Pyrococcus woesei

DIP is a complex solute that was first noted as a major solute in Pyrococcus woesei (Scholz et al., 1992) and Methanococcus igneus (Ciulla et al., 1994). In P. woesei, DIP accumulates in quantities comparable to intracellular... 

Scholz, S., Sonnenbichler, J., Schafer, W., and Hensel, R., 1992, Di-myo-inositol-1,1 -phosphate A new inositol phosphate isolated from Pyrococcus woesei. FEBS Lett. 306 239-242. 

Further studies indicate that an ADP-forming acetyl-CoA synthetase is also operative in other extremely thermophilic archaea Pyrococcus woesei, Thermococcus celer, Hyperthermus butylicus, Desulfurococcus amylolyticus), which form acetate as end product of their fermentation [305]. In contrast, in acetate forming (eu)bacteria, acetate formation from acetyl-CoA and the synthesis of ATP from ADP and Pj are catalyzed by two enzymes phosphate acetyltransferase and acetate kinase. This holds true for the extremely thermophilic (eu)bacterium. Thermotoga maritima[3Q5], which ferments... 

Cell extracts of Pyrococcus contain low activities of fructose-1,6-bisphosphate aldolase [295] and NAD(P)-dependent glyceraldehyde-3-phosphate dehydrogenase [301, 308a], which probably have anabolic functions (see ref [308b]). Glyceraldehyde-3-phosphate dehydrogenase has been purified from Pyrococcus woesei [308a] (see Chapter 7 by Hensel in this volume). 

Data for Pyrococcus woesei are from Cammarano et al. (unpublished) ) data for Desuljurococcus mobilis are from Ceccarelli, E., et al. (unpublished) data for T. celer are from Auer, J. (unpublished). 

Abbreviations Asa, Artemia salina Dme, Drosophila melanogaster Hsa, Homo sapiens Egr, Euglena gracilis, Ddi, Dictyostelium discoideum Sso, S. solfataricus Tee, Thermococcus celer Pwo, Pyrococcus woesei Mva, M. vannielii Hma, Halobacterium marismortui Tac, Thermoplasma acidophilum Tma, Thermotoga marilima Tth, Thermus thermophilus Eco, E. coli Mlu, Micrococcus luteus, Spl, Spirulina platensis Ech, E. gracilis chloroplast Smt, Saccharomyces cerevisiae mitochondria. 

Pyrococcus woesei[10°1 100 6.0 90 Purified/cloned/cell associated... 

Pullulanase Type 11 pullulan, branched polysaccharides [endoacting a-1,6 in pullulan a-1,6 + a-1,4 in branched poly- and oligosaccharide] maltotriose, linear oligosaccharides B. subtilis C. thermohydrosuljuricum Pyrococcus woesei Desuljurococcus mucosus... 

Pwo polymerase DNA polymerase isolated from pyrococcus woesei, an enzyme with 5 -3 polymerase and 3 -5 exonuclease activity. This DNA polymerase generates blunt-ended DNA fragments. It amplifies DNA targets up to 3 kb. Optimal concentration is 0.5-1.5 units/50 pi reaction volume with 2 mM MgS04. 

DIP was initially identified in Pyrococcus woesei and Methanococcus igneusP Later, this solute was detected in other hyperthermophilic archaea, namely in Pyrodictium occultum, and in Pyrococcus and Thermococcus spp. In several species of the Thermococcales large increases in the levels of DIP are observed at growth temperatures above the optimum, leading to the view that this solute has a thermoprotective role in these oiganisms. ... 

Two different pathways for the synthesis of DIP have been investigated in Pyrococcus woesei and Methanococcus igneus. In P. woesei the synthesis was reported to proceed via two steps glucose 6-phosphate is converted into... 

Inositol Phosphates and Derivatives.-The synthesis of Dwi v-inoatol 3,4,S,6-tetrakisphosphate as previously reported (Vol. 26, p. 206, ref. 154) was in fact of the enantiomer. A novel inositol phosphate, L,L-di-myo-inositoI-l,r-phosphate has been isolated from Pyrococcus woesei and its structure confirmed by synthesis. ... 

Figure 1 Purification of Pyrococcus woesei NADP-dependent GluDH by two sequential Red Sepharose CL-4B column chromatographies. (A) Elution profile of the first chromatography. Crude enzyme (235 mg, 63.5 U, specific activity 0.27, in the 10 mM phosphate buffer, pH 7.2) was applied on the first column and the enzyme (4.41 mg, 44.7 U, specific activity 10.1) was eluted with 20 mM phosphate buffer (pH 8.0) containing 0.5 M NaGl. (B) Elution profile of the second chromatography. Enzyme solution (pH 7.2) dialyzed was applied on the affinity column and GluDH (0.88 mg, 21.4 U, specific activity 24.3, yield 33.7%) was eluted with a linear gradient of NADP concentration (0-2 mM) in the presence of 10 mM L-glutamate [36].

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