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Shuwsei Kamimiya Yoshifumi Itoh Kazuo Izaki Hajime Takahashi 《Bioscience, biotechnology, and biochemistry》2013,77(6):975-981
A strain of Erwinia aroideae produced an extracellular pectolytic enzyme under growth conditions with pectin or pectic acid as the inducer. This strain also produced a pectin lyase when nalidixic acid is added to a culture medium. The pectolytic enzyme produced under the growth conditions was purified approximately 40-fold from the culture fluid by carboxy- methyl cellulose and Sephadex G-75 gel column chromatographies. The purified enzyme was almost homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis, having a molecular weight of about 36,000 to 38,000. This enzyme, with optimal activity at pH 9.0 to 9.2, produced reaction products which had a strong absorption at 230 nm indicating a lyase type of the reaction. The enzyme activity was markedly stimulated by calcium ion and completely inhibited by cobalt and mercuric ions and by ethylenediaminetetraacetate. Pectic acid or pectin with lower methoxyl content was a good substrate for this enzyme, while no significant activity was observed when pectin with higher methoxyl content was used as a substrate. It was concluded that the enzyme produced under the normal growth conditions is an endo-pectate lyase and differs from the pectin lyase induced by nalidixic acid. 相似文献
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Shuwsei Kamimiya Tsuguaki Nishiya Kazuo Izaki Hajime Takahashi 《Bioscience, biotechnology, and biochemistry》2013,77(5):1071-1078
A strain of Erwinia aroideae produces a remarkable amount of pectolytic enzyme when the organism was induced by nalidixic acid for the bacteriocin production. This pectolytic enzyme was purified approximately 60-fold from the induced medium by carboxymethyl-cellulose and Sephadex G–75 gel column chromatographies after batchwise treatment with carboxymethyl- and diethylaminoethyl-celluloses. The purified enzyme was almost homogeneous on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, a molecular weight of about 28,000 to 32,000 was determined for this enzyme. The optimum pH of the enzyme activity was about 8.0 to 8.2. The purified enzyme produced reaction products from pectin and methoxylated pectic acid which had a strong absorption at 235 nm indicating a trans-eliminase reaction. Pectin or pectic acid with higher methoxyl content was a good substrate for this enzyme, while no significant activity was observed when pectic acid was a substrate. The limit of degradation of pectin and pectic acid with higher methoxyl content (90% esterified) by the enzyme were 6.5% and 43%, respectively. It was concluded that the enzyme is a new endo-pectin trans-eliminase from bacterial origin. 相似文献
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Shuwsei Kamimiya Kazuo Izaki Hajime Takahashi 《Bioscience, biotechnology, and biochemistry》2013,77(13):2367-2372
Pectolytic enzyme formation by whole cells of Erwinia aroideae was markedly stimulated when nalidixic acid was added to a culture medium. The activity of pectolytic enzyme was markedly stimulated by nalidixic acid when the activity was measured by the decrease of viscosity of pectin, while activities of both polygalacturonic acid trans-eliminase and polygalacturonase which were measured respectively by the increase of optical density at 230 nm and the liberation of aldehyde groups, were not stimulated. The analysis of pectolytic enzyme by carboxymethyl cellulose column chromatography indicated that there was a significant difference in the elution profiles between the pectolytic enzyme induced by nalidixic acid and that synthesized under normal conditions. Therefore, we conclude that two enzymes are distinct protein species. 相似文献
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Shuwsei Kamimiya Kazuo Izaki Hajime Takahashi 《Bioscience, biotechnology, and biochemistry》2013,77(5):911-912
Diploid baker’s yeast capable of strongly activating a mouse macrophage was constructed based on haploid mutant AQ-37 obtained previously. The obtained strain BQ-55 activated also human immune cells. To clarify a factor for the activation, the cell wall structure, especially the β-glucan structure, was analyzed, suggesting that the length of branching, β-1,6-glucan, may be one of the factors. 相似文献
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Hisashi Kamimiya Yusuke Suzuki Takeshi Kasama Hitomi Kajiwara Takeshi Yamamoto Toshiki Mine Shinobu Watarai Kiyoshi Ogura Kazuo Nakamura Junichi Tsuge Yasunori Kushi 《Journal of lipid research》2013,54(3):571-580
On the basis of the results outlined in our previous report, bacterial
sialyltransferases (ST) from marine sources were further characterized using
glycosphingolipids (GSL), especially ganglio-series GSLs, based on the enzymatic
characteristics and kinetic parameters obtained by Line weaver-Burk plots. Among
them, GA1 and GA2 were found to be good substrates for these unique STs. Thus,
new gangliosides synthesized by α2-3 and α2-6STs were structurally
characterized by several analytical procedures. The ganglioside generated by the
catalytic activity of α2-3ST was identified as GM1b. On the other hand,
when enzyme reactions by α2-6STs were performed using substrates GA2 and
GA1, very unique gangliosides were generated. The structures were identified as
NeuAcα2-6GalNAcβ1-4Galβ1-4Glcβ-Cer and
NeuAcα2-6Galβ1-3GalNAcβ1-4Galβ1-4Glcβ-Cer,
respectively. The synthesized ganglioside
NeuAcα2-6GalNAcβ1-4Galβ1-4Glcβ-Cer showed binding activity
to the influenza A virus {A/Panama/2007/99 (H3N2)} at a similar level to
purified sialyl(α2-3)paragloboside (S2-3PG) and
sialyl(α2-6)paragloboside (S2-6PG) from mammalian sources. The evidence
suggests that these STs have unique features, including substrate specificities
restricted not only to lacto-series but also to ganglio-series GSLs, as well as
catalytic potentials for ganglioside synthesis. This evidence demonstrates that
effective in vitro ganglioside synthesis could be a valuable tool for
selectively synthesizing sialic acid (Sia) modifications, thereby preparing
large-scale gangliosides and permitting the exploration of unknown
functions. 相似文献
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Tsuge J Hiratsuka H Kamimiya H Nozaki H Kushi Y 《Bioscience, biotechnology, and biochemistry》2008,72(10):2667-2674
Seven strains of fungi were isolated from activated sludge and identified as Mucor sp., Geotrichum sp., Trichosporon sp., Candida sp., and Trichoderma sp. by 28S rDNA D2 region sequences analysis. The structures of the main ceramide monosaccharides (CMSs) from these fungi were identified as glucosylceramide (GlcCer) consisting of ceramide moieties of 9-methyl-octadeca-sphingadienine (9-Me d18:2), with 2-hydroxyhexadecanoate (h16:0) (Mucor sp. and Geotrichum sp.), 2-hydroxyoctadecanoate (h18:0) (Trichosporon sp. and Candida sp.), and 2-hydroxyoctadecenoate (h18:1) (Trichoderma sp.). Seasonal changes in glycosphingolipids in activated sludge suggest the possibility that microbial flora in activated sludge changes with the seasons, and that fungi adaptable to low temperatures dominate in the cold period, resulting in the maintenance of stable effluent quality. Mucor sp., Geotrichum sp., and Candida sp. satisfactorily reduced the BOD of synthetic sewage at 10 degrees C. These results indicate that fungi in activated sludge can contribute to wastewater treatment in cold conditions. 相似文献
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Hirosato Tanaka Nagahiro Ogasawara Shuwsei Kamimiya 《Bioscience, biotechnology, and biochemistry》2013,77(7):1541-1552
Protoplasts of Pyricularia oryzae P2, a rice blast mold, were prepared in high yield from the young mycelium of the fungus using lytic enzymes from Bacillus circulans WL 12. The majority of the protoplasts had one nucleus per cell. The protoplasts formed a cell wall and eventually reverted to normal mycelial form in liquid medium. The process of regeneration was studied under phase-contrast and electron microscopes. The protoplast built a very thick wall prior to the protrusion of a germ-tube like hypha. Golgi apparatus-like structures appeared in the early stage of regeneration and disappeared later. Electron-transparent amorphous structures accumulated during regeneration. Lomasomes were observed in the regenerated cell walls. 相似文献
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