Senoh Mitsutoshi, Hamabata Takashi, Takeda Yoshifumi
Collaborative Research Center of Okayama University for Infectious Diseases in India, Okayama University, Kolkata, India.
Research Institute, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan.
Microbiologyopen. 2015 Aug;4(4):589-96. doi: 10.1002/mbo3.264. Epub 2015 May 13.
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.
在我们之前的工作中,我们证明了活的但不可培养的(VBNC)霍乱弧菌O1和O139通过与真核细胞共培养可转化为可培养状态。此外,我们从真核细胞系HT-29中分离出一种将VBNC霍乱弧菌转化为可培养状态的因子(FCVC)。在本研究中,我们通过包括UNO Q-6阴离子交换、Bio-Scale CHT2-1羟基磷灰石和Superdex 200 10/300 GL的连续柱色谱法纯化了FCVC。通过SDS-PAGE证明了纯化后的FCVC的均一性。纳升液相色谱串联质谱(Nano-LC MS/MS)分析表明,纯化后的FCVC是一种人过氧化氢酶。来自HT-29细胞的过氧化氢酶mRNA的RNA干扰敲低实验以及用过氧化氢酶抑制剂3-氨基-1,2,4-三唑处理纯化后的FCVC证实了FCVC是一种过氧化氢酶。本文讨论了过氧化氢酶在将人肠道中的VBNC霍乱弧菌转化为可培养状态中可能发挥的作用。