Study on the conversion from VBNC to culturable state in Vibrio cholerae

2016 Fiscal Year Final Research Report

• Project/Area Number: 26460543
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bacteriology (including mycology)
• Research Institution: National Center for Global Health and Medicine
• Principal Investigator: Hamabata Takashi 国立研究開発法人国立国際医療研究センター, その他部局等, 細菌感染研究室長 (40311427)
• Co-Investigator(Renkei-kenkyūsha): Senoh Mitsutoshi 国立感染症研究所, 細菌第二部, 主任研究官 (20646624)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: コレラ菌 / カタラーゼ / VBNC / RNAマイクロアレイ

Outline of Final Research Achievements

The viable but nonculturable (VBNC) Vibrio cholerae converts to culturable state by co-culture of with cultured cells. The converting factor (FCVC) was purified from HT-29 cells through several kinds of column chromatography. The final purified single band on SDS-PAGE was analyzed by nano-LC MS/MS and turned out to be human catalase. Both catalase RNAi analysis of HT-29 cells and catalase inhibitor 3-amino-1,2,4-triazole greatly reduced the converting activity of crude cell extract, confirming that FCVC is catalase.
To identify the genes that involve in the conversion, VBNC V. cholerae was collected by centrifugation, treated with catalase and bacterial RNA was purified on several different time points. Since RNA microarray analysis revealed too many genes responded to catalase, negative control RNA should be compared. However, low conversion was always seen even without catalase, which reason turned out to be the condensation of bacteria by centrifugation.

Free Research Field

細菌学