targeting Linc-AAM. Int. J. Biol. Macromol., 252.","DOI":"10.1016\/j.ijbiomac.2023.126440"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1016\/j.ijbiomac.2021.03.180","article-title":"Multi-Walled Carbon Nanotube Polysaccharide Modified Hericium Erinaceus Polysaccharide as an Adjuvant to Extend Immune Responses","volume":"182","author":"Ren","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Zhao, D., Chen, X., Wang, L., Zhang, J., Zhao, Z., Yue, N., Zhu, Y., Fei, W., Li, X., and Tan, L. (2023). Bidirectional and Persistent Immunomodulation of Astragalus Polysaccharide as an Adjuvant of Influenza and Recombinant SARS-CoV-2 Vaccine. Int. J. Biol. Macromol., 234.","DOI":"10.1016\/j.ijbiomac.2023.123635"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"102112","DOI":"10.1016\/j.psj.2022.102112","article-title":"Preparation and Sulfate Modified of Lagenaria Siceraria (Molina) Standl Polysaccharide and Its Immune-Enhancing Adjuvant Activity","volume":"101","author":"Wusiman","year":"2022","journal-title":"Poult. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"He, J., Zhu, T., Mao, N., Cai, G., Gu, P., Song, Z., Lu, X., Yang, Y., and Wang, D. (2024). Cistanche Deserticola Polysaccharide-Functionalized Dendritic Fibrous Nano-Silica as Oral Vaccine Adjuvant Delivery Enhancing Both the Mucosal and Systemic Immunity. Int. J. Biol. Macromol., 262.","DOI":"10.1016\/j.ijbiomac.2024.129982"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Sudo, H., Tokunoh, N., Tsujii, A., Kawashima, S., Hayakawa, Y., Fukushima, H., Takahashi, K., Koshizuka, T., and Inoue, N. (2023). The Adjuvant Effect of Bacterium-like Particles Depends on the Route of Administration. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1082273"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1016\/S0264-410X(02)00547-9","article-title":"Adjuvant Activity of Peptidoglycan Monomer and Its Metabolic Products","volume":"21","author":"Halassy","year":"2003","journal-title":"Vaccine"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1111\/j.1574-6976.2007.00094.x","article-title":"Peptidoglycan Structure and Architecture","volume":"32","author":"Vollmer","year":"2008","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1146\/annurev-micro-020518-120056","article-title":"Growth and Division of the Peptidoglycan Matrix","volume":"75","author":"Rohs","year":"2021","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1128\/MMBR.62.1.181-203.1998","article-title":"Growth of the Stress-Bearing and Shape-Maintaining Murein Sacculus of Escherichia Coli","volume":"62","year":"1998","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1038\/s41586-020-2236-6","article-title":"The Architecture of the Gram-Positive Bacterial Cell Wall","volume":"582","author":"Burns","year":"2020","journal-title":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1038\/s41579-020-0366-3","article-title":"Regulation of Peptidoglycan Synthesis and Remodelling","volume":"18","author":"Egan","year":"2020","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Bastos, P.A.D., Wheeler, R., and Boneca, I.G. (2021). Uptake, Recognition and Responses to Peptidoglycan in the Mammalian Host. FEMS Microbiol. Rev., 45.","DOI":"10.1093\/femsre\/fuaa044"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"108796","DOI":"10.1016\/j.fsi.2023.108796","article-title":"A Detailed Look at the Impacts of Biofloc on Immunological and Hematological Parameters and Improving Resistance to Diseases","volume":"137","author":"Khanjani","year":"2023","journal-title":"Fish Shellfish Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.fsi.2021.06.001","article-title":"Enhanced immunity and hemocytes proliferation by three immunostimulants in tri-spine horseshoe crab Tachypleus tridentatus","volume":"115","author":"Liu","year":"2021","journal-title":"Fish Shellfish Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"440","DOI":"10.3748\/wjg.v7.i3.440","article-title":"Influence of Whole Peptidoglycan of Bifidobacterium on Cytotoxic Effectors Produced by Mouse Peritoneal Macrophages","volume":"7","author":"Wang","year":"2001","journal-title":"World J. Gastroenterol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1016\/j.intimp.2010.04.005","article-title":"Comparative Study of Structurally Related Peptidoglycan Monomer and Muramyl Dipeptide on Humoral IgG Immune Response to Ovalbumin in Mouse","volume":"10","author":"Habjanec","year":"2010","journal-title":"Int. Immunopharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"113622","DOI":"10.1016\/j.envres.2022.113622","article-title":"The Soil Bacterium, Corynebacterium glutamicum, from Biosynthesis of Value-Added Products to Bioremediation: A Master of Many Trades","volume":"213","author":"Ray","year":"2022","journal-title":"Environ. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1038\/ja.2017.142","article-title":"Our Microbes Not Only Produce Antibiotics, They Also Overproduce Amino Acids","volume":"71","author":"Sanchez","year":"2018","journal-title":"J. Antibiot."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1016\/j.carbpol.2013.04.028","article-title":"Structure and Anti-Inflammatory Capacity of Peptidoglycan from Lactobacillus Acidophilus in RAW-264.7 Cells","volume":"96","author":"Wu","year":"2013","journal-title":"Carbohydr. Polym."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.tim.2019.01.009","article-title":"Elucidating Peptidoglycan Structure: An Analytical Toolset","volume":"27","author":"Carlson","year":"2019","journal-title":"Trends Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4227","DOI":"10.1016\/j.febslet.2010.09.009","article-title":"Role of Prostaglandin E2 in Peptidoglycan Mediated iNOS Expression in Mouse Peritoneal Macrophages in Vitro","volume":"584","author":"Dahiya","year":"2010","journal-title":"FEBS Lett."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Song, X., Li, F., Zhang, M., Xia, Y., Ai, L., and Wang, G. (2022). Effect of D-Ala-Ended Peptidoglycan Precursors on the Immune Regulation of Lactobacillus Plantarum Strains. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.825825"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e2004433","DOI":"10.1002\/advs.202004433","article-title":"Cytokines: From Clinical Significance to Quantification","volume":"8","author":"Liu","year":"2021","journal-title":"Adv. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/S0168-1605(98)00047-6","article-title":"Components of Gut Bacteria as Immunomodulators","volume":"41","author":"Hamann","year":"1998","journal-title":"Int. J. Food Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1016\/j.intimp.2015.02.006","article-title":"Non-Viable Immunobiotic Lactobacillus Rhamnosus CRL1505 and Its Peptidoglycan Improve Systemic and Respiratory Innate Immune Response during Recovery of Immunocompromised-Malnourished Mice","volume":"25","author":"Kolling","year":"2015","journal-title":"Int. Immunopharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1007\/s00253-022-11795-4","article-title":"Peptidoglycan-Based Immunomodulation","volume":"106","author":"Sun","year":"2022","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1021\/acscentsci.1c00200","article-title":"Bacterial Peptidoglycan Fragments Differentially Regulate Innate Immune Signaling","volume":"7","author":"Bersch","year":"2021","journal-title":"ACS Cent. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1038\/nri.2017.136","article-title":"Peptidoglycan Recognition by the Innate Immune System","volume":"18","author":"Wolf","year":"2018","journal-title":"Nat. Rev. Immunol."},{"key":"ref_34","first-page":"1300","article-title":"A new morphologically characterized cell wall preparation (whole peptidoglycan) from Bifidobacterium infantis with a higher efficacy on the regression of an established tumor in mice","volume":"45","author":"Sekine","year":"1985","journal-title":"Cancer Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"115332","DOI":"10.1016\/j.carbpol.2019.115332","article-title":"An Improved Phenol-Sulfuric Acid Method for the Determination of Carbohydrates in the Presence of Persulfate","volume":"227","author":"Zhang","year":"2020","journal-title":"Carbohydr. Polym."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"ref_37","first-page":"297","article-title":"On the Morgan-Elson Reaction","volume":"74","author":"Leaback","year":"1963","journal-title":"Biochim. Biophys Acta"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/1\/237\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:56:53Z","timestamp":1760115413000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/1\/237"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,30]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["ijms26010237"],"URL":"https:\/\/doi.org\/10.3390\/ijms26010237","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,30]]}}}