Introduction Thermotogales 1987 Thermotoga Aquificales Thermotogales 2005 Thermotogales Thermotoga Thermosipho Fervidobacterium Geotoga Petrotoga Marinitoga 2005 T. maritima Thermotoga Thermotogales 1986 T. neapolitana T. petrophila T. naphtophila T. thermarum T. elfii, T. subterranea T. hypogea T. lettingae) Thermotoga 2005 Fervidobacterium Fervidobacterium pennivorans 1996 Thermosipho T. africanus T. melanesiensis 1997 1989 Thermotogales Thermotogales Thermotoga 1995 1992 1989 1997 1988 1986 T. maritima 1992 Thermotogales Materials and methods Cultures 1 2 2 4 2 4 2 4 3 2 2 http://www.dsmz.de 2 2 2 2 3 2 2 2 2 4 2 2 2 4 2 4 3 2 2 2 2 1 T. lettingae Table 1 Thermotogales Bacterial species Source for the isolate Optimal growth temperature (°C) Reference DSM strain number Culture conditions a T (°C)  Thermotoga maritima Anaerobic marine sediment, Italy  80 1986 3109 A 80, 55 T. neapolitana Submarine hot spring, Italy  85 1988 4359 A 80 T. elfii Oil-producing well, Africa  65 1995 9442 A 65 T. lettingae Sulfate-reducing bioreactor, The Netherlands  65 2002 14385 A 65 T. hypogea Oil-producing well, Africa  70 1997 11164 B 70 Thermosipho sp. Spring, New Zealand  65  6568 B 65 T. africanus Marine hydrothermal spring, Obock  75 1989 5309 B 75 T. melanesiensis Deep sea hydrothermal vent, Pacific Ocean  70 1997 12029 B 70 Fervidobacterium pennivorans Hot mud of spa, Italy  65 1997 9078 B 65 a Materials and method Lipid analysis 3 1972 The cell residue after solvent extraction was, after addition of a known amount of the internal standard, 2-methylheneicosane, hydrolysed with 1 N KOH/MeOH (96%) by refluxing for 1 h. The hydrolysate was neutralized with 2 N HCl/MeOH and after addition of water extracted with DCM. This fraction was methylated, chromatographed over silica and silylated as described above and the resulting TLF-2 fraction was analyzed by GC and GC–MS. In specific cases alkaline hydrolysis was also performed with an aliquot of the solvent extract following the same procedure as described above. This resulted in a fraction named TLF-1-OH which was also analyzed by GC and GC–MS. 2 3  GC and GC–MS d f HPLC/APCI–MS 2000 −1 Results Thermotogales Thermotoga T. maritima T. lettingae T. elfii T. neapolitana T. hypogea Thermosipho T. africanus T. melanesiensis T. sp. Fervidobacterium F. pennivorans 1 Composition of total lipid fractions 1 T. maritima n 16 n 14 1 2 2 1997 1979 1 2 3 30 32 2 F. pennivorans 34 O 3 O 4 5 6 5 5 1988 5 1988 5 3 6 5  3 5 6 Thermotogales 2 1 2 T. lettingae 4 7  8 9 10 1980 Thermotoga O Fig. 1 a b c d Thermotoga maritima F. pennivorans pennivorans 2 FA std  Fig. 2 Thermotogales  Fig. 3 Mass spectra (subtracted for background) of derivatised diabolic acids and diabolic acid glycerol ethers  Table 2 Thermotogales Bacterial species a b 14:0 16:0 18:0 1 2 3 4 5 6 Thermotoga maritima  TLF-1 4.2 44.2 1.1 21.3 1.7 9.1 5.1 10.6 2.7 TLF-2 1.7 25.9 0.9 38.7 4.2 2.2 1.0 17.6 7.8 T. maritima  TLF-1 7.5 54.7 0.5 14.3 2.3 11.1 2.5 6.6 1.2 TLF-2 0.7 15.0 0.4 48.4 4.6 0.7 0.2 23.8 6.2 T. neapolitana TLF-1 4.6 61.3 2.8 17.3 1.3 3.7 0.7 7.1 1.2 TLF-2 1.9 46.5 2.8 26.4 2.1 0.6  15.8 3.9 T. elfii TLF-1 3.5 61.0 4.0 28.2 2.6 0.5 0.1 0.2  TLF-2 1.2 20.0 1.2 70.7 6.4 0.2  0.3  T. lettingae  TLF-1 3.0 37.2 5.9 24.6 5.2 6.9 3.9 8.3 5.0 TLF-2 0.8 11.6 1.0 48.3 7.8 2.2 0.9 18.7 8.7 T. lettingae  TLF-1-OH 6.3 26.4 1.1 32.6 8.8 4.8 5.8 8.5 5.6 TLF-2 0.5 7.4 0.3 57.6 14.1 0.4 0.2 11.8 7.7 T. hypogea TLF-1 4.8 54.5 2.3 14.1 2.3 7.4 3.5 8.1 3.0 TLF-2 0.5 4.5 1.0 47.4 1.5 0.6 0.1 33.3 11.1 Thermosipho sp. TLF-1 7.9 51.5 28.0 12.6      TLF-2 2.6 16.0 5.0 66.8 9.6     T. africanus TLF-1 4.4 57.6 32.5 2.7      TLF-2 2.5 61.8 8.1 25.4 2.2     T. melanesiensis TLF-1 5.5 62.1 30.8 1.6      TLF-2 5.6 84.7 7.1 2.6      Fervidobacterium. pennivorans TLF-1 1.0 54.1 9.5 35.1 0.3     TLF-2 0.2 13.5 3.0 82.3 1.0     a TLF-1 TLF-2 TLF-OH b 2 Fig. 4 a b c Thermotoga lettingae 2 FA std  2 16:1 18:1 Thermosipho 1 2  5 6 1 4 2 T. lettingae 1 2  5 6 4 4 −1 3 Thermotoga F. pennivorans Thermosipho T. lettingae Table 3 − 1 Thermotogales Bacterial species a b 1 2 5 6 Thermotoga maritima  TLF-1 5.6 0.51 2.7 0.50 TLF-2 4.1 0.40 2.0 0.52 T. lettingae  TLF-1     TLF-1-OH 7.1 1.9 1.8 1.2 TLF-2 1.9 0.46 0.38 0.25 T. hypogea TLF-1 1.0 0.17 0.60 0.22 TLF-2 0.70 0.02 0.49 0.16 Thermosipho sp. TLF-1 0.05    TLF-2 0.26 0.04   T. africanus TLF-1 0.01    TLF-2 0.18 0.02   T. melanesiensis TLF-1     TLF-2 0.01    Fervidobacterium pennivorans TLF-1 7.9 0.07   TLF-2 3.5 0.04   a TLF-1 TLF-2  TLF-1-OH b 2 T. maritima Thermotogales 2000 Materials and methods Thermotoga maritima 5 5 6 + + + 2000 2000 32 70 140 6 1 5 T. maritima 11a 11a 32 30 12a 2 6 2 Fig. 5 a b i Thermotoga maritima b + 70 c + 70 d + 68 e + 70 f + 68 g + 70 h + 68 i + 70 j + 68 Filled blocked 32 32 32 30  Fig. 6 a 70 11a b 70 11b  11c c 70 11d d 70 11e e 70 11f f 70 13d  11b 11c 5 6 11a  6 + + + 11a 32 11b 70 138 7 + 5 + 11 11b 11c 11b 32 30 12b 5 + 5 11b 6 32 11d 70 136 8 + 5 5 32 30  12d 5 5 32 11e 5 5 32 11e 32 30 12e)  6 32 11f 32 30 5 5 6 T. maritima T. lettingae T. lettingae 7 T. maritima 32 11d 7 32 30  12d 7 30 7 30 2 32 1 33 6  35 5  T. lettingae T. maritima 2 32 30  30  7 7 11a T. maritima T. lettingae Fig. 7 a b j Thermotoga lettingae b + 70 c + 68 d + 66 e + 70 f + 68 g + 66 h + 70 i + 68 j + 66 Filled blocked grey 32 32 32 30 30 30  T. maritima T. lettingae 8 + 6 8 7 13d 32 30  16 + 2000 T. lettingae 16  8 8 Fig. 8 a b j Thermotoga lettingae b + 70 c + 68 d + 66 e + 70 f + 68 g + 66 h + 70 i + 68 j + 66 Thermotoga lettingae Black blocked grey 32 32 32 30 30 30  F. pennivorans F. pennivorans 32 11f T. maritima 5 T. lettingae 7 32 34 Discussion Thermotogales 1988 1989 1995 1986 2 Thermotogales 1980 Butyrivibrio Butyrivibrio Thermotogales 2 T. maritima Thermotoga 3 6, 9 11e, 12a 12e, 13a 13e Thermosipho Fervidobacterium 2 F. pennivorans Thermotoga 5 7 8 Thermotoga 9 Thermosipho Fervidobacterium Thermotoga Thermotoga Fig. 9 Thermotogales Escherichia col F. penniforans Fervidobacterium  3 4 1 2 5 6 1 6 1992 2 3 2 2 16:0 1 16:0 2 34 30 2 F. pennivorans T. lettingae Thermosipho sp 18:0 14:0 2 T. lettingae 30 2 F. pennivorans 34 T. maritima 16:0 2002 16:0 Thermotoga Thermotoga 2 T. lettingae 4 1989 2002 1993 Thermotogales T. lettingae 4 5 6 8 Thermotogales Thermotoga Planctomycetes 2002 2005 Aquifex 1992 2001 11a  12a T. maritima 2000 1993 Butyrivibrio 1980 Thermoanaerobacter 1994 Thermotogales 1990 Thermotoga maritima 1999 Thermotogales Thermotogales