title: A factor converting viable but nonculturable Vibrio cholerae to a culturable state in eukaryotic cells is a human catalase.
abstract: In our previous work, we demonstrated that viable but nonculturable (VBNC) Vibrio cholerae O1 and O139 were converted to culturable by coculture with eukaryotic cells. Furthermore, we isolated a factor converting VBNC V. cholerae to culturable (FCVC) from a eukaryotic cell line, HT-29. In this study, we purified FCVC by successive column chromatographies comprising UNO Q-6 anion exchange, Bio-Scale CHT2-1 hydroxyapatite, and Superdex 200 10/300 GL. Homogeneity of the purified FCVC was demonstrated by SDS-PAGE. Nano-LC MS/MS analysis showed that the purified FCVC was a human catalase. An experiment of RNAi knockdown of catalase mRNA from HT-29 cells and treatment of the purified FCVC with a catalase inhibitor, 3-amino-1,2,4-triazole confirmed that the FCVC was a catalase. A possible role of the catalase in converting a VBNC V. cholerae to a culturable state in the human intestine is discussed.
authors.affiliation: Collaborative Research Center of Okayama University for Infectious Diseases in India, Okayama University, Kolkata, India.
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Senoh
authors.affiliation: Research Institute, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Hamabata
authors.affiliation: Collaborative Research Center of Okayama University for Infectious Diseases in India, Okayama University, Kolkata, India.
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Takeda
mesh_terms: D002374:Catalase / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D002853:Chromatography, Liquid; D018920:Coculture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D005057:Eukaryotic Cells / Q000201:enzymology*; D006801:Humans; D053719:Tandem Mass Spectrometry; D041241:Vibrio cholerae O1 / Q000254:growth & development / Q000502:physiology*
chemical_list: D002374:Catalase
keywords: Catalase; FCVC; VBNC; Vibrio cholerae; factor converting VBNC to culturable