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31.
Free-radical polymerization of acrylamide by manganese peroxidase produced by the white-rot basidiomycete Bjerkandera adusta 总被引:1,自引:0,他引:1
Kenji Iwahara Mio Hirata Yoichi Honda Takashi Watanabe Masaaki Kuwahara 《Biotechnology letters》2000,22(17):1355-1361
Acrylamide was polymerized to give polyacrylamide using manganese peroxidase (MnP) produced by the basidiomycete Bjerkandera adusta. The molecular weight of the polymer synthesized by MnP was 155000, higher than those obtained with other reaction systems using horseradish peroxidase and a redox initiator. The 13C-NMR spectrum showed that polyacrylamide was atactic. Electron spin resonance analysis revealed that 2,4-pentanedione added as an initiator was first oxidized to generate a carbon-centered radical, which initiated radical additive polymerization of acrylamide. 相似文献
32.
Biological activities caused by far-infrared radiation 总被引:4,自引:0,他引:4
Contrary to previous presumption, accumulated evidence indicates that far-infrared rays are biologically active. A small ceramic disk that emist far-infrared rays (4–16 m) has commonly been applied to a local spot or a whole part of the body for exposure. Pioneering attempts to experimentally analyze an effect of acute and chronic radiation of far-infrared rays on living organisms have detected a growth-promoting effect in growing rats, a sleep-modulatory effect in freely behaving rats and an insomiac patient, and a blood circulation-enhancing effect in human skin. Question-paires to 542 users of far-infrared radiator disks embedded in bedelothes revealed that the majority of the users subjectively evaluated an improvement of their health. These effects on living organisms appear to be non-specifically triggered by an exposure to far-infrared rays, which eventually induce an increase in temperature of the body tissues or, more basically, an elevated motility of body fluids due to decrease in size of water clusters. 相似文献
33.
Vacuolar protein sorting in fission yeast: cloning, biosynthesis, transport, and processing of carboxypeptidase Y from Schizosaccharomyces pombe. 总被引:2,自引:0,他引:2 下载免费PDF全文
M Tabuchi O Iwaihara Y Ohtani N Ohuchi J Sakurai T Morita S Iwahara K Takegawa 《Journal of bacteriology》1997,179(13):4179-4189
PCR was used to isolate a carboxypeptidase Y (CPY) homolog gene from the fission yeast Schizosaccharomyces pombe. The cloned S. pombe cpy1+ gene has a single open reading frame, which encodes 950 amino acids with one potential N-glycosylation site. It appears to be synthesized as an inactive pre-pro protein that likely undergoes processing following translocation into appropriate intracellular organelles. The C-terminal mature region is highly conserved in other serine carboxypeptidases. In contrast, the N-terminal pro region containing the vacuolar sorting signal in CPY from Saccharomyces cerevisiae shows fewer identical residues. The pro region contains two unusual repeating sequences; repeating sequence I consists of seven contiguous repeating segments of 13 amino acids each, and repeating sequence II consists of seven contiguous repeating segments of 9 amino acids each. Pulse-chase radiolabeling analysis revealed that Cpy1p was initially synthesized in a 110-kDa pro-precursor form and via the 51-kDa single-polypeptide-chain intermediate form which has had its pro segment removed is finally converted to a heterodimer, the mature form, which is detected as a 32-kDa protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Like S. cerevisiae CPY, S. pombe Cpy1p does not require the N-linked oligosaccharide moiety for vacuolar delivery. To investigate the vacuolar sorting signal of S. pombe Cpy1p, we have constructed cpy1+-SUC2 gene fusions that direct the synthesis of hybrid proteins consisting of N-terminal segments of various lengths of S. pombe Cpy1p fused to the secreted enzyme S. cerevisiae invertase. The N-terminal 478 amino acids of Cpy1 are sufficient to direct delivery of a Cpy1-Inv hybrid protein to the vacuole. These results showed that the pro peptide of Cpy1 contains the putative vacuolar sorting signal. 相似文献
34.
Recent studies have shown that lysine side-chain NH3 + groups are excellent probes for NMR investigations of dynamics involving hydrogen bonds and ion pairs relevant to protein function. However, due to rapid hydrogen exchange, observation of 1H-15N NMR cross peaks from lysine NH3 + groups often requires use of a relatively low temperature, which renders difficulty in resonance assignment. Here we present an effective strategy to assign 1H and 15N resonances of NH3 + groups at low temperatures. This strategy involves two new 1H/13C/15N triple-resonance experiments for lysine side chains. Application to a protein-DNA complex is demonstrated. 相似文献
35.
The primary structures of the O-glycosidically linked oligosaccharides isolated from glycoproteins GP I and GP II of Fusarium sp. M7-1 were established. The oligosaccharides released by alkaline borohydride treatment from the glycoproteins were purified by Bio-Gel P-4 and HPLC. This approach resulted in one monosaccharide and seven oligosaccharides. Their primary structures were resolved mainly by NMR spectrometry in combination with methylation mass spectrometry and fast atom bombardment mass spectrometry. The following structures have been determined. [formula: see text]. 相似文献
36.
Two novel decarboxylase genes play a key role in the stereospecific catabolism of dehydrodiconiferyl alcohol in Sphingobium sp. strain SYK‐6 下载免费PDF全文
Sphingobium sp. strain SYK‐6 is able to use a phenylcoumaran‐type biaryl, dehydrodiconiferyl alcohol (DCA), as a sole source of carbon and energy. In SYK‐6 cells, the alcohol group of the B‐ring side chain of DCA was first oxidized to the carboxyl group, and then the alcohol group of the A‐ring side chain was oxidized to generate 5‐(2‐carboxyvinyl)‐2‐(4‐hydroxy‐3‐methoxyphenyl)‐7‐methoxy‐2,3‐dihydrobenzofuran‐3‐carboxylate (DCA‐CC). We identified phcF, phcG and phcH, which conferred the ability to convert DCA‐CC into 3‐(4‐hydroxy‐3‐(4‐hydroxy‐3‐methoxystyryl)‐5‐methoxyphenyl)acrylate (DCA‐S) in a host strain. These genes exhibited no significant sequence similarity with known enzyme genes, whereas phcF and phcG, which contain a DUF3237 domain of unknown function, showed 32% amino acid sequence identity with each other. The DCA‐CC conversion activities were markedly decreased by disruption of phcF and phcG, indicating that phcF and phcG play dominant roles in the conversion of DCA‐CC. Purified PhcF and PhcG catalysed the decarboxylation of the A‐ring side chain of DCA‐CC, producing DCA‐S, and showed enantiospecificity towards (+)‐ and (–)‐DCA‐CC respectively. PhcF and PhcG formed homotrimers, and their Km for DCA‐CC were determined to be 84 μM and 103 μM, and Vmax were 307 μmol?min?1?mg?1 and 137 μmol?min?1?mg?1 respectively. In conclusion, PhcF and PhcG are enantiospecific decarboxylases involved in phenylcoumaran catabolism. 相似文献
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39.
K Takegawa A Kondo H Iwamoto K Fujiwara Y Hosokawa I Kato K Hiromi S Iwahara 《Biochemistry international》1991,25(1):181-190
The primary structure of the N-linked sugar chains of glucose oxidase from Aspergillus niger was investigated. These sugar chains were released from the polypeptide backbone by hydrazinolysis, and the reducing ends of the sugar chains were pyridylaminated. HPLC of the pyridylamino sugar chains with an amide-silica column showed at least seven sugar chain peaks. Chemical and exoglycosidase digestion and 400 lMHz H-NMR studies of the sugar chains of lower molecular weight showed that these were novel oligomannose-type sugar chains, (Man)5-7 (GlcNAc)2, with the structure: +/- Man alpha 1----3Man alpha 1----3(Man alpha 1----6)Man alpha 1----6(+/- Man alpha 1----3Man alpha 1---3)Man )Man beta 1----4GlcNAc beta 1----4GlcNAc. 相似文献
40.
Endo-beta-N-acetylglucosaminidase from Arthrobacter protophormiae was tested for its capacity to release N-linked sugar chains from native yeast invertase. The enzyme liberated about 80% of the sugar chains from the native invertase. Deglycosylated invertase was digested by chymotrypsin or pepsin, and twelve N-acetylglucosamine-containing glycopeptides were isolated. The amino acid sequences of these glycopeptides were analyzed by a protein sequencer, and the elution position of 4-L-aspartylglycosylamine was directly identified by conventional sequencing. The endo-beta-N-acetylglucosaminidase was found to remove mainly nine sugar chains from native invertase. 相似文献