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31.
Won-Kyung Hong Sun-Yeon Heo Baek-Rock Oh Chul Ho Kim Jung-Hoon Sohn Ji-Won Yang Akihiko Kondo Jeong-Woo Seo 《Bioprocess and biosystems engineering》2013,36(9):1191-1197
In the present study, we established a genetic system for manipulating the oleaginous heterotrophic microalgae Aurantiochytrium sp. KRS101, using cycloheximide resistance as the selectable marker. The gene encoding ribosomal protein L44 (RPL44) of Aurantiochytrium sp. KRS101 was first identified and characterized. Proline 56 was replaced with glutamine, affording cycloheximide resistance to strains encoding the mutant protein. This resistance served as a novel selection marker. The gene encoding the Δ12-fatty acid desaturase of Mortierella alpina, used as a reporter, was successfully introduced into chromosomal DNA of Aurantiochytrium sp. KRS101 via 18S rDNA-targeted homologous recombination. Enzymatic conversion of oleic acid (C18:1) to linoleic acid (C18:2) was detected in transformants but not in the wild-type strain. 相似文献
32.
Akihiko Shimizu Hiroyuki Tsukagoshi Tsuyoshi Sekizuka Makoto Kuroda Aya Koizumi Masahiro Fujita Yoshiyuki Yamada Nobuhiro Saruki 《Microbiology and immunology》2020,64(9):630-634
Group B streptococcus (GBS) is a leading cause of neonatal infections. Most isolates are β-hemolytic, and their activity is considered to be pivotal for GBS pathogenicity. We report a case of a neonate with meningitis caused by nonhemolytic GBS. The patient developed meningitis 3 days after birth. Genotyping was performed and the characteristics of the strain (GCMC97051) identified by whole genome sequence using next generation sequencing. GCMC97051 possesses genetic alterations such as disruption of cylA by IS1381A insertion and a frameshift mutation in cylE, resulting in a lack of hemolysis. Thus, nonhemolytic GBS can retain the potential to cause invasive infections. 相似文献
33.
Hiroko Iijima Atsuko Watanabe Haruna Sukigara Tomokazu Shirai Akihiko Kondo Takashi Osanai 《Biotechnology and bioengineering》2020,117(6):1649-1660
Synechocystis sp. PCC 6803, a cyanobacterium widely used for basic research, is often cultivated in a synthetic medium, BG-11, in the presence of 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid (HEPES) or 2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid buffer. Owing to the high cost of HEPES buffer (96.9% of the total cost of BG-11 medium), the biotechnological application of BG-11 is limited. In this study, we cultured Synechocystis sp. PCC 6803 cells in BG-11 medium without HEPES buffer and examined the effects on the primary metabolism. Synechocystis sp. PCC 6803 cells could grow in BG-11 medium without HEPES buffer after adjusting for nitrogen sources and light intensity; the production rate reached 0.54 g cell dry weight·L−1·day−1, exceeding that of commercial cyanobacteria and Synechocystis sp. PCC 6803 cells cultivated under other conditions. The exclusion of HEPES buffer markedly altered the metabolites in the central carbon metabolism; particularly, the levels of compatible solutes, such as sucrose, glucosylglycerol, and glutamate were increased. Although the accumulation of sucrose and glucosylglycerol under high salt conditions is antagonistic to each other, these metabolites accumulated simultaneously in cells grown in the cost-effective medium. Because these metabolites are used in industrial feedstocks, our results reveal the importance of medium composition for the production of metabolites using cyanobacteria. 相似文献
34.
Chie Tomikawa Takayuki Ohira Yasushi Inoue Takuya Kawamura Akihiko Yamagishi Tsutomu Suzuki Hiroyuki Hori 《FEBS letters》2013
Thermoplasma acidophilum is a thermo-acidophilic archaeon. We purified tRNALeu (UAG) from T. acidophilum using a solid-phase DNA probe method and determined the RNA sequence after determining via nucleoside analysis and m7G-specific aniline cleavage because it has been reported that T. acidophilum tRNA contains m7G, which is generally not found in archaeal tRNAs. RNA sequencing and liquid chromatography–mass spectrometry revealed that the m7G modification exists at a novel position 49. Furthermore, we found several distinct modifications, which have not previously been found in archaeal tRNA, such as 4-thiouridine9, archaeosine13 and 5-carbamoylmethyuridine34. The related tRNA modification enzymes and their genes are discussed. 相似文献
35.
Koji Ochiai Satoshi Takita Akihiko Kojima Tomohiko Eiraku Kazuhiko Iwase Tetsuya Kishi Akira Ohinata Yuichi Yageta Tokutaro Yasue David R. Adams Yasushi Kohno 《Bioorganic & medicinal chemistry letters》2013,23(1):375-381
(?)-6-(7-Methoxy-2-(trifluoromethyl)pyrazolo[1,5-a]pyridin-4-yl)-5-methyl-4,5-dihydropyridazin-3(2H)-one (KCA-1490) exhibits moderate dual PDE3/4-inhibitory activity and promises as a combined bronchodilatory/anti-inflammatory agent. N-alkylation of the pyridazinone ring markedly enhances potency against PDE4 but suppresses PDE3 inhibition. Addition of a 6-aryl-4,5-dihydropyridazin-3(2H)-one extension to the N-alkyl group facilitates both enhancement of PDE4-inhibitory activity and restoration of potent PDE3 inhibition. Both dihydropyridazinone rings, in the core and extension, can be replaced by achiral 4,4-dimethylpyrazolone subunits and the core pyrazolopyridine by isosteric bicyclic heteroaromatics. In combination, these modifications afford potent dual PDE3/4 inhibitors that suppress histamine-induced bronchoconstriction in vivo and exhibit promising anti-inflammatory activity via intratracheal administration. 相似文献
36.
Akihiko Kojima Satoshi Takita Tatsunobu Sumiya Koji Ochiai Kazuhiko Iwase Tetsuya Kishi Akira Ohinata Yuichi Yageta Tokutaro Yasue Yasushi Kohno 《Bioorganic & medicinal chemistry letters》2013,23(19):5311-5316
We previously identified KCA-1490 [(?)-6-(7-methoxy-2-trifluoromethyl-pyrazolo[1,5-a]pyridin-4-yl)-5-methyl-4,5-dihydro-3-(2H)-pyridazinone], a dual PDE3/4 inhibitor. In the present study, we found highly potent selective PDE4 inhibitors derived from the structure of KCA-1490. Among them, N-(3,5-dichloropyridin-4-yl)-7-methoxy-2-(trifluoromethyl)pyrazolo[1,5-a]pyridine-4-carboxamide (2a) had good anti-inflammatory effects in an animal model. 相似文献
37.
38.
Tateki Hayashi Akihiko Terao Shinji Ueda Mitsuo Namiki 《Bioscience, biotechnology, and biochemistry》2013,77(11):3139-3144
Ascorbic acid-protein mixtures of low moisture content stored for 2~3 days in aerobic conditions at 60°C produced a red coloration, which was shown to have resulted from an amino-carbonyl reaction of oxidized ascorbic acid (dehydroascorbic acid, DHA) by the facts that DHA-casein or ovalbumin systems yielded a rapid red coloration in low moisture conditions as well as in an ethanol suspension. Zein showed only a weak red coloration with DHA. An apparent decrease in the free amino group, as determined by the TNBS method, was observed to be associated in parallel with the red pigment formation. The electrophoretic pattern of ovalbumin changed significantly upon incubation with DHA, together with the formation of the red pigment. The red pigment extracted from a DHA-casein system showed an absorption spectrum, TLC Rf value, and hydrolyzed products (DHA and scorbamic acid) identical to those of 2,2′-nitrilo di-2(2)-deoxy-l-ascorbic acid mono ammonium salt, produced from DHA and amino acid. Formation of the red pigment was also observed in the reaction of DHA with Nα-acetyl lysine. These results indicate that the ε-amino group of lysine in protein can be attributed to the formation of a red pigment identical with NDA. 相似文献
39.
Ryosuke Fujiwara Shuhei Noda Yoshifumi Kawai Tsutomu Tanaka Akihiko Kondo 《Biotechnology letters》2016,38(9):1543-1549
Objectives
To find a novel host for the production of 4-vinylphenol (4VPh) by screening Streptomyces species.Results
The conversion of p-coumaric acid (pHCA) to 4VPh in Streptomyces mobaraense was evaluated using a medium containing pHCA. S. mobaraense readily assimilated pHCA after 24 h of cultivation to produce 4VPh. A phenolic acid decarboxylase, derived from S. mobaraense (SmPAD), was purified following heterologous expression in Escherichia coli. SmPAD was evaluated under various conditions, and the enzyme’s kcat/Km value was 0.54 mM ?1 s?1. Using intergenetic conjugation, a gene from Rhodobacter sphaeroides encoding a tyrosine ammonia lyase, which catalyzes the conversion of l-tyrosine to p-coumaric acid, was introduced into S. mobaraense. The resulting S. mobaraense transformant produced 273 mg 4VPh l?1 from 10 g glucose l?1.Conclusion
A novel strain suitable for the production of 4VPh and potentially other aromatic compounds was isolated.40.
Kengo Sasaki Jun Inoue Daisuke Sasaki Namiko Hoshi Tomokazu Shirai Itsuko Fukuda Takeshi Azuma Akihiko Kondo Ro Osawa 《Biotechnology journal》2019,14(5)
Compositional alteration of the gut microbiota is associated with ulcerative colitis (UC). Here, a model culture system is established for the in vitro human colonic microbiota of UC, which will be helpful for determining medical interventions. 16S ribosomal RNA sequencing confirms that UC models are successfully developed from fecal inoculum and retain the bacterial species biodiversity of UC feces. The UC models closely reproduce the microbial components and successfully preserve distinct clusters from the healthy subjects (HS), as observed in the feces. The relative abundance of bacteria belonging to the family Lachnospiraceae significantly decreases in the UC models compared to that in HS, as observed in the feces. The system detects significantly lower butyrogenesis in the UC models than that in HS, correlating with the decreased abundance of Lachnospiraceae. Interestingly, the relative abundance of Lachnospiraceae does not correlate with disease activity (defined as partial Mayo score), suggesting that Lachnospiraceae persists in UC patients at a decreased level, irrespective of the alteration in disease activity. Moreover, the system shows that administration of Clostridium butyricum MIYAIRI restores butyrogenesis in the UC model. Hence, the model detects deregulation in the intestinal environment in UC patients and may be useful for simulating the effect of probiotics. 相似文献