共查询到20条相似文献,搜索用时 15 毫秒
1.
Chieko Kitada Masahiko Fujino Youji Sakagami Mitsuru Yoshida Akira Isogai Akinori Suzuki 《Bioscience, biotechnology, and biochemistry》2013,77(4):1049-1051
Myosin and reconstituted actomyosin free of regulatory proteins were mixed in 0.6 m KCl and 20 mm phosphate buffer at pH 6.0 and were tested quantitatively for thermally induced gelation properties by measuring the rigidity of the system at 65°C. Full enhancement of gelation was attained when the weight ratio of myosin-to-actomyosin was about 4. The addition of regulatory proteins to actomyosin could restore calcium sensitivity of the contractile system, but did not affect the heat-induced gelation of myosin in the presence of actomyosin, suggesting that regulatory proteins play no role in the heat-induced gelation of the system.Neither the single and double headed subfragments, both capable of interacting with F-actin, nor the helical tail subfragments, devoid of the intracting site with F-actin, exhibited changes in thermogelling properties when mixed with F-actin. However, upon addition to F-actomyosin, the tail subfragments revealed a significant effect on the gelation of actomyosin, whereas the headed subfragments exerted no influence over gelation of the system. These results indicate that the enhancing effect of F-actin on the heat-induced gelation of myosin was brought about solely by the limited amount of F-actomyosin formed in the system, which acts as a cross-linker between the tail portion of bound and free myosin molecules. 相似文献
2.
Regulatory properties of the enzymes in l-tyrosine and l-phenyalanine terminal pathway in Corynebacterium glutamicum were investigated. Prephenate dehydrogenase was partially feedback inhibited by l-tyrosine. Prephenate dehydratase was strongly inhibited by l-phenylalanine and l-tryptophan and 100% inhibition was attained at the concentrations of 5 × 10?2mm and 10?1mm, respectively. l-Tyrosine stimulated prephenate dehydratase activity (6-fold stimulation at 1 mm) and restored the enzyme activity inhibited by l-phenylalanine or l-tryptophan. These regulations seem to give the balanced synthesis of l-tyrosine and l-phenyl-alanine. Prephenate dehydratase from C. glutamicum was stimulated by l-methionine and l-leucine similarly to the enzyme in Bacillus subtilis and moreover by l-isoleucine and l-histidine. C. glutamicum mutant No. 66, an l-phenylalanine producer resistant to p-fluorophenyl-alanine, had a prephenate dehydratase completely resistant to the inhibition by l-phenylalanine and l-tryptophan. 相似文献
3.
Shin-Ya Tanimoto Michiko Yamashita Soichi Arai Masao Fujimaki 《Bioscience, biotechnology, and biochemistry》2013,77(6):1207-1210
The plastein formation by α-chymotrypsin from an ovalbumin hydrolysate was affected in an order of valency of salts when the concentration of each salt was 1 m. Monovalent cations were rather effective at this concentration and enhanced the plastein yield by 10%. In the presence of NaCl, the plastein formation showed two distinct maximal rates at its concentrations of 0.1 m and 0.8 m. The first maximum was considered to be resulted from an increase in enzyme activity, since chymotryptic hydrolysis of both N-acetyl-l-tyrosine ethyl ester and benzyloxycarbonyl-l-phenylalanine p-nitrophenyl ester was activated at an NaCl concentration of 0.1 ~ 0.2 m. The second maximum was ascribed to the salting-out of the product due to the higher concentration of NaCl. A salt-tolerant protease was also used to confirm the above conclusions. It was observed that this enzyme was much effective in producing a plastein at a high NaCl concentration. This may be due to the fact that both the enzyme activation effect and the product salting-out effect participate co-operatively. 相似文献
4.
Sakamura Sadao Chida Tsutomu Ito Jiro Sakai Ryutaro 《Bioscience, biotechnology, and biochemistry》2013,77(11):1810-1811
Bovine serum albumin was reduced by incubating with various concentrations (0–200 mM) of 2-mercaptoethanol, and its emulsifying properties were examined for an oil-in-water emulsion system. A particle size analysis revealed that albumin reduced at 30 mM of the thiol yielded smaller oil particles than either native protein, or the protein reduced at 70 or 200 mM of the thiol. Furthermore, the particle size was almost constant during 35 days of storage with albumin reduced at 30 mM of the thiol, while an emulsion prepared using the native protein, or the protein reduced at 70 or 200 mM of the thiol was unstable during the same storage period. Gel filtration chromatography and transmission electron micrography show that serum albumin made aggregates with high molecular size by its disulfide reduction with 70 or 200 mM, but not with 30 mM of 2-mercaptoethanol. It was, therefore, concluded that the emulsifying property of serum albumin can be improved by a mild disufide reduction. 相似文献
5.
Takanori Kasai Michiko Okuda Sadao Sakamura 《Bioscience, biotechnology, and biochemistry》2013,77(11):2723-2724
We found a potent platelet aggregation inhibitor in the culture broth of Streptomyces strain M-193. The inhibitor was purified as white crystals and investigated; it was identified as staurosporine. It strongly inhibited the platelet aggregation induced by collagen or ADP with IC50 values of 3.4 μm, and 11.6 μm, respectively. The inhibitor had no effect on thrombin-induced platelet aggregation or membrane stabilization against heat-induced hemolysis. 相似文献
6.
Hajime Hatta Naofumi Kitabatake Etsushiro Doi 《Bioscience, biotechnology, and biochemistry》2013,77(8):2083-2089
The turbidity and hardness of a heat-induced gel prepared from ovalbumin were examined at various pHs and ionic strengths. Depending on the conditions of the medium, a transparent solution, transparent gel, turbid gel, or turbid suspension was obtained by heating. The hardness was a maximum with the conditions that gave a transparent or slightly turbid gel. The gel and coagulums were solubilized by 1% SDS, but not by 6 m urea or 50 mm mercaptoethanol. The solution obtained by SDS treatment contained polymers shorter than octamers. 相似文献
7.
《Bioscience, biotechnology, and biochemistry》2013,77(10):1730-1735
L-Tartrate in wines and grapes was enzymatically quantified by using the secondary activity of D-malate dehydrogenase (D-MDH). NADH formed by the D-MDH reaction was monitored spectrophotometrically. Under the optimal conditions, L-tartrate (a 1.0 mM sample solution) was fully oxidized by D-MDH in 30 min. A linear relationship was obtained between the absorbance difference and the L-tartrate concentration in the range of a 0.02-1.0 mM sample solution with a correlation coefficient of 0.9991. The relative standard deviation from ten measurements was 1.71% at the 1.0 mM sample solution level. The proposed method was compared with HPLC, and the values determined by both methods were in good agreement. 相似文献
8.
Yoshiro Kamata Jun-ichi Kimigafukuro Kazuo Shibasaki 《Bioscience, biotechnology, and biochemistry》2013,77(9):1817-1823
Digestibilities of native, 5 m urea-denatured and 8 m urea-denatured glycinin were studied. Urea was removed by dialysis before digestion. The tryptic digestion of the proteins are influenced by ionic strength. Under low ionic strength condition (0 m NaCl), the proteins, even native glycinin, are well degraded. On the other hand, under high ionic strength condition (0.5 m NaCl), native glycinin resists the tryptic attack and 5 m urea-denatured glycinin is best degraded. The digestibility of 8 m urea-denatured glycinin is lower than that of 5 m urea-denatured one under the condition. The gel filtration and electrophoretic properties show that the digestion intermediate like glycinin-T (the intermediate from native glycinin) is contained in the digestion products. These suggest that the urea-denatured protein contains the almost renatured component after removal of urea. A larger amount of the glycinin-T-like protein was detected at 8 m urea denaturation than at 5 m urea. Therefore, glycinin renatures more readily from 8 m urea denaturation. Probably this is the cause of the decreased digestibility at 8 m urea denaturation. 相似文献
9.
Xylanase induction by β-xyloside was investigated in non-growing conditions using non-induced mycelia of Streptomyces sp. No. 3137 harvested from glucose medium. The mycelia started to produce xylanase without lag time when β-xyloside was added. The rate of xylanase synthesis was dependent on the concentration of β-xyloside added to the inducing culture medium. The induction constants of various β-xylosides were calculated from the Lineweaver-Burk plots; those of methyl-, isopropyl-, butyl- and ethylencyanohydrin-β-d-xylosides were 10.53 mm, 3.83 mm, 0.55mm and 0.25 mm, respectively. Some α-xylosides repressed xylanase synthesis. The rate of xylanase synthesis decreased suddenly after the addition of α-xyloside. The inhibition constants of methyl-, ethyl- and isopropyl-α-d-xylosides were 8.80 mm, 12.50 mm and 33.33 mm, respectively. The xylanase induction was also repressed by glucose. However, this repression was completely restored after consuming additional glucose. 相似文献
10.
Yoshiki Katoh Akira Kuninaka Hiroshi Yoshino 《Bioscience, biotechnology, and biochemistry》2013,77(10):1957-1962
Poly (A) took stable double-stranded form in 0.02 m phosphate buffer pH 5.8 when its S-value was higher than 3.5. Poly (C) also took stable double-stranded form in 0.01 m acetate buffer pH 3.9 containing 0.1 m NaCl if its S-value was higher than 4.8.Poly (I) with S-values higher than 6.5 stably formed triple-stranded form. Poly (A) with S-values over 3S and poly (I) with S-values over 7S were observed to aggregate in the presence of high concentration of salt, but poly (C) and poly (U) did not aggregate under the same condition. Effect of salt concentration differed with different polynucleotides and with different salts. 相似文献
11.
d-Arabinose(l-fucose) isomerase (d-arabinose ketol-isomerase, EC 5.3.1.3) was purified from the extracts of d-arabinose-grown cells of Aerobacter aerogenes, strain M-7 by the procedure of repeated fractional precipitation with polyethylene glycol 6000 and isolating the crystalline state. The crystalline enzyme was homogeneous in ultracentrifugal analysis and polyacrylamide gel electrophoresis. Sedimentation constant obtained was 15.4s and the molecular weight was estimated as being approximately 2.5 × 105 by gel filtration on Sephadex G-200.Optimum pH for isomerization of d-arabinose and of l-fucose was identical at pH 9.3, and the Michaelis constants were 51 mm for l-fucose and 160 mm for d-arabinose. Both of these activities decreased at the same rate with thermal inactivation at 45 and 50°C. All four pentitols inhibited two pentose isomerase activities competitively with same Ki values: 1.3–1.5 mm for d-arabitol, 2.2–2.7 mm for ribitol, 2.9–3.2 mm for l-arabitol, and 10–10.5 mm for xylitol. It is confirmed that the single enzyme is responsible for the isomerization of d-arabinose and l-fucose. 相似文献
12.
Production of d-xylose and l-arabinose isomerases by lactic acid bacteria was greatly promoted by the addition of manganese ions in cultural medium. Effective concentration of the ions was 5 × 1O-3 m. Ferrous ions were also effective for the production of d-xylose isomerase and cobaltous ions were somewhat effective for the production of l-arabinose isomerase. Zinc and cadmium ions inhibited bacterial growth. It was possible to increase the production of isomerase by changing MnSO4 concentration to 5× 10-3 m (0.l1 %) in place of 0.001 per cent in the normal medium.Column chromatographic procedures for the purification of pentose isomerases were carried out. Cation and anion exchange resins were not suitable because of their low exchange capacities and instability of the enzyme at acidic pH range. But the isomerases were successfully purified by DEAE-cellulose column chromatography with high recovery (85~90%). Using a Tris buffer, KCl concentration was increased in gradient. d-Xylose isomerase was eluted at pH 7.0 at 0~0.2 m KCl, and l-arabinose isomerase at pH 8.0 at 0~0.4 m KCl. The purified isomerases, d-xylose isomerase and l-arabinose isomerase, both required manganese ions specifically for their activities.D-Xylose isomerase and l-arabinose isomerase are different enzymes which can be separated from each other with acetone fractionation at pH 4.8~5.0, heat treatment or chromatography on a colnmn of DEAE-cellulose. In DEAE-cellulose chromatography with a linear gradient elution method, d-xylose isomerase is recovered in the first peak at pH 7.0 (Tris bnffer) with 0~0.2 m KCl, and l-arabinose isomerase is eluted in the second peak at pH 8.0 (Tris buffer) with a larger ionic strength. 相似文献
13.
Yuji Koide Mamoru Honma Tokuji Shimomura 《Bioscience, biotechnology, and biochemistry》2013,77(7):1171-1177
Branched chain amino acid aminotransferase was partially purified from Pseudomonas sp. by ammonium sulfate fractionation, aminohexyl-agarose and Bio-Gel A-0.5 m column chromatography.This enzyme showed different substrate specificity from those of other origins, namely lower reactivity for l-isoleucine and higher reactivity for l-methionine.Km values at pH 8.0 were calculated to be 0.3 mm for l-leucine, 0.3 mm for α-ketoglutarate, 1.1 mm for α-ketoisocaproate and 3.2 mm for l-glutamate.This enzyme was activated with β-mercaptoethanol, and this activated enzyme had different kinetic properties from unactivated enzyme, namely, Km values at pH 8.0 were calculated to be 1.2 mm for l-leucine, 0.3 mm for α-ketoglutarate.Isocaproic acid which is the substrate analog of l-leucine was competitive inhibitor for pyridoxal form of unactivated and activated enzymes, and inhibitor constants were estimated to be 6 mm and 14 mm, respectively. 相似文献
14.
Yoshio Kawai Yasuo Kawai Teijiro Uemura 《Bioscience, biotechnology, and biochemistry》2013,77(5):395-402
The present investigation is concerned with l-glutamic acid production in the presence of pyrrolidone carboxylic acid and glucose in Bacillus megaterium st. 6126. This strain does not grow on dl-pyrrolidone carboxylic acid (dl-PCA)1) as the sole source of carbon and nitrogen. The optimal concentration of yeast extract required for the maximal production of l-glutamic acid was 0.005% under the conditions used. As the yeast extract concentration was increased, growth increased proportionally; but the l-glutamic acid production did not exceed the control’s to which glucose and ammonium chloride had been added. l-Glutamic acid produced by both growing cultures and resting cells was derived from glucose and ammonium salt of dl-PCA. Isotope experiments suggested that the l-glutamic acid produced was partially derived from ammonium salt of dl-PCA in the growing culture which had been supplemented with d-glucose-U-14C or dl-PCA-1-14C and that ammonium salt of dl-PCA was consumed as the source of nitrogen and carbon for l-glutamic acid. 相似文献
15.
Regulatory properties of chorismate mutase from Corynebacterium glutamicum were studied using the dialyzed cell-free extract. The enzyme activity was strongly feedback inhibited by l-phenylalanine (90% inhibition at 0.1~1 mm) and almost completely by a pair of l-tyrosine and l-phenylalanine (each at 0.1~1 mm). The enzyme from phenylalanine auxotrophs was scarcely inhibited by l-tyrosine alone but the enzyme from a wild-type strain or a tyrosine auxotroph was weakly inhibited by l-tyrosine alone (40~50% inhibition, l-tyrosine at 1 mm). The enzyme activity was stimulated by l-tryptophan and the inhibition by l-phenylalanine alone or in the simultaneous presence of l-tyrosine was reversed by l-tryptophan. The Km value of the reaction for chorismate was 2.9 } 10?3 m. Formation of chorismate mutase was repressed by l-phenylalanine. A phenylalanine auxotrophic l-tyrosine producer, C. glutamicum 98–Tx–71, which is resistant to 3-amino-tyrosine, p-aminophenylanaine, p-fluorophenylalanine and tyrosine hydroxamate had chorismate mutase derepressed to two-fold level of the parent KY 10233. The enzyme in C. glutamicum seems to have two physiological roles; one is the control of the metabolic flow to l-phenylalanine and l-tyrosine biosynthesis and the other is the balanced partition of chorismate between l-phenylalanine-l-tyrosine biosynthesis and l-tryptophan biosynthesis. 相似文献
16.
《Bioscience, biotechnology, and biochemistry》2013,77(8):1652-1655
D-Galactosyl-β1→4-L-rhamnose (GalRha) was produced enzymatically from 1.1 M sucrose and 1.0 M L-rhamnose by the concomitant actions of four enzymes (sucrose phosphorylase, UDP-glucose-hexose 1-phosphate uridylyltransferase, UDP-glucose 4-epimerase, and D-galactosyl-β1→4-L-rhamnose phosphorylase) in the presence of 1.0 mM UDP-glucose and 30 mM inorganic phosphate. The accumulation of GalRha in 1 liter of the reaction mixture reached 230 g (the reaction yield was 71% from L-rhamnose). Sucrose and fructose in the reaction mixture were removed by yeast treatment, but isolation of GalRha by crystallization after yeast treatment was unsuccessful. Finally, 49 g of GalRha was isolated from part of the reaction mixture with yeast treatment by gel-filtration chromatography. 相似文献
17.
Homoserine dehydrogenases and aspartokinases in l-threonine- or l-threonine and l-lysine-producing mutants derived from Corynebacterium glutamicum KY 9159 (Met?) were studied with respect to the sensitivity to the inhibition by end products, l-threonine and l-lysine. The activities of homoserine dehydrogenases in the mutants which produced l-threonine or l-threonine and l-lysine were slightly less susceptible to the inhibition by l-threonine than the activity in the parent strain, KY 9159. The aspartokinases in the threonine-producing mutants, KY 10484 and KY 10230, which were resistant to α-amino-β-hydroxylvaleric acid (AHV, a threonine analog) and more sensitive to thialysine (a lysine analog) than the parent, were sensitive to the concerted feedback inhibition by l-lysine and l-threonine by about the same degree as KY 9159. The aspartokinase in an AHV- and thialysine-resistant mutant, KY 10440, which was derived from KY 10484 and produced about 14 mg/ml of l-threonine in a medium containing 10% glucose was less susceptible to the concerted feedback inhibition than KY 10484 or KY 9159, although the activity was still under the feedback control. In the parent strain, l-threonine activated aspartokinase activity in the absence of ammonium sulfate, an activator of the enzyme, but partially inhibited the activity in the presence of the salt. On the other hand, the enzyme of KY 10440 was activated by l-threonine either in the presence or in the absence of the salt. In another AHV- and thialysine-resistant mutant, KY 10251, which was derived from KY 10230 and produced both 9 mg/ml of l-threonine and 5/5 mg/ml of l-lysine, l-threonine and l-lysine simultaneously added hardly inhibited the activity of aspartokinase.Implications of these results are discussed in relation to l-threonine or l-lysine production, AHV or thialysine resistance and regulation of l-threonine biosynthesis in these mutants. 相似文献
18.
Gen-ichi Danno Sadahiko Yoshimura Masato Natake 《Bioscience, biotechnology, and biochemistry》2013,77(3):284-292
d-Glucose-isomerizing enzyme has been extracted in high yield from d-xylose-grown cells of Bacillus coagulans, strain HN-68, by treating with lysozyme, and purified approximately 60-fold by manganese sulfate treatment, fractionation with ammonium sulfate and chromatography on DEAE-Sephadex column. The purified d-glucose-isomerizing enzyme was homogeneous in polyacrylamide gel electrophoresis and ultracentrifugation and was free from d-glucose-6-phosphate isomerase. Optimum pH and temperature for activity were found to be pH 7.0 and 75°C, respectively. The enzyme required specifically Co++ with suitable concentration for maximal activity being 10?3 m. In the presence of Co++, enzyme activity was inhibited strongly by Cu++, Zn++, Ni++, Mn++ or Ca++. At reaction equilibrium, the ratio of d-fructose to d-glucose was approximately 1.0. The enzyme catalyzed the isomerization of d-glucose, d-xylose and d-ribose. Apparent Michaelis constants for d-glucose and d-xylose were 9×10?2 m and 7.7×10?2 m, respectively. 相似文献
19.
The crystalline d-mannitol dehyrogenase (d-mannitol:NAD oxidoreductase, EC 1.1.1.67) catalyzed the reversible reduction of d-fructose to d-mannitol. d-Sorbitol was oxidized only at the rate of 4% of the activity for d-mannitol. The enzyme was inactive for all of four pentitols and their corresponding 2-ketopentoses. The apparent optimal pH for the reduction of d-fructose or the oxidation of d-mannitol was 5.35 or 8.6, respectively. The Michaelis constants were 0.035 m for d-fructose and 0.020 m for d-mannitol. The enzyme was also found to be specific for NAD. The Michaelis constans were 1 × 10?5 m for NADH2 and 2.7 × 10?4 m for NAD. 相似文献
20.
The irreversibility of the dissociation of “myosin B” stored in 0.6 m KCl at pH 5.7 and 3°C was attributed to the rapid denaturation of F-actin dissociated from “myosin B”F-Actin was less stable than myosin A, in 0.18~0.60 m KCl at pH 5.7 and temperatures between 0 ~3°C.The decrease in the ability of F-actin to bind with myosin A was slightly dependent on storage temperature, and there was no apparent relation with the decrease in the solubility.A hypothetical scheme for F-actin denaturation was proposed. 相似文献