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101.
102.
Mechanism‐Based Trapping of the Quinonoid Intermediate by Using the K276R Mutant of PLP‐Dependent 3‐Aminobenzoate Synthase PctV in the Biosynthesis of Pactamycin
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Akane Hirayama Dr. Akimasa Miyanaga Prof. Dr. Fumitaka Kudo Prof. Dr. Tadashi Eguchi 《Chembiochem : a European journal of chemical biology》2015,16(17):2484-2490
Mutational analysis of the pyridoxal 5′‐phosphate (PLP)‐dependent enzyme PctV was carried out to elucidate the multi‐step reaction mechanism for the formation of 3‐aminobenzoate (3‐ABA) from 3‐dehydroshikimate (3‐DSA). Introduction of mutation K276R led to the accumulation of a quinonoid intermediate with an absorption maximum at 580 nm after the reaction of pyridoxamine 5′‐phosphate (PMP) with 3‐DSA. The chemical structure of this intermediate was supported by X‐ray crystallographic analysis of the complex formed between the K276R mutant and the quinonoid intermediate. These results clearly show that a quinonoid intermediate is involved in the formation of 3‐ABA. They also indicate that Lys276 (in the active site of PctV) plays multiple roles, including acid/base catalysis during the dehydration reaction of the quinonoid intermediate. 相似文献
103.
Shuyan Gao Naoto Koshizaki Hideo Tokuhisa Emiko Koyama Takeshi Sasaki Jae‐Kwan Kim Joonghyun Ryu Deok‐Soo Kim Yoshiki Shimizu 《Advanced functional materials》2010,20(1):78-86
Colloidal Au‐amplified surface plasmon resonance (SPR), like traditional SPR, is typically used to detect binding events on a thin noble metal film. The two major concerns in developing colloidal Au‐amplified SPR lie in 1) the instability, manifested as a change in morphology following immersion in organic solvents and aqueous solutions, and 2) the uncontrollable interparticle distance, determining probe spacing and inducing steric hindrance between neighboring probe molecules. This may introduce uncertainties into such detecting techniques, degrade the sensitivity, and become the barricade hampering colloidal Au‐based transducers from applications in sensing. In this paper, colloidal Au‐amplified SPR transducers are produced by using ultrathin Au/Al2O3 nanocomposite films via a radio frequency magnetron co‐sputtering method. Deposited Au/Al2O3 nanocomposite films exhibit superior stability, and average interparticle distances between Au nanoparticles with similar average sizes can be tuned by changing surface coverage. These characteristics are ascribed to the spacer function and rim confinement of dielectric Al2O3 and highlight their advantages for application in optimal nanoparticle‐amplified SPR, especially when the probe size is smaller than the target molecule size. This importance is demonstrated here for the binding of protein (streptavidin) targets to the probe (biotin) surface. In this case, the dielectric matrix Al2O3 is a main contributor, behaving as a spacer, tuning the concentration of Au nanoparticles, and manipulating the average interparticle distance, and thus guaranteeing an appropriate number of biotin molecules and expected near‐field coupling to obtain optimal sensing performance. 相似文献
104.
Masashi Imanaka Shigeo Tomita Suguru Kanda Mitsuteru Fujieda Kimikazu Sasa Jens Olaf Pepke Pedersen Hiroshi Kudo 《Journal of aerosol science》2010,41(5):468-474
To investigate the contribution of ions to gas nucleation, we have performed experiments on the formation of water droplets in H2O/N2 and H2O/Ar gas mixtures by irradiation with a 20 MeV proton beam and by positive corona discharge. The size of the formed nanoparticles was measured using a differential mobility analyzer equipped with a Faraday cup electrometer. Using the proton beam, droplets around 10 nm in diameter were observed for both positively and negatively charged particles, but none were found when the corona discharge was used. This implies the importance of the presence of both positive and negative ions for the formation of nanosize droplets, which attract each other by Coulomb interactions, enhancing the collision frequency and leading to the formation of the 10 nm droplets. 相似文献
105.
Fumitaka Kudo Dr. Atsushi Motegi Kazutoshi Mizoue Dr. Tadashi Eguchi Prof. Dr. 《Chembiochem : a European journal of chemical biology》2010,11(11):1574-1582
FD‐891 is a 16‐membered cytotoxic antibiotic macrolide that is especially active against human leukemia such as HL‐60 and Jurkat cells. We identified the FD‐891 biosynthetic (gfs) gene cluster from the producer Streptomyces graminofaciens A‐8890 by using typical modular type I polyketide synthase (PKS) genes as probes. The gfs gene cluster contained five typical modular type I PKS genes (gfsA, B, C, D, and E), a cytochrome P450 gene (gfsF), a methyltransferase gene (gfsG), and a regulator gene (gfsR). The gene organization of PKSs agreed well with the basic polyketide skeleton of FD‐891 including the oxidation states and α‐alkyl substituent determined by the substrate specificities of the acyltransferase (AT) domains. To clarify the involvement of the gfs genes in the FD‐891 biosynthesis, the P450 gfsF gene was inactivated; this resulted in the loss of FD‐891 production. Instead, the gfsF gene‐disrupted mutant accumulated a novel FD‐891 analogue 25‐O‐methyl‐FD‐892, which lacked the epoxide and the hydroxyl group of FD‐891. Furthermore, the recombinant GfsF enzyme coexpressed with putidaredoxin and putidaredoxin reductase converted 25‐O‐methyl‐FD‐892 into FD‐891. In the course of the GfsF reaction, 10‐deoxy‐FD‐891 was isolated as an enzymatic reaction intermediate, which was also converted into FD‐891 by GfsF. Therefore, it was clearly found that the cytochrome P450 GfsF catalyzes epoxidation and hydroxylation in a stepwise manner in the FD‐891 biosynthesis. These results clearly confirmed that the identified gfs genes are responsible for the biosynthesis of FD‐891 in S. graminofaciens. 相似文献
106.
107.
Akane Samizo Makoto Minohara Naoto Kikuchi Kenta Ando Yuta Mazuka Keishi Nishio 《Journal of the American Ceramic Society》2023,106(2):1540-1546
Sn2Nb2−xTaxO7 (x = 0.0–2.0) with pyrochlore structure is a promising material for p-type oxide semiconductors. A systematic study of its Nb/Ta ratio indicated that the hole–generation efficiency of the Nb end (Sn2Nb2O7) was an order of magnitude lower than that of the Ta end (Sn2Ta2O7). Although this occurs due to differences in oxygen-vacancy formation, the origins of the hole–generation efficiencies remain unclear due to limited information on local and global crystal-structure disorders in pyrochlore Sn2Nb2O7 and Sn2Ta2O7. In this study, the crystal structures of Sn2B2O7 (B = Nb, Ta), composed of BO6 octahedra and Sn4O tetrahedra, were investigated using X-ray absorption spectroscopy and X-ray diffraction. A detailed investigation of the local and global crystal structures indicated a larger amount of disorder in the Sn4O tetrahedra in Sn2Nb2O7 compared to Sn2Ta2O7; disorder in the BO6 octahedra occurred only in Sn2Ta2O7. This study indicates that an appropriate selection of the B-site element is vital for suppressing defect and disorder formation in Sn4O tetrahedra and subsequently improving the hole–carrier–generation efficiency. 相似文献
108.
Ohmori T Morita H Tanaka M Miyauchi K Kasai D Furukawa K Miyashita K Ogawa N Masai E Fukuda M 《Journal of Bioscience and Bioengineering》2011,111(4):437-442
Rhodococcus jostii RHA1 accumulates chlorobenzoates (CBA) during the degradation of polychlorinated biphenyls (PCBs). CBA degradation is considered one of the rate-limiting steps in the complete degradation of PCBs. To reduce the accumulation of CBAs, the upper pathway enzyme genes for PCB degradation of RHA1 were introduced into a CBA-degrading bacterium, Burkholderia sp. NK8. The resulting recombinant strain exhibited no biphenyl 2,3-dioxygenase (BphA) activity encoded by bphAaAbAcAd genes, which encode the large and small subunits of the terminal oxygenase component and the ferredoxin and reductase subunits responsible for electron transfer from NADH to the large subunit. The remaining enzyme genes involved in the transformation of biphenyl to benzoate, bphB2C1D1, which encode dehydrogenase, ring-cleavage dioxygenase and hydrolase, conferred activities to NK8. To obtain the BphA activity of RHA1 in NK8, sets of BphA genes were constructed by combining the bphAaAbAcAd genes of RHA1 and bphA3A4 of Pseudomonas pseudoalcaligenes KF707, encoding the ferredoxin and reductase subunits. Hybrid derivatives of BphA containing the KF707 bphA3 conferred BphA activity to NK8, and a derivative containing the RHA1 bphAaAb and KF707 bphA3A4 genes exhibited the highest BphA activity. A plasmid containing the RHA1 bphAaAb and KF707 bphA3A4 genes plus the RHA1 bphB2C1D1 genes was constructed and introduced into NK8. The resulting recombinant strain efficiently degraded 2-, 3- and 4-chlorobiphenyls with an apparent reduction in CBA accumulation in comparison to the recombinant mutant strain, which had an insertion in the cbeA gene to inactivate CBA dioxygenase. 相似文献
109.
Diah Permata Naoto Nagaoka 《IEEJ Transactions on Electrical and Electronic Engineering》2014,9(Z1):S37-S43
A method to measure a small capacitance with its loss resistance using a set of transient current waveforms is proposed in this paper. The parameters are obtained from the time constants in the time domain. The method has high resistance to extraneous noise, because a time‐to‐frequency transformation, which is sensitive to noise, is not required. The transient current waveforms are obtained by a voltage or current source, a current transformer, and a waveform recorder. The measured capacitance by the proposed method is not affected by the capacitance of the voltage probe, because it is obtained without any voltage information. The sheath surge impedance of the current injection cable, which is indispensable for the transient measurement, is corrected. The application of the method has the advantage in that it allows the modeling of a fast transient of a power apparatus, comparing it with the steady‐state measurement using an impedance measuring instrument. The proposed method is applied to create an equivalent circuit between electrodes implanted into a piece of wood, and its reliability is confirmed by comparison between the measured and calculated results. Stray capacitances of a miniature circuit breaker are also measured, and the results show that the proposed method is applicable to equipment in power systems. © 2014 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc. 相似文献
110.
Naoto Kitamura Takuya Hasegawa Yoshiharu Uchimoto Koji Amezawa Yasushi Idemoto 《Electrochimica acta》2011,(25):9453
LiNi0.5Mn0.5O2 was heat-treated under high oxygen-pressure and Ar-reducing conditions, and then the cathode properties, thermodynamic stability and average and local structures were investigated. From X-ray diffraction and ICP measurements, it was found that the pristine LiNi0.5Mn0.5O2 had a single phase of the layered rock-salt structure although the Ni content was slightly rich compared with the nominal one. These characteristics were kept even after the heat-treatments. Charge–discharge cycle tests clarified that the cycle performance of LiNi0.5Mn0.5O2 was improved by both the reducing and oxidizing treatments. From neutron diffraction and X-ray absorption fine structure measurements, the local distortion around the transition metal, especially Ni, was supposed to be one of the important factors to determine the cathode properties. It was also found that the sample with higher thermodynamic stability exhibited better capacity retention in the discharge–charge cycle tests. 相似文献