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161.
Yohei Iizaka Ryusei Arai Akari Takahashi Mikino Ito Miho Sakai Atsushi Fukumoto David H Sherman Yojiro Anzai 《Journal of industrial microbiology & biotechnology》2022,49(1)
MycG is a multifunctional P450 monooxygenase that catalyzes sequential hydroxylation and epoxidation or a single epoxidation in mycinamicin biosynthesis. In the mycinamicin-producing strain Micromonospora griseorubida A11725, very low-level accumulation of mycinamicin V generated by the initial C-14 allylic hydroxylation of MycG is observed due to its subsequent epoxidation to generate mycinamicin II, the terminal metabolite in this pathway. Herein, we investigated whether MycG can be engineered for production of the mycinamicin II intermediate as the predominant metabolite. Thus, mycG was subject to random mutagenesis and screening was conducted in Escherichia coli whole-cell assays. This enabled efficient identification of amino acid residues involved in reaction profile alterations, which included MycG R111Q/V358L, W44R, and V135G/E355K with enhanced monohydroxylation to accumulate mycinamicin V. The MycG V135G/E355K mutant generated 40-fold higher levels of mycinamicin V compared to wild-type M. griseorubida A11725. In addition, the E355K mutation showed improved ability to catalyze sequential hydroxylation and epoxidation with minimal mono-epoxidation product mycinamicin I compared to the wild-type enzyme. These approaches demonstrate the ability to selectively coordinate the catalytic activity of multifunctional P450s and efficiently produce the desired compounds. 相似文献
162.
SMALL ACIDIC PROTEIN 1 (SMAP1) functions upstream of the degradation of AUX/IAA-proteins in the response to 2,4-dichlorophenoxyacetic acid and physically interacts with the COP9 SIGNALOSOME (CSN). Also, its function is linked to RELATED TO UBIQUITIN (RUB) modification. To further investigate the relationship between SMAP1 and the RUB modification system, we examined the effect of MLN4924, an inhibitor of RUB/NEDD8-activating E1 enzyme, on the growth of Arabidopsis thaliana. We found that the anti-auxin resistant 1 mutants, which lack SMAP1, are more sensitive to MLN4924 than wild type and that SMAP1 is responsible for this hypersensitivity. This new evidence supports our previous speculation that SMAP1 acts in Cullin-RING ubiquitin E3 ligase regulated signaling processes via its interaction with components associated with the RUB modification system. 相似文献
163.
Jun Hirose Akari Nagayoshi Naoya Yamanaka Yuji Araki Haruhiko Yokoi 《Biotechnology and Bioprocess Engineering》2013,18(4):736-741
Lignin, a major component of biomass, composed of homogeneous phenolic monomers and functions as a synthetic precursor in the production of specialty chemicals or polymers. In this study, bacterial strains that metabolize lignin-derived low molecular weight compounds (LLCs) were cultured which are capable of LLC bioconversion. We used an LLC mixture primarily composed of vanillin (VL), syringaldehyde (SA), vanillic acid (VA) and p-hydroxybenzoic acid which were prepared from a commercial alkaline lignin product. Enrichment culture was repeated twice in a medium containing the soil sample, the LLCs and inorganic salts. Three bacterial strains belonging to the genera Pseudomonas, Ochrobactrum, and Klebsiella were isolated. We found that only VL, SA, and VA were metabolized by the Pseudomonas strain, which was then found to grow in a medium with VL or VA as the sole source of carbon and energy. The VL isomers, namely, ovanillin and isovanillin were converted to the corresponding carboxylic acids but were not utilized as carbon sources by Pseudomonas. VL and VA are intermediates in the pathway of bacterial degradation of eugenol via ferulic acid. Several bacterial strains that metabolize VL, eugenol, and ferulic acid have been reported but such strains are rarely isolated from enrichment culture medium containing LLCs, due to insufficient induction by the precursors in the LLC medium. In this study, we demonstrated that the microorganisms involved in the bioconversion of LLCs can be isolated from simple enrichment culture. 相似文献
164.
Koutarou Nakamura Seiichiro Sakai Jun Tsuyama Akari Nakamura Kento Otani Kumiko Kurabayashi Yoshiko Yogiashi Hisao Masai Takashi Shichita 《PLoS biology》2021,19(5)
Inflammation is implicated in the onset and progression of various diseases, including cerebral pathologies. Here, we report that DJ-1, which plays a role within cells as an antioxidant protein, functions as a damage-associated molecular pattern (DAMP) and triggers inflammation if released from dead cells into the extracellular space. We first found that recombinant DJ-1 protein induces the production of various inflammatory cytokines in bone marrow–derived macrophages (BMMs) and dendritic cells (BMDCs). We further identified a unique peptide sequence in the αG and αH helices of DJ-1 that activates Toll-like receptor 2 (TLR2) and TLR4. In the ischemic brain, DJ-1 is released into the extracellular space from necrotic neurons within 24 h after stroke onset and makes direct contact with TLR2 and TLR4 in infiltrating myeloid cells. Although DJ-1 deficiency in a murine model of middle cerebral artery occlusion did not attenuate neuronal injury, the inflammatory cytokine expression in infiltrating immune cells was significantly decreased. Next, we found that the administration of an antibody to neutralize extracellular DJ-1 suppressed cerebral post-ischemic inflammation and attenuated ischemic neuronal damage. Our results demonstrate a previously unknown function of DJ-1 as a DAMP and suggest that extracellular DJ-1 could be a therapeutic target to prevent inflammation in tissue injuries and neurodegenerative diseases.Intracellular expression of the antioxidant protein DJ-1 has previously been shown to be neuroprotective. This study reveals that extracellularly released DJ-1 from necrotic neurons is a trigger of sterile inflammation that promotes neuronal injury and neurological deficits after ischemic stroke. 相似文献
165.
166.
Taeko Miyazaki Akari Kato Takanori Ikenaga Hanako Hagio Naoyuki Yamamoto 《Journal of morphology》2019,280(4):526-533
We identified a morphologically uncommon piscine retractor lentis muscle in the yellowfin goby Acanthogobius flavimanus. This lentis muscle has a shape similar to the Greek small letter lambda (λ). The two legs of the muscle are attached to the retinal periphery at the ventral eyecup, while the tip is connected to the lens surface by a ligament. Scanning electron microscopy showed that the fibers of the lentis muscle run along the length of both the anterior and posterior legs. Immunolabeling with antiacetylated tubulin antibody and neuronal tracing with DiI of the whole lentis muscle revealed that the anterior leg is innervated by one or more nerves. The topographic distribution of ganglion cells in the retina was investigated to identify the visual axis. Three high cell density areas were observed in the dorso-temporal, ventro-nasals and ventro-temporal retina. These findings suggest that the λ-shaped lentis muscle may enable accommodatory movement of the lens toward the temporal as well as the nasal and/or ventral retina. 相似文献
167.
Fuyu Kobirumaki-Shimozawa Kotaro Oyama Togo Shimozawa Akari Mizuno Takashi Ohki Takako Terui Susumu Minamisawa Shin’ichi Ishiwata Norio Fukuda 《The Journal of general physiology》2016,147(1):53-62
Sarcomeric contraction in cardiomyocytes serves as the basis for the heart’s pump functions in mammals. Although it plays a critical role in the circulatory system, myocardial sarcomere length (SL) change has not been directly measured in vivo under physiological conditions because of technical difficulties. In this study, we developed a high speed (100–frames per second), high resolution (20-nm) imaging system for myocardial sarcomeres in living mice. Using this system, we conducted three-dimensional analysis of sarcomere dynamics in left ventricular myocytes during the cardiac cycle, simultaneously with electrocardiogram and left ventricular pressure measurements. We found that (a) the working range of SL was on the shorter end of the resting distribution, and (b) the left ventricular–developed pressure was positively correlated with the SL change between diastole and systole. The present findings provide the first direct evidence for the tight coupling of sarcomere dynamics and ventricular pump functions in the physiology of the heart. 相似文献
168.
Yu Yamazaki Chia-Chen Liu Akari Yamazaki Francis Shue Yuka A. Martens Yuanxin Chen Wenhui Qiao Aishe Kurti Hiroshi Oue Yingxue Ren Ying Li Tomonori Aikawa Yesesri Cherukuri John D. Fryer Yan W. Asmann Betty Y.S. Kim Takahisa Kanekiyo Guojun Bu 《Neuron》2021,109(3):438-447.e6
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