PMID- 16950902
OWN - NLM
STAT- MEDLINE
DCOM- 20070123
LR  - 20191210
IS  - 0099-2240 (Print)
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Linking)
VI  - 72
IP  - 11
DP  - 2006 Nov
TI  - Application of pseudomurein endoisopeptidase to fluorescence in situ 
      hybridization of methanogens within the family Methanobacteriaceae.
PG  - 6907-13
AB  - In situ detection of methanogens within the family Methanobacteriaceae is 
      sometimes known to be unsuccessful due to the difficulty in permeability of 
      oligonucleotide probes. Pseudomurein endoisopeptidase (Pei), a lytic enzyme that 
      specifically acts on their cell walls, was applied prior to 16S rRNA-targeting 
      fluorescence in situ hybridization (FISH). For this purpose, pure cultured 
      methanogens within this family, Methanobacterium bryantii, Methanobrevibacter 
      ruminantium, Methanosphaera stadtmanae, and Methanothermobacter 
      thermautotrophicus together with a Methanothermobacter 
      thermautotrophicus-containing syntrophic acetate-oxidizing coculture, 
      endosymbiotic Methanobrevibacter methanogens within an anaerobic ciliate, and an 
      upflow anaerobic sludge blanket (UASB) granule were examined. Even without the 
      Pei treatment, Methanobacterium bryantii and Methanothermobacter 
      thermautotrophicus cells are relatively well hybridized with oligonucleotide 
      probes. However, almost none of the cells of Methanobrevibacter ruminantium, 
      Methanosphaera stadtmanae, cocultured Methanothermobacter thermautotrophicus, and 
      the endosymbiotic methanogens and the cells within UASB granule were hybridized. 
      Pei treatment was able to increase the probe hybridization ratio in every 
      specimen, particularly in the specimen that had shown little hybridization. 
      Interestingly, the hybridizing signal intensity of Methanothermobacter 
      thermautotrophicus cells in coculture with an acetate-oxidizing H(2)-producing 
      syntroph was significantly improved by Pei pretreatment, whereas the probe was 
      well hybridized with the cells of pure culture of the same strain. We found that 
      the difference is attributed to the differences in cell wall thicknesses between 
      the two culture conditions. These results indicate that Pei treatment is 
      effective for FISH analysis of methanogens that show impermeability to the probe.
FAU - Nakamura, Kohei
AU  - Nakamura K
AD  - Institute for Biological Resources and Functions, National Institute of Advanced 
      Industrial Science and Technology, 1-1-1 Higashi, Tsukuba 305-8566, Japan.
FAU - Terada, Takeshi
AU  - Terada T
FAU - Sekiguchi, Yuji
AU  - Sekiguchi Y
FAU - Shinzato, Naoya
AU  - Shinzato N
FAU - Meng, Xian-Ying
AU  - Meng XY
FAU - Enoki, Miho
AU  - Enoki M
FAU - Kamagata, Yoichi
AU  - Kamagata Y
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060901
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Culture Media)
RN  - 0 (Sewage)
RN  - 0 (peptidoglycan endopeptidase)
RN  - EC 3.4.- (Endopeptidases)
SB  - IM
MH  - Animals
MH  - Ciliophora/growth & development/microbiology
MH  - Culture Media
MH  - Endopeptidases/genetics/*metabolism
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Methanobacteriaceae/*classification/genetics/growth & development/*metabolism
MH  - Sewage/microbiology
PMC - PMC1636154
EDAT- 2006/09/05 09:00
MHDA- 2007/01/24 09:00
PMCR- 2007/03/01
CRDT- 2006/09/05 09:00
PHST- 2006/09/05 09:00 [pubmed]
PHST- 2007/01/24 09:00 [medline]
PHST- 2006/09/05 09:00 [entrez]
PHST- 2007/03/01 00:00 [pmc-release]
AID - AEM.01499-06 [pii]
AID - 1499-06 [pii]
AID - 10.1128/AEM.01499-06 [doi]
PST - ppublish
SO  - Appl Environ Microbiol. 2006 Nov;72(11):6907-13. doi: 10.1128/AEM.01499-06. Epub 
      2006 Sep 1.