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81.
An α-glucosidase was purified from flint corn by precipitation with ammonium sulfate, chromatographies on CM-cellulose and Hydroxylapatite and gel-filtrations on Sephadex G-100. The purified enzyme was homogeneous in ultracentrifugal and disc electrophoretic analysis. The sedimentation coefficient was calculated to be 6.5 S. The molecular weight was estimated to be approximately 6.5×104 by gel-filtration technique.The optimal pH was found to be 3.6 for both maltose and soluble starch. The enzyme lost about 80% of the activity by incubation at 60°C for 10 min.The ratio of velocity of hydrolysis for maltose, phenyl-α-glucoside and soluble starch was estimated to be 100:14.3:6.1 in this order. The αglucosidase hydrolyzed soluble starch exo-wisely. 相似文献
82.
Yasuo Kitagawa Etsuro Sugimoto Hideo Chiba 《Bioscience, biotechnology, and biochemistry》2013,77(1):199-206
The amino acid composition of beef liver d-glycerate dehydrogenase (EC 1.1.1.29) was determined. Results of sodium dodecyl sulfate gel electrophoresis and measurements of the number of NADH bound by the enzyme and the number of the essential sulfhydryl groups suggested that the enzyme was composed of two identical subunits with the molecular weight of 36,000. Close relation between the essential sulfhydryl groups and the coenzyme binding site was also suggested. Effect of an alkylating agent (bromopyruvate) with the structure similar to the substrate was studied. Effects of iodoacetate and iodoacetamide were also studied. It was suggested that these reagents behaved as active-site-directed irreversible inhibitors of the enzyme. Bromopyruvate exhibited a high affinity to the enzyme. Iodoacetate (anionic reagent) had a higher affinity than iodoacetamide (neutral reagent). 相似文献
83.
Hirokazu Matsui Seiya Chiba Tokuji Shimomura Nobuyuki Takahashi 《Bioscience, biotechnology, and biochemistry》2013,77(11):2239-2240
Hen lysozyme modified with histamine (HML) and Japanese quail lysozyme (JQL) were treated with immobilized metal ion affinity chromatography to analyze the states of their imidazole groups. When Ni(II) was used as the metal ion immobilized, JQL was strongly retained in a Ni(II)-chelating Sepharose column, while hen lysozyme and HML were hardly retained in the same column. All of these lysozymes have a histidine imidazole group at the 15th position, while JQL has an additional histidine imidazole group at the 103rd position and HML has an additional imidazole group covalently attached to Asp101. Thus, I concluded that the imidazole group at the 103rd position of JQL is exposed to the solvent and recognized by the metal ion, but that the imidazole group attached to Asp101 in HML is localized to a hydrophobic region and not recognized by the metal ion. 相似文献
84.
Takeshi Saeki Seiya Chiba Tokuji Shimomura 《Bioscience, biotechnology, and biochemistry》2013,77(2):551-552
A water-soluble and neutral polysaccharide was extracted from the current pseudobulbs of Oncidium “Gower Ramsey” during the early inflorescence stage (flower stalk less than 4 cm) by hot water, precipitated with ethanol, and purified with an anion exchanger. From the data of monosaccharide composition and linkage and anomeric configuration analyses, the polysaccharide was identified as a linear β-1→4 linked mannan. 相似文献
85.
Etsuro Sugimoto Masaaki Yoshikawa Yutaka Nishida Hideo Chiba 《Bioscience, biotechnology, and biochemistry》2013,77(12):2497-2504
It was indicated from fluorescence spectra and fluorescence titration that a hydrophobic probe, 1-anilino-8-naphthalenesulfonate (ANS), binds to casein components (αs-, β- and κ-caseins). Fluorescence intensity and affinity of ANS-κ-casein complex were larger than that of ANS-αs- and ANS-β-casein complexes. Enhancements of fluorescence intensity of complexes of casein components were observed by the addition of KCI or CaCl2. Reason for the enhancement was postulated to be the increase of the quantum yield of the ANS fluorescence caused by the environmental change of ANS binding region of the casein components.Marked increase of sedimentation coefficient of β-casein in the presence of KCl or CaCl2 at 10°C was caused by the addition of ANS. This may be responsible for the stimulation of the Ca-dependent precipitation of β-casein by the addition of ANS.It was found that αs · κ-association was prevented by ANS and that hydrophobic interaction have an important role for αs · κ-association. 相似文献
86.
Koji Ikura Hiroshi Sakurai Katsuzumi Okumura Ryuzo Sasaki Hideo Chiba 《Bioscience, biotechnology, and biochemistry》2013,77(12):3527-3531
Guinea pig liver transglutaminase was purified in a yield of more than 80% by a one-step purification procedure using an immunoadsorbent column of monoclonal antibodies. Active enzyme could be recovered by alkaline buffer desorption and quick neutralization. The purified enzyme was more than 98% homogeneous on Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, and had the same enzymological and immunological properties as those of the enzyme purified by conventional procedures. 相似文献
87.
Masaaki Yoshikawa Etsuro Sugimoto Hideo Chiba 《Bioscience, biotechnology, and biochemistry》2013,77(5):1005-1013
It was indicated from ultraviolet difference spectra and ultracentrifugal experiments that associations occurred between two casein components (αs- and κ-caseins, β- and κ-caseins and αs- and β-caseins) at lower CaCl2 concentrations (2~3 mm) and that aromatic amino acid residues participated in the associations. Chemical modification studies with 2-hydroxy-5-nitrobenzylbromide indicated that tryptophane residues of each casein component were not essential for these associations. It was also demonstrated by nitration of tyrosine residues with tetranitromethane that tyrosine residues of κ-casein were essential for αs·κ-association and for β·κ-association and that tyrosine residues of αs-casein were important to αs·β-association.Interactions between casein components were also studied at higher CaCl2 concentration (10 mm) which is enough for micelle formation. It was found that tyrosine residues of κ- casein played an important role for the stabilization of αs- and β-caseins. Properties of the nitrated-β-casein were almost the same as that of the native β-casein except the absorption spectrum. αs·β-Interaction in the presence of 10 mm CaCl2 was investigated by use of the nitrated-β-casein instead of the native β-casein. It was proved that αs-casein was stabilized by the nitrated-β-casein and that precipitation of the nitrated-β-casein increased in the presence of αs-casein.The mechanism of interactions between casein components at higher CaCl2 concentration (10 mm) are discussed in connection with the associations at lower CaCl2 concentrations (2~3 mm). 相似文献
88.
Kazunori Sasaki Tomoyuki Oki Yumi Kai Yoichi Nishiba 《Bioscience, biotechnology, and biochemistry》2013,77(8):1308-1314
The purpose of this study was to investigate the effect of repeated harvesting on the content of caffeic acid (CA) and seven species of caffeoylquinic acids (CQAs) in sweet potato leaves using a newly developed high-performance liquid chromatography method. Six cultivars and two breeding lines were used in this study. Leaves were collected at monthly intervals from 1st harvest (May) to 4th harvest (August) in 2011 and 2012. ANOVA analysis revealed that the contents of CQAs were significantly different among all cultivars and breeding lines, but no significant differences were found for CA. No annual variation was confirmed in CA and CQAs. Repeated harvest of sweet potato leaves affected the content of only 4-CQA and 5-CQA. Post-hoc comparisons using Tukey’s method indicated that the contents of 4-CQA and 5-CQA in sweet potato leaves harvested at first time were significantly higher compared to those at the other harvest times. 相似文献
89.
Shoji Hayashi Alexandra Houssaye Yasuhisa Nakajima Kentaro Chiba Tatsuro Ando Hiroshi Sawamura Norihisa Inuzuka Naotomo Kaneko Tomohiro Osaki 《PloS one》2013,8(4)
Background
The paleoecology of desmostylians has been discussed controversially with a general consensus that desmostylians were aquatic or semi-aquatic to some extent. Bone microanatomy can be used as a powerful tool to infer habitat preference of extinct animals. However, bone microanatomical studies of desmostylians are extremely scarce.Methodology/Principal Findings
We analyzed the histology and microanatomy of several desmostylians using thin-sections and CT scans of ribs, humeri, femora and vertebrae. Comparisons with extant mammals allowed us to better understand the mode of life and evolutionary history of these taxa. Desmostylian ribs and long bones generally lack a medullary cavity. This trait has been interpreted as an aquatic adaptation among amniotes. Behemotops and Paleoparadoxia show osteosclerosis (i.e. increase in bone compactness), and Ashoroa pachyosteosclerosis (i.e. combined increase in bone volume and compactness). Conversely, Desmostylus differs from these desmostylians in displaying an osteoporotic-like pattern.Conclusions/Significance
In living taxa, bone mass increase provides hydrostatic buoyancy and body trim control suitable for poorly efficient swimmers, while wholly spongy bones are associated with hydrodynamic buoyancy control in active swimmers. Our study suggests that all desmostylians had achieved an essentially, if not exclusively, aquatic lifestyle. Behemotops, Paleoparadoxia and Ashoroa are interpreted as shallow water swimmers, either hovering slowly at a preferred depth, or walking on the bottom, and Desmostylus as a more active swimmer with a peculiar habitat and feeding strategy within Desmostylia. Therefore, desmostylians are, with cetaceans, the second mammal group showing a shift from bone mass increase to a spongy inner organization of bones in their evolutionary history. 相似文献90.
Yoshiki Koriyama Yusuke Takagi Kenzo Chiba Matsumi Yamazaki Kayo Sugitani Kunizo Arai Hirokazu Suzuki Satoru Kato 《PloS one》2013,8(8)
Like other CNS neurons, mature retinal ganglion cells (RGCs) are unable to regenerate their axons after nerve injury due to a diminished intrinsic regenerative capacity. One of the reasons why they lose the capacity for axon regeneration seems to be associated with a dramatic shift in RGCs’ program of gene expression by epigenetic modulation. We recently reported that (1R)-isoPropyloxygenipin (IPRG001), a genipin derivative, has both neuroprotective and neurite outgrowth activities in murine RGC-5 retinal precursor cells. These effects were both mediated by nitric oxide (NO)/S-nitrosylation signaling. Neuritogenic activity was mediated by S-nitrosylation of histone deacetylase-2 (HDAC2), which subsequently induced retinoic acid receptor β (RARβ) expression via chromatin remodeling in vitro. RARβ plays important roles of neural growth and differentiation in development. However, the role of RARβ expression during adult rat optic nerve regeneration is not clear. In the present study, we extended this hypothesis to examine optic nerve regeneration by IPRG001 in adult rat RGCs in vivo. We found a correlation between RARβ expression and neurite outgrowth with age in the developing rat retina. Moreover, we found that IPRG001 significantly induced RARβ expression in adult rat RGCs through the S-nitrosylation of HDAC2 processing mechanism. Concomitant with RARβ expression, adult rat RGCs displayed a regenerative capacity for optic axons in vivo by IPRG001 treatment. These neuritogenic effects of IPRG001 were specifically suppressed by siRNA for RARβ. Thus, the dual neuroprotective and neuritogenic actions of genipin via S-nitrosylation might offer a powerful therapeutic tool for the treatment of RGC degenerative disorders. 相似文献