{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T07:06:22Z","timestamp":1790060782698,"version":"4.0.1"},"reference-count":31,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["iubmb.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech and App Biochem"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Streptococcus suis<\/jats:italic>\n                    serotype 2 (\n                    <jats:italic>S. suis<\/jats:italic>\n                    2) is an important zoonotic pathogen, with its capsular polysaccharide (CPS) serving as a key virulence factor. However, the regulatory mechanisms of CPS biosynthesis remain unclear. This study aimed to prepare and characterize the capsular synthesis regulatory factor Cps2D from\n                    <jats:italic>S. suis<\/jats:italic>\n                    2 and investigate its enzymatic activity. Bioinformatics analysis using ProtParam and Alpha Fold revealed Cps2D as a hydrophilic, stable protein of 242 amino acids, with a molecular weight of 28\u00a0kDa and a pI of 8.69. It was annotated as a tyrosine protein phosphatase, primarily composed of \u03b1\u2010helices and random coils, and structurally homologous to the protein model 3qy8.1.A, suggesting potential interactions with glycosyltransferases involved in capsule synthesis. The\n                    <jats:italic>cps2D<\/jats:italic>\n                    gene was amplified from\n                    <jats:italic>S. suis<\/jats:italic>\n                    2 strain 05ZYH33 and cloned into the pET30a vector, and the recombinant protein was successfully purified via Ni\n                    <jats:sup>2+<\/jats:sup>\n                    \u2010NTA affinity chromatography and confirmed by SDS\u2010PAGE and Western blot. Enzymatic characterization using\n                    <jats:italic>p<\/jats:italic>\n                    \u2010nitrophenyl phosphate as a substrate showed that purified Cps2D exhibited phosphatase activity of 81.64 \u00b1 0.72\u00a0U\/L. The enzyme showed maximal activity at pH 10.0 and 37\u00b0C. Among the divalent metal ions tested, Mg\n                    <jats:sup>2<\/jats:sup>\n                    <jats:sup>+<\/jats:sup>\n                    produced the strongest stimulatory effect, with 5\u00a0mM Mg\n                    <jats:sup>2<\/jats:sup>\n                    <jats:sup>+<\/jats:sup>\n                    yielding the highest activity. Kinetic analysis under optimized conditions gave a\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>m<\/jats:sub>\n                    of 1.367\u00a0mM,\n                    <jats:italic>V<\/jats:italic>\n                    <jats:sub>max<\/jats:sub>\n                    of 1.012 \u00d7 10\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>2<\/jats:sup>\n                    \u00a0mM\u00a0min\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>1<\/jats:sup>\n                    ,\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>cat<\/jats:sub>\n                    of 0.4819\u00a0min\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>1<\/jats:sup>\n                    , and\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>cat<\/jats:sub>\n                    \/\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>m<\/jats:sub>\n                    of 0.3525\u00a0min\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>1<\/jats:sup>\n                    \u00a0mM\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>1<\/jats:sup>\n                    . These results establish a soluble prokaryotic expression and purification strategy for Cps2D and provide the first biochemical evidence supporting its phosphatase activity in vitro. The purified recombinant protein will facilitate future structural and mechanistic studies of phosphorylation\u2010dependent regulation of capsule\u2010associated proteins in\n                    <jats:italic>S. suis<\/jats:italic>\n                    serotype 2.\n                  <\/jats:p>","DOI":"10.1002\/bab.70213","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T06:25:12Z","timestamp":1790058312000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Investigation of the Tyrosine Phosphatase Activity of Cps2D From\n                    <i>Streptococcus suis<\/i>\n                    Serotype 2"],"prefix":"10.1002","author":[{"given":"Jiawen","family":"Ping","sequence":"first","affiliation":[{"name":"College of Life and Geographic Sciences, Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region Kashi University  Kashi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingbing","family":"Lu","sequence":"additional","affiliation":[{"name":"College of Life and Geographic Sciences, Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region Kashi University  Kashi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingwen","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Life and Geographic Sciences, Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region Kashi University  Kashi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Life and Geographic Sciences, Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region Kashi University  Kashi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chengliang","family":"Tang","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Hua Dong Research Institute for Medicine and Biotechnics  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yutao","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Life and Geographic Sciences, Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region Kashi University  Kashi China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiuzhen","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Hua Dong Research Institute for Medicine and Biotechnics  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feng","family":"Zheng","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Hua Dong Research Institute for Medicine and Biotechnics  Nanjing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hua","family":"Ni","sequence":"additional","affiliation":[{"name":"College of Life and Geographic Sciences, Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region Kashi University  Kashi China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1005870317757"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens5030045"},{"issue":"6","key":"e_1_2_10_4_1","article-title":"Streptococcus suis, an Important Pig Pathogen and Emerging Zoonotic Agent\u2014An Update on the Worldwide Distribution Based on Serotyping and Sequence Typing","volume":"3","author":"Goyette\u2010desjardins G.","year":"2014","journal-title":"Emerging Microbes & 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and Basic Biological Characteristics of a cps2D Gene Knockout Strain of Streptococcus suis Serotype 2","volume":"38","author":"Liu X. M.","year":"2022","journal-title":"Chinese Journal of Zoonoses"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0000315"},{"issue":"12","key":"e_1_2_10_19_1","first-page":"1283","article-title":"Bioinformatics Analysis and Recombinant Protein Preparation of the Rhamnosyltransferase Cps2F in Streptococcus suis Serotype 2","volume":"37","author":"Lin M.","year":"2024","journal-title":"Journal of Medical Research: Combat Trauma Care"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.26.4.477"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.62.081307.162944"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.GPP3\u20100019\u20102018"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01056\u201009"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00740\u201010"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.101760"},{"key":"e_1_2_10_26_1","unstructured":"M.Lin \u201cStudy on the Biological Functions of Components Cps2B Cps2C and Cps2D in the Tyrosine Kinase\/Phosphatase System of Streptococcus suis Serotype 2 \u201d (Master's thesis Nanjing Normal University 2023)."},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1109201108"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1093\/femsre\/fuaa067"},{"issue":"6","key":"e_1_2_10_29_1","first-page":"835","article-title":"Effects of Metal Ions on Bovine Intestinal Alkaline Phosphatase","volume":"28","author":"Zhang Y.","year":"2009","journal-title":"Sichuan Journal of Animal Sciences"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1586\/14760584.2015.1101349"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0378\u20101135(01)00452\u20107"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0036312"}],"container-title":["Biotechnology and Applied Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bab.70213","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/bab.70213","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bab.70213","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T06:25:15Z","timestamp":1790058315000},"score":1,"resource":{"primary":{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/10.1002\/bab.70213"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,21]]},"references-count":31,"alternative-id":["10.1002\/bab.70213"],"URL":"https:\/\/doi.org\/10.1002\/bab.70213","archive":["Portico"],"relation":{},"ISSN":["0885-4513","1470-8744"],"issn-type":[{"value":"0885-4513","type":"print"},{"value":"1470-8744","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,21]]},"assertion":[{"value":"2025-11-06","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-08","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-21","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"bab.70213"}}