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COX. The ChOx was purified by salting\u2010out and ion\u2010exchange chromatography up to 10.4\u2010fold, with a specific activity of 15\u2009U\/mg with a molecular mass of 59\u2009kDa. The purified ChOx exhibited pH 8.0 and temperature 40\u00b0C for its optimum activity. The enzyme showed stability over a wide pH range and was most stable at pH value 7.0, and at pH 8.0, it retained almost 86% of its initial activity after 3\u2009h of incubation at 37\u00b0C. The enzyme possessed a half\u2010life of 8\u2009h at 37\u00b0C, 7\u2009h at 40\u00b0C, and 3\u2009h at 50\u00b0C. A Lineweaver\u2013Burk plot was calibrated to determine its <jats:italic>K<\/jats:italic><jats:sub>m<\/jats:sub> (0.16\u2009mM) and <jats:italic>V<\/jats:italic><jats:sub>max<\/jats:sub> (18.7\u2009\u03bcmol\u00b7mg<jats:sup>\u22121<\/jats:sup>\u00b7min<jats:sup>\u22121<\/jats:sup>). The ChOx activity was enhanced with Ca<jats:sup>2+<\/jats:sup>, Mg<jats:sup>2+<\/jats:sup>, and Mn<jats:sup>2+<\/jats:sup> while it was inhibited by Hg<jats:sup>2+<\/jats:sup>, Ba<jats:sup>2+<\/jats:sup>, Fe<jats:sup>2+<\/jats:sup>, Cu<jats:sup>2+<\/jats:sup>, and Zn<jats:sup>2+<\/jats:sup> ions. Organic solvents like acetone, <jats:italic>n<\/jats:italic>\u2010butanol, toluene, dimethyl sulfoxide, chloroform, benzene, and methanol were well tolerated by the enzyme while <jats:italic>iso<\/jats:italic>\u2010propanol and ethanol were found to enhance the activity of purified ChOx. ChOx induced cytotoxicity with an IC<jats:sub>50<\/jats:sub> value of 1.78 and 1.88\u2009U\/ml against human RD and U87MG established cell lines, respectively, while broadly sparing the normal human cells.<\/jats:p>","DOI":"10.1002\/jobm.201900365","type":"journal-article","created":{"date-parts":[[2019,11,21]],"date-time":"2019-11-21T13:59:40Z","timestamp":1574344780000},"page":"253-267","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Purification, characterization, and biological cytotoxic activity of the extracellular cholesterol oxidase produced by <i>Castellaniella<\/i> sp. 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