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It is synthesized through two different pathways. Cystathionine \u03b2\u2010synthase and cystathionine \u03b3\u2010lyase participate in the reverse transsulfuration pathway, while serine acetyltransferase and cysteine synthase function in the de novo pathway. Two evolutionarily related pyridoxal 5\u2032\u2010phosphate\u2010dependent enzymes, cystathionine \u03b2\u2010synthase <jats:italic>TtCBS1<\/jats:italic> (TTHERM_00558300) and cysteine synthase <jats:italic>TtCSA1<\/jats:italic> (TTHERM_00239430), were identified from a freshwater protozoan <jats:italic>Tetrahymena<\/jats:italic> <jats:italic>thermophila<\/jats:italic>. TtCbs1 contained the N\u2010terminal heme binding domain, catalytic domain, and C\u2010terminal regulatory domain, whereas TtCsa1 consisted of two \u03b1\/\u03b2 domains. The catalytic core of the two enzymes is similar. <jats:italic>TtCBS1<\/jats:italic> and <jats:italic>TtCSA1<\/jats:italic> showed high expression levels in the vegetative growth stage and decreased during the sexual developmental stage. TtCbs1 and TtCsa1 were localized in the cytoplasm throughout different developmental stages. His\u2010TtCbs1 and His\u2010TtCsa1 were expressed and purified in\u00a0vitro. TtCbs1 catalyzed the canonical reaction with the highest velocity and possessed serine sulfhydrylase activity. TtCsa1 showed cysteine synthase activity with high K<jats:sub>m<\/jats:sub> for O\u2010acetylserine and low K<jats:sub>m<\/jats:sub> for sulfide and also had serine sulfhydrylase activity toward serine. Both TtCbs1 and TtCsa1 catalyzed hydrogen sulfide producing. <jats:italic>TtCBS1<\/jats:italic> knockdown and <jats:italic>TtCSA1<\/jats:italic> knockout mutants affected cysteine and glutathione synthesis. TtCbs1 and TtCsa1 are involved in cysteine synthesis through two different pathways in <jats:italic>T<\/jats:italic>. <jats:italic>thermophila<\/jats:italic>.<\/jats:p>","DOI":"10.1111\/jeu.12834","type":"journal-article","created":{"date-parts":[[2020,11,15]],"date-time":"2020-11-15T17:11:41Z","timestamp":1605460301000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Characterization of Cystathionine \u03b2\u2010Synthase TtCbs1 and Cysteine Synthase TtCsa1 Involved in Cysteine Biosynthesis in <i>Tetrahymena<\/i><i>thermophila<\/i>"],"prefix":"10.1111","volume":"68","author":[{"given":"Hongrui","family":"Lv","sequence":"first","affiliation":[{"name":"School of Life Science Shanxi University Taiyuan 030006 China"},{"name":"Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education Institute of Biotechnology Shanxi University Taiyuan 030006 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Xu","sequence":"additional","affiliation":[{"name":"School of Life Science Shanxi University Taiyuan 030006 China"},{"name":"Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education Institute of Biotechnology Shanxi University Taiyuan 030006 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tao","family":"Bo","sequence":"additional","affiliation":[{"name":"Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education Institute of Biotechnology Shanxi University Taiyuan 030006 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1595-9797","authenticated-orcid":false,"given":"Wei","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education Institute of Biotechnology Shanxi University Taiyuan 030006 China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,2,17]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1994.tb18577.x"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-313X.1997.11020251.x"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1998.2037"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-385967-9.00008-6"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/146.1.135"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nutres.2009.10.005"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.C400481200"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M808026200"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.14273"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.14315"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1042\/BCJ20161062"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0040286"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1313683110"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1414545111"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.abb.2004.04.027"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-291X(75)80297-X"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.abb.2007.11.006"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1107\/S1744309112019124"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj1040627"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2017.12.008"},{"key":"e_1_2_6_22_1","first-page":"286","article-title":"Expression, localization and functional analysis of cystathionine \u03b3\u2010lyase Cgl1 from Tetrahymena \n                  thermophila","volume":"35","author":"Han T.","year":"2019","journal-title":"Chin. 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