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951.
Yoshikazu Takagi Takane Fujimori Tsuyoshi Hata Hajime Kaneko Kunio Kato 《Bioscience, biotechnology, and biochemistry》2013,77(3):705-706
Glutamine production was investigated by coupling of glutamine synthetase from Gluconobacter suboxydans with a sugar fermentation system of baker's yeast (energy generating system). Under the optimum condition, 22 mM glutamine was formed in 3 hr, and the yield was 92% based on the substrate glutamate. The first step of the process was the accumulation of fructose 1,6-diphosphate (FDP) as a reservoir of fermentation energy, in the presence of a high concentration of inorganic phosphate; and the second step was accomplished by coupling the degradation of FDP with glutamine synthetase reaction through an ADP-ATP system. The effects of enzyme concentration, additives in the reaction mixture and others on glutamine formation were investigated, and the importance of three factors was pointed out: (a) the ratio of activity of energy generating system to utilizing system, (b) contaminated enzyme(s) in the energy utilizing system and (c) the enzymatic properties of the energy utilizing system. 相似文献
952.
Tsuyoshi Nishitoba Hiroji Sato Takanori Kasai Hirokazu Kawagishi Sadao Sakamura 《Bioscience, biotechnology, and biochemistry》2013,77(6):1793-1798
Four new bitter terpenoids, lucidenic acids A (1), B (2), C (3) and ganoderic acid C (5), were isolated from the fruiting bodies of Ganoderma lucidum, together with the known bitter ganoderic acid B (4). On the basis of spectroscopic data and chemical conversion, their structures were determined to be 7β-hydroxy-4,4,14α-trimethyl-3,11,15-trioxo-5α-chol-8-en-24-oic acid, 7β,12β-dihydroxy-4,4,14α-trimethyl-3,11,15-trioxo-5α-chol-8-en-24-oic acid, 3β,7β,12β-trihydroxy-4,4,14α-trimethyl-11,15-dioxo-5α-chol-8-en-24-oic acid and 7β-hydroxy-3,11,15,23-tetraoxo-5α-lanost- 8-en-26-oic acid, respectively. 相似文献
953.
Tatsuo Tano Tsuyoshi Sugio Kazutami Imai 《Bioscience, biotechnology, and biochemistry》2013,77(3):695-696
High phytin containing particles were isolated from rice bran by a combination of a differetial centrifugation and an aqueous two phase system, using dextran 500 and polyethylene glycol 6000. The isolated particles consisted mainly of phytic acid, potassium and magnesium. The chemical composition and electron microscopic observation of the isolated particles confirmed that the electron dense material, which is embedded in aleurone particles of matured rice grains, is potassium and magnesium salts of phytic acid. 相似文献
954.
Takayuki Yamamoto Haruya Takahashi Koichiro Suzuki Akira Hirano Masanori Kamei Tsuyoshi Goto 《Bioscience, biotechnology, and biochemistry》2013,77(12):2059-2063
Several concentrations of theobromine (TB) and (?)-epicatechin (EC) were coadministered to rats, and plasma EC and its metabolites were determined using ultra-high-performance liquid chromatography–tandem mass spectrometry. It has been demonstrated that TB increases the absorption of EC in a dose-dependent manner. Cocoa powder had a similar effect, and the mechanism involved is not thought to depend on tight junctions. 相似文献
955.
The first step in branched-chain amino acid biosynthesis is catalyzed by acetohydroxyacid synthase (EC 2.2.1.6). This reaction involves decarboxylation of pyruvate followed by condensation with either an additional pyruvate molecule or with 2-oxobutyrate. The enzyme requires three cofactors, thiamine diphosphate (ThDP), a divalent ion, and flavin adenine dinucleotide (FAD). Escherichia coli contains three active isoenzymes, and acetohydroxyacid synthase I (AHAS I) large subunit is encoded by the ilvB gene. In this study, the ilvB gene from E. coli K-12 was cloned into expression vector pETDuet-1, and was expressed in E. coli BL21 (DH3). The purified protein was identified on a 12% SDS–PAGE gel as a single band with a mass of 65 kDa. The optimum temperature, buffer, and pH for E. coli K-12 AHAS I were 37 °C, potassium phosphate buffer, and 7.5. Km values for E. coli K-12 AHAS I binding to pyruvate, Mg+2, ThDP, and FAD were 4.15, 1.26, 0.2 mM, and 0.61 μM respectively. Inhibition of purified AHAS I protein was determined with herbicides and new compounds. 相似文献
956.
Tsuyoshi Sugio Kimihito Wada Wataru Mizunashi Kazutami Imai Tatsuo Tano 《Bioscience, biotechnology, and biochemistry》2013,77(11):2917-2918
The cDNA sequence coding for tuna growth hormone (tGH) was placed under the control of the repressible acid phosphatase (PHO5) promoter of a yeast, Saccharomyces cerevisiae, in an expression plasmid, pAM82. The yeast cells transformed with the plasmid synthesized tGH only when the cDNA was attached to the vector through a synthetic oligonucleotide linker having a similar sequence to the 5′-flanking region of the PHO5 structural region. The amount of tGH produced in yeast cells accounted for more than 3% of the total cellular protein and the product was immunologically identified as tGH by Western blotting using polyclonal antibodies specific to tGH. 相似文献
957.
958.
Masao Horiba Masayuki Nagase Yoshifumi Oki Seiya Yamamoto 《Bioscience, biotechnology, and biochemistry》2013,77(4):1095-1096
Monoclonal antibodies (MoAbs) reacting with human G-CSF and /or its muteins were established by cell fusion between P3.X63/Ag8.U1 myeloma cells and spleen cells from BALB/C mice immunized with recombinant human intact G-CSF or its mutein, designated ND28. Two MoAbs reacted with intact G-CSF and all kinds of muteins tested, designated KM341 and KM342, and two MoAbs specific for intact G-CSF, designated KM340 and KM343, were obtained from the mice immunized with recombinant human intact G-CSF.The sera from the mice immunized with a mutein of G-CSF, ND28, reacted with intact G-CSF and all muteins tested. Two MoAbs specific for ND28, designated KM498 and KM511, were obtained from these mice. These MoAbs seem to recognize the sequence of a few amino acids that is peculiar for ND28. However, the epitopes recognized by KM498 and KM511 were maybe subtly different, because KM498 and KM511 could not completely inhibit each other.Human G-CSF and/or its muteins could be measured by sandwich ELISA using these MoAbs with suitable combinations. The immuno-affinity column using KM342 or KM498 adsorbed G-CSFs or specifically ND28, previously. By elution with 0.15m NH4OH, the G-CSFs or ND28 were eluted with a high recovery. 相似文献
959.
Jiro Arima Shota Tokai Masanori Chiba Tsuyoshi Ichiyanagi Yukinori Yabuta Nobuhiro Mori 《Bioscience, biotechnology, and biochemistry》2013,77(11):1856-1863
Pleurotus eryngii serine aminopeptidase that has peptide bond formation activity, redesignated as eryngase, was cloned and expressed. Eryngase has a family S9 peptidase unit in the C-terminal region having a catalytic triad of Ser, Asp, and His. In the phylogenetic relations among the subfamilies of family S9 peptidase (S9A, prolyl oligopeptidase; S9B, dipeptidyl peptidase; S9C, acylaminoacyl peptidase; S9D, glutamyl endopeptidase), eryngase existed alone in the neighbor of S9C subfamily. Mutation of the active site Ser524 of the eryngase with Ala eliminated its catalytic activity. In contrast, S524C mutant maintained low catalytic activity. Investigation of aminolysis activity using l-Phe-NH2 as a substrate showed that S524C mutant exhibited no hydrolysis reaction but synthesized a small amount of l-Phe-l-Phe-NH2 by the catalysis of aminolysis. In contrast, wild-type eryngase hydrolyzed the product of aminolysis l-Phe-l-Phe-NH2. Results show that the S524C mutant preferentially catalyzed aminolysis when on an l-Phe-NH2 substrate. 相似文献
960.
Purification studies were conducted on DNA polymerase bound to the membrane fraction of E. coli HF 4704. Purified enzyme (Fraction V) required Mg2+ and showed an optimun pH of 7.2. Various kinds of salt indicated a stimulative effect at concentrations lower than 0.1 m. Fraction V was unstable at an acidic condition (pH 5.0) but was rather stable at an alkaline condition (pH 9.0). The enzyme activity was lost by incubation at 45°C for 30min but was stabilized by the addition of DNA. The enzyme contained exonuclease activity but no endonuclease activity. The enzyme produced only light density DNA of various sizes. The function of this enzyme as considered to fill single stranded region of the double stranded primer DNA. 相似文献