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101.
In this communication, we describe a novel and facile method for the immobilization of NAD(+)/NADH on an electrode surface using a hydrophobic ionic liquid, 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide ([C4mim][Tf(2)N]). By taking advantage of the insolubility of NAD(+)/NADH in hydrophobic ionic liquids, it is expected that NAD(+)/NADH can be retained on the electrode's surface. Alcohol dehydrogenase (ADH) and NAD(+)/NADH were immobilized with a gelatin hydrogel on an electrode that was modified with an electropolymerized ruthenium complex containing 5-amino-1,10-phenanthroline (pAPRu) as a mediator for NADH oxidation. The (ADH, NAD(+))/pAPRu-immobilized electrode exhibited the electrocatalytic oxidation of ethanol in [C4mim][Tf(2)N]. The obtained catalytic current in [C4mim][Tf(2)N] was comparable to that in buffer solution containing NAD(+). It was confirmed by UV-vis spectroscopy that NAD(+) did not dissolve in the [C4mim][Tf(2)N] and was retained on the electrode's surface. Furthermore, we succeeded in constructing an ethanol/O(2) biofuel cell comprised of an (ADH, NAD(+))/pAPRu anode and a bilirubin oxidase cathode using [C4mim][Tf(2)N] as an electrolyte. 相似文献
102.
Jiro Arima Yoshitaka Isoda Tadashi Hatanaka Nobuhiro Mori 《World journal of microbiology & biotechnology》2013,29(5):899-906
N-Acyl-d-amino acid amidohydrolases (d-aminoacylases) are often used as tools for the optical resolution of d-amino acids, which are important products with applications in industries related to medicine and cosmetics. For this study, genes encoding d-aminoacylase were cloned from the genomes of Streptomyces spp. using sequence-based screening. They were expressed by Escherichia coli and Streptomyces lividans. Almost all of the cell-free extracts exhibit hydrolytic activity toward N-acetyl-(Ac-)d-Phe (0.05–6.32 μmol min?1 mg?1) under conditions without CoCl2. Addition of 1 mM CoCl2 enhanced their activity. Among them, the highest activity was observed from cell-free extracts prepared from S. lividans that possess the d-aminoacylase gene of Streptomyces sp. 64E6 (specific activities were, respectively, 7.34 and 9.31 μmol min?1 mg?1 for N-Ac-d-Phe and N-Ac-d-Met hydrolysis). Furthermore, when using glycerol as a carbon source for cultivation, the recombinant enzyme from Streptomyces sp. 64E6 was produced in 4.2-fold greater quantities by S. lividans than when using glucose. d-Aminoacylase from Streptomyces sp. 64E6 showed optimum at pH 8.0–9.0. It was stable at pH 5.5–9.0 up to 30 °C. The enzyme hydrolyzed various N-acetyl-d-amino acids that have hydrophobic side chains. In addition, the activity toward N-chloroacetyl-d-Phe was 2.1-fold higher than that toward N-Ac-d-Phe, indicating that the structure of N-acylated portion of substrate altered the activity. 相似文献
103.
Akira Yanai Keijiro Kato Teruhiko Beppu Kei Arima 《Bioscience, biotechnology, and biochemistry》2013,77(8):1505-1508
A practical method for preparing peptidoglycan from Ps. aeruginosa and E. coli was devised. After bacterial cells were dissolved in boiling 4% SDS solution, peptidoglycan was collected and washed with water by centrifugation. Peptidoglycan was treated further with pronase and lyophilized. The final preparation of peptidoglycan from Ps. aeruginosa appeared as a filmy coagulation in electron micrograph and its amino acid composition was determined as follows: Glu/Ala/A2pm/Mur/GlcN (100/183/104/61/98). The lysozyme digest showed the same pattern as that of E. coli peptidoglycan. N-Terminal analysis suggested that about half of the peptide chains was interbridged by the peptide bond between Ala and A2pm. The probable ratio of muropeptides in the peptidoglycan was estimated. 相似文献
104.
Wen-Hsiung Liu Teruhiko Beppu Kei Arima 《Bioscience, biotechnology, and biochemistry》2013,77(11):2487-2492
The hydrolysis of olive oil by the Humicola lipase was inhibited by the addition of n-alcohols, fatty acids and surface active agents. The inhibition of n-alcohols was overcomed by the addition of more substrate but not by the addition of more enzyme. The inhibition of fatty acids and bile salts was eliminated by adding calcium ion. It was concluded that the inhibition of the Humicola lipase by n-alcohols, fatty acids and bile salts was not due to inactivation of the enzyme directly but due to the displacing of the substrate from the oil/water interface, thus blocking the enzyme from the substrate. 相似文献
105.
Kenji Mori Masanao Matsui Yusuke Sumiki 《Bioscience, biotechnology, and biochemistry》2013,77(3):205-222
In order to clarify the structure of ring A of gibberellins, thirteen lactones of cyclohexan series, of which eight were new, were prepared to examine their infrared spectra. So far; the experiment is concerned, γ-lactones show the characteristic absorption band in the rang 1775~1782 cm?1 in dioxane, while 5-ones in the range 1730/~1762 cm?1. Since the absorptio band due to lactone carbonyl of gibberellins occurs at the range 1777~1786 cm?1 in dioxane, the lactone ring of gibberellins seems to be γ. 相似文献
106.
Yuzuru Matsuda Teruhiko Beppu Kei Arima 《Bioscience, biotechnology, and biochemistry》2013,77(6):1179-1186
The Oxygen activating mechanism of Fusarium lipoxygenase, a heme-containing dioxygenase, was studied. The enzyme did not require any cofactors, such as H2O2, however, both superoxide dismutase and catalase inhibited linoleate peroxidation by Fusarium lipoxygenase. A low concentration of H2O2 caused a distinct acceleration in enzymatic peroxidation. These results indicate that both O2? and H2O2 are produced as essential intermediates of oxygen activation during formation of linoleate hydroperoxides by Fusarium lipoxygenase. This peroxidation reaction was also prevented by scavengers of singlet oxygen (1O2), but not by scavengers of hydroxy 1 radical (OH). Generation of O2? in the enzyme reaction was detected by its ability to oxidize epinephrine to adrenochrome. Moreover, the rate of peroxide formation was greater in the D2O than in the H2O buffer system. These results suggest that the Haber–Weiss reaction (O2?+H2O2→OH?+OH·+1O2) is taking part in linoleate peroxidation by Fusarium lipoxygenase, and the 1O2 evolved could be responsible for the peroxidation of linoleate. H2O2 produced endogenously in the enzyme reaction might act as an activating factor for the enzyme. This possible mechanism of oxygen activation can explain the absence of a need for exogenous cofactors with Fusarium lipoxygenase in contrast to an other heme-containing dioxygenase, tryptophan pyrrolase, which requires an exogenous activating factor, such as H2O2. 相似文献
107.
Kei Arima Teiiti Narasaki Yoshio Nakamura Gakuzo Tamura 《Bioscience, biotechnology, and biochemistry》2013,77(8):924-929
About 2000 strains of microorganisms were examined for lipoprotein lipase producibilities and some microorganisms were found to produce lipases similar to animal lipoprotein lipases.Microorganisms were cultured on solid media containing a serum-activated olive oil emulsion, and strains which formed a clear zone around the colony were collected. The collected microorganisms were cultured on liquid media containing 0.5% of olive oil by shaking and the culture filtrates were tested for lipoprotein lipase activity by a turbidity method. The superior lipoprotein lipase producers obtained belonged to genera of Serratia, Pseudomonas, Mucor, and Streptomyces. 相似文献
108.
Kei Arima Hisayoshi Okazaki Hiroko Ono Kiyotaka Yamada Teruhiko Beppu 《Bioscience, biotechnology, and biochemistry》2013,77(10):2313-2317
A mutant of Streptomyces fradiae which requires oleic acid for neomycin formation was isolated and the effects of exogenous fatty acids and other additives on the formation of neomycin were studied. Palmitic acid and high concentration of sodium ions could replace oleic acid in neomycin formation. The fatty acid spectrum of the mutant strain ST–5B was quite different from that of the parent strain 3123. The major fatty acid components of the mutant and the parent were anteiso 15:0 and iso 16: 0, respectively. However the fatty acid composition of the mutant was changed from the anteiso 15: 0-type to the parental iso 16: 0-type by the supplement of oleic acid or high concentration of sodium ions in the medium. In the case of palmitic acid, the major fatty acid component of the mutant cells was changed from anteriso 15: 0 to normal 16:0. The role of these additives in neomycin formation by the mutant is discussed. 相似文献
109.
Saburo Tamura Makoto Takai Satoshi Miyamoto Seiroku Sakai Masayoshi Goda Yusuke Sumiki 《Bioscience, biotechnology, and biochemistry》2013,77(10):773-777
Various O,O-dialkyl O-cyanophenyl phosphates and phosphorothioates were prepared and their biological activities were examined. Among them, O,O-dimethyl O- (4-chloro-2-cyanophenyl) phosphorothioate was found to have selective and high toxicity to houseflies. O,O-Dimethyl O- (4-cyanophenyl) phosphorothioate, O,O-diethyl O- (4-cyanophenyl) phosphorothioate and O,O-diethyl O- (2-chloro-4-cyanophenyl) phosphorothioate showed high insecticidal activty to American cockroaches, though the former two were not so effective to houseflies. The dimethyl esters of these series exhibited markedly lowered mammalian toxicity. Among the O-ethyl O-cyanophenyl phenylphosphonothioates, O-ethyl O- (2-chloro-4-cyanophenyl) phenylphosphonothioate was highly effective to mites, while less effective to insects. 相似文献
110.