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81.
Phylogenetic Analysis of Bacteria Preserved in a Permafrost Ice Wedge for 25,000 Years 总被引:1,自引:0,他引:1
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Taiki Katayama Michiko Tanaka Jun Moriizumi Toshio Nakamura Anatoli Brouchkov Thomas A. Douglas Masami Fukuda Fusao Tomita Kozo Asano 《Applied microbiology》2007,73(7):2360-2363
Phylogenetic analysis of bacteria preserved within an ice wedge from the Fox permafrost tunnel was undertaken by cultivation and molecular techniques. The radiocarbon age of the ice wedge was determined. Our results suggest that the bacteria in the ice wedge adapted to the frozen conditions have survived for 25,000 years. 相似文献
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Prostaglandin E2 synthase (PGES) catalyzes the isomerization of PGH2 to PGE2. PGES type 2 (mPGES-2) is a membrane-associated enzyme, whose N-terminal section is apparently inserted into the lipid bilayer. Both intact and N-terminal truncated enzymes have been isolated and have similar catalytic activity. The recombinant N-terminal truncated enzyme purified from Escherichia coli HB101 grown in LB medium containing delta-aminolevulinate and Fe(NO3)3 has a red color, while the same enzyme purified from the same E. coli grown in minimal medium has no color. The red-colored enzyme has been characterized by mass, fluorescence, and EPR spectroscopies and X-ray crystallography. The enzyme is found to contain bound glutathione (GSH) and heme. GSH binds to the active site with six H-bonds, while a heme is complexed with bound GSH forming a S-Fe coordination bond with no polar interaction with mPGES-2. There is a large open space between the heme and the protein, where a PGH2 might be able to bind. The heme dissociation constant is 0.53 microM, indicating that mPGES-2 has relatively strong heme affinity. Indeed, expression of mPGES-2 in E. coli stimulates heme biosynthesis. Although mPGES-2 has been reported to be a GSH-independent PGES, the crystal structure and sequence analysis indicate that mPGES-2 is a GSH-binding protein. The GSH-heme complex-bound enzyme (mPGES-2h) catalyzes formation of 12(S)-hydroxy-5(Z),8(E),10(E)-heptadecatrienoic acid and malondialdehyde from PGH2, but not formation of PGE2. The following kinetic parameters at 37 degrees C were determined: KM = 56 microM, kcat = 63 s-1, and kcat/KM = 1.1 x 10(6) M-1 s-1. They suggest that mPGES-2h has significant catalytic activity for PGH2 degradation. It is possible that both GSH-heme complex-free and -bound enzymes are present in the same tissues. mPGES-2 in heme-rich liver is most likely to become the form of mPGES-2h and might be involved in degradation reactions similar to that of cytochrome P450. Since mPGES-2 is an isomerase and mPGES-2h is a lyase, mPGES-2 cannot simply be classified into one of six classes set by the International Union of Biochemistry and Molecular Biology. 相似文献
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Proton (H+) conductive pathways are suggested to play roles in the regulation of intracellular pH. We characterized temperature-sensitive whole cell currents in mouse bone marrow–derived mast cells (BMMC), immature proliferating mast cells generated by in vitro culture. Heating from 24 to 36°C reversibly and repeatedly activated a voltage-dependent outward conductance with Q10 of 9.9 ± 3.1 (mean ± SD) (n = 6). Either a decrease in intracellular pH or an increase in extracellular pH enhanced the amplitude and shifted the activation voltage to more negative potentials. With acidic intracellular solutions (pH 5.5), the outward current was detected in some cells at 24°C and Q10 was 6.0 ± 2.6 (n = 9). The reversal potential was unaffected by changes in concentrations of major ionic constituents (K+, Cl−, and Na+), but depended on the pH gradient, suggesting that H+ (equivalents) is a major ion species carrying the current. The H+ current was featured by slow activation kinetics upon membrane depolarization, and the activation time course was accelerated by increases in depolarization, elevating temperature and extracellular alkalization. The current was recorded even when ATP was removed from the intracellular solution, but the mean amplitude was smaller than that in the presence of ATP. The H+ current was reversibly inhibited by Zn2+ but not by bafilomycin A1, an inhibitor for a vacuolar type H+-ATPase. Macroscopic measurements of pH using a fluorescent dye (BCECF) revealed that a rapid recovery of intracellular pH from acid-load was attenuated by lowering temperature, addition of Zn2+, and depletion of extracellular K+, but not by bafilomycin A1. These results suggest that the H+ conductive pathway contributes to intracellular pH homeostasis of BMMC and that the high activation energy may be involved in enhancement of the H+ conductance. 相似文献
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Hirofumi Nakano Fusao Tomita Takeo Suzuki 《Bioscience, biotechnology, and biochemistry》2013,77(12):2471-2475
For the purpose of improving the productivity of corynecins (chloramphenicol analogs), the mutation study of the producer, Corynebacterium hydrocarboclastus, was concerned in this investigation with the isolation of glycolipids defective strains. The mutant KY 8834 possessed glycolipids less than one-third to one-tenth of the parent. Productivity of corynecins of the mutant was better than that of the parent, In addition, this character of the mutant was advantageous for the stirring fermentation, because aggregation of cells was remarkably diminished unlike the case of the parental strain.Another mutant was isolated by mutagenesis of glycolipids defective mutant, KY 8834. The mutant, KY 8835, produced dominantly corynecin I (less toxic homolog to the producer strain). Thus, the productivity of corynecins was elevated to about two-fold of that of strain KY8834. 相似文献
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Fusao Tomita Toshihide Nakanishi Takeo Suzuki 《Bioscience, biotechnology, and biochemistry》2013,77(2):293-299
Extracellular accumulation of high molecular weight DNA was further studied using Pseudomonas species. More efficient production was obtained by the use of glucose-grown seed culture and by controlling the broth-pH at around 6.0 for first 24 hr and then around 8.0 during the fermentation. The maximum yield was 5 to 6 g per liter of the broth culture, which corresponded to 10-fold of that reported in the previous work.Purified DNA (4 × 106 daltons) was obtained successfully by applying an aqueous biphase system of dextran-polyethyleneglycol and dextranase.Significant release of DNA occurred only with cell lysis of H-paraffin-grown bacteria. The primary cause of rapid lysis was explained by the exhaustion of cellular glucose pool. Relation of DNA accumulation to the effect of rhamnolipids on cell membrane was also investigated. 相似文献
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Tadamichi Morisaka Mai Sakai Kazunobu Kogi Akane Nakasuji Kasumi Sakakibara Yuria Kasanuki Motoi Yoshioka 《PloS one》2013,8(8)
Spontaneous ejaculation, which is defined as the release of seminal fluids without apparent sexual stimulation, has been documented in boreoeutherian mammals. Here we report spontaneous ejaculation in a wild Indo-Pacific bottlenose dolphin (Tursiops aduncus), and present a video of this rare behavior. This is the first report of spontaneous ejaculation by an aquatic mammal, and the first video of this behavior in animals to be published in a scientific journal. 相似文献
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Hirofumi Nakano Fusao Tomita Takeo Suzuki 《Bioscience, biotechnology, and biochemistry》2013,77(12):2505-2509
In the biosynthesis of corynecins by Corynebacterium hydrocarboclastus, it appeared that shikimic acid was one of the efficient precursors, where shikimic acid-U-14C was incorporated into corynecins in the yield of approximately 15%. Analyses of degradation products of labeled corynecins demonstrated that shikimic acid was incorporated specifically into aromatic ring of corynecins.The incorporation of shikimic acid was inhibited by several aromatic amines such as p-aminophenylserinol-N-propionamide, although the uptake of shikimic acid was not affected, suggesting that biosynthesis of corynecins might be regulated by p-aminophenyl intermediates. Furthermore, p-ammophenylethylalcohol was found to be a potent inhibitor of biosynthesis of corynecins. In contrast, corynecins and other p-nitro-phenyl derivatives, aromatic amino acids and vitamins related to shikimic acid pathway did not inhibit the biosynthesis of corynecins from shikimic acid. 相似文献