Production of Pyrroloquinoline Quinone by Using Methanol-Utilizing Bacteria

A large number of methanol-utilizing bacteria were screened for extracellular production of pyrroloquinoline quinone (PQQ) by using methanol as the carbon and energy sources. Of the bacteria selected, Hyphomicrobium sp. strain TK 0441 was examined for PQQ production by using a jar fermentor. The amount of PQQ in the broth and the level of methanol dehydrogenase activity in the cells were increased by simply decreasing the amount of Fe added to the medium. On the other hand, extracellularly produced protein which interfered with the purification of PQQ was decreased by simply increasing the amount of Mg added to the medium. A suitable medium that contained 1 μg of Fe per ml, 150 μg of Mg per ml, and trace elements was developed. In this medium, the production of PQQ reached approximately 1 mg/ml and protein formation was low.     

Gene cloning of Porphyromonas gingivalis specific antigens recognized by serum of adult periodontitis patient.

1. Porphyromonas gingivalis is believed an important pathogen of adult periodontitis. A gene library of P. gingivalis 381 was constructed in lambda phage vector L47.1. The library was probed with serum obtained from patients of severe adult periodontitis. Two clones, lambda MDBG101 and lambda MDBG103 which were expressed, 200 and 160 kDa respectively, were selected and further studied. 2. The expressed antigens in these two clones were also reacted with rabbit antiserum against whole cells, capsular fraction and cell surface fraction of P. gingivalis. 3. Genes coding protein antigens in lambda MDBG101 and lambda MDBG103 were subcloned into high-copy-number plasmid vector pACYC184 and subclones obtained were designated as MD101 and MD103. Recombinant plasmids, pMD101 and pMD103, differed in their restriction endonuclease digestion. 4. Immunodiffusion analysis showed that cloned proteins from MD101 and MD103 reacted with antiserum against P. gingivalis but did not react with antiserum against Prevotella intermedia, Prevotella loescheii and Prevotella asaccharolyticus. 5. These data suggest that P. gingivalis species-specific antigens has been successfully cloned and expressed in Escherichia coli. Since these cloned specific antigens were recognized by adult periodontitis patient sera, the recombinant antigen will be useful material for the development of serodiagnosis of P. gingivalis infection in adults periodontitis.     

Effects of iloprost,a PGI2 derivative,on ischemic myocardial energy and carbohydrate metabolism in dogs

Effects of iloprost, which is a stable prostacyclin analogue, on the ischemic myocardium were examined in the open-chest dog heart, in terms of biochemical parameters. Ischemia was initiated by ligating the left anterior descending coronary artery. When the coronary artery was ligated for 3 min, the levels or glycogen, fructose-1,6-diphosphate (FDP), adenosine triphosphate and creatinephosphate decreased, and the levels of glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), lactate, adenosine diphosphate and adenosine monophosphate increased. During ischemia, therefore, energy charge potential was significantly decreased from 0.89±0.01 to 0.82±0.01, and ([G6P]+[F6P])/[FDP] and [lactate]/[pyruvate] ratios were significantly increased from 1.75±0.30 to 29.05±5.70 and 13±3 to 393±112, respectively. Iloprost (0.1, 0.3, or 1 g·kg^–1) was injected intravenously 5 min before the onset of ischemia. Iloprost (0.1, 0.3, and 1 g·kg^–1) reduced the ischemia-induced decrease in energy charge potential to 94, 74, and 86%, respectively, the increase in ([G6P]+[F6P]/[FDP] to 38, 29, 32%, respectively, and the increase in [lactate]/[pyruvate] to 67, 45, 65%, respectively. These results suggest that iloprost lessens the myocardial metabolic derangements produced by ischemia, and the most potent effect was obtained at the dose of 0.3 g·kg^–1.     

A 718-kb DNA Sequence of the Escherichia coli K-12 Genome Corresponding to the 12.7-28.0 min Region on the Linkage Map

The 718,122 base pair (bp) sequence of the Escherichia coliK-12 genome corresponding to the region from 12.7 to 28.0 minuteson the genetic map is described. This region contains at least682 potential open reading frames, of which 278 (41%) have beenpreviously identified, 147 (22%) were homologous to other knowngenes, 138 (20%) are identical or similar to the hypotheticalgenes registered in databases, and the remaining 119 (17%) didnot show a significant similarity to any other gene. In thisregion, we assigned a cluster of cit genes encoding multienzymecitrate lyase, two clusters of fimbrial genes and a set of lysogenicphage genes encoding integrase, excisionase and repressor inthe e14 genetic element. In addition, a new valine tRNA gene,designated valZ, and a family of long directly repeated sequences,LDR-A, -B and -C, were found.     

Clustered Genes for Galactose Metabolism from Streptococcus mutans Cloned in Escherichia coli

DNA cloned into Escherichia coli from a serotype c strain of Streptococcus mutans allowed a galKTE mutant to utilize galactose for growth. However, the DNA does not appear to encode enzymes of the Leloir pathway used by E. coli, but rather appears to encode enzymes of the tagatose phosphate pathway.     

H2O2 activates ryanodine receptor but has little effect on recovery of releasable Ca2+ content after fatigue.

We studied whether hydrogen peroxide (H(2)O(2)) at 相似文献   

Steroidogenesis in the zona glomerulosa of the adrenal cortex II. Distribution of cytochrome P-450 in the zona glomerulosa of the bovine adrenal cortex

Microsomes were obtained from the zona glomerulosa of the bovine adrenal cortex. Contamination of microsomes with other cellular organelles was examined using various marker enzymes and the electron microscope. Distribution of cytochrome P-450 in the zona glomerulosa was studied using various fractions including microsomes, described above, and mitochondria. The amount of cytochrome P-450 in mitochondria and in microsomes was determined to be 0.73 and 0.32 nmol/mg protein, respectively. The CO difference spectrum was affected not only by the concentration of added deoxycholate but also by the incubation time after addition. Approximately 40–50% of cytochrome P-450 in the samples was converted to cytochrome P-420 within 20–30 sec of incubation with deoxycholate.The content of RNA, phospholipids, and cytochromeb ₅ in microsomes obtained from the zona glomerulosa is also evaluated in comparison to that in microsomes obtained from the zona fasciculoreticularis.     

Purification and mode of action of a microsomal endoribonuclease from rat liver

An endoribonuclease has been purified nearly to homogeneity from rat liver microsomes, and its mode of action and general properties were studied. The enzyme had an apparent molecular weight of 58 000, as estimated by both gel filtration and SDS-polyacrylamide gel electrophoresis and produced oligonucleotides from poly(A), poly(U) and poly(C). No mononucleotide was obtained by the enzymatic hydrolysis of the above substrates. The enzyme made endonucleolytic cleavages which generated 5'-phosphate-terminated oligonucleotides. It was suggested that the existence of at least (Ado5'P)2 residues at both sides of the cleavage bond was necessary for the action of the endoribonuclease. Divalent cations (Mg2+ or Mn2+) were required for the enzymatic activity, while K+ inhibited the enzyme. Spermine stimulated the enzymatic activity in the presence of 1 mM Mg2+.     

Identification and synthesis of a heptapeptide in uremic fluid

A heptapeptide isolated from uremic fluid was synthesized by conventional method. The total amino acid sequence of this peptide was deduced as follows: H-His-Pro-Ala-Glu-Asn-Gly-Lys-OH. Structural similarity was soon realized between this peptide and heptapeptde moiety corresponding to position 13 through 19 of β₂-microglobulin.