Mizkan Group Corporation

Okazaki, Japan

Mizkan Group Corporation

Okazaki, Japan
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Su'etsugu M.,Kyushu University | Su'etsugu M.,Rikkyo University | Harada Y.,Kyushu University | Harada Y.,Mizkan Group Corporation | And 7 more authors.
Environmental Microbiology | Year: 2013

DnaA activity for replication initiation of the Escherichia coli chromosome is negatively regulated by feedback from the DNA-loaded form of the replicase clamp. In this process, called RIDA (regulatory inactivation of DnaA), ATP-bound DnaA transiently assembles into a complex consisting of Hda and the DNA-clamp, which promotes inter-AAA+ domain association between Hda and DnaA and stimulates hydrolysis of DnaA-bound ATP, producing inactive ADP-DnaA. Using a truncated DnaA mutant, we previously demonstrated that the DnaA N-terminal domain is involved in RIDA. However, the precise role of the N-terminal domain in RIDA has remained largely unclear. Here, we used an in vitro reconstituted system to demonstrate that the Asn-44 residue in the N-terminal domain of DnaA is crucial for RIDA but not for replication initiation. Moreover, an assay termed PDAX (pull-down after cross-linking) revealed an unstable interaction between a DnaA-N44A mutant and Hda. In vivo, this mutant exhibited an increase in the cellular level of ATP-bound DnaA. These results establish a model in which interaction between DnaA Asn-44 and Hda stabilizes the association between the AAA+ domains of DnaA and Hda to facilitate DnaA-ATP hydrolysis during RIDA. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

Fukuda M.,Tohoku University | Watanabe S.,Tohoku University | Watanabe S.,Mizkan Group Co. | Yoshida S.,University of Tsukuba | And 4 more authors.
Journal of Bacteriology | Year: 2010

Paenibacillus sp. W-61 is capable of utilizing water-insoluble xylan for carbon and energy sources and has three xylanase genes, xyn1, xyn3, and xyn5. Xyn1, Xyn3, and Xyn5 are extracellular enzymes of the glycoside hydrolase (GH) families 11, 30, and 10, respectively. Xyn5 contains several domains including those of carbohydrate-binding modules (CBMs) similar to a surface-layer homologous (SLH) protein. This study focused on the role of Xyn5, localized on the cell surface, in water-insoluble xylan utilization. Electron microscopy using immunogold staining revealed Xyn5 clusters over the entire cell surface. Xyn5 was bound to cell wall fractions through its SLH domain. A Δxyn5 mutant grew poorly and produced minimal amounts of Xyn1 and Xyn3 on water-insoluble xylan. A Xyn5 mutant lacking the SLH domain (Xyn5ΔSLH) grew poorly, secreting Xyn5ΔSLH into the medium and producing minimal Xyn1 and Xyn3 on water-insoluble xylan. A mutant with an intact xyn5 produced Xyn5 on the cell surface, grew normally, and actively synthesized Xyn1 and Xyn3 on water-insoluble xylan. Quantitative reverse transcription-PCR showed that xylobiose, generated from water-insoluble xylan decomposition by Xyn5, is the most active inducer for xyn1 and xyn3. Luciferase assays using a Xyn5-luciferase fusion protein suggested that xylotriose is the best inducer for xyn5. The cell surface Xyn5 appears to play two essential roles in water-insoluble xylan utilization: (i) generation of the xylo-oligosaccharide inducers of all the xyn genes from water-insoluble xylan and (ii) attachment of the cells to the substrate so that the generated inducers can be immediately taken up by cells to activate expression of the xyn system. Copyright © 2010, American Society for Microbiology. All Rights Reserved.

Fukami H.,Mizkan Group Corporation | Tachimoto H.,Mizkan Group Corporation | Kishi M.,Mizkan Group Corporation | Kaga T.,Mizkan Group Corporation | And 3 more authors.
Journal of Lipid Research | Year: 2010

We prepared 2-hydroxypalmitoyl-sphinganine (dihydroceramide) labeled with a stable isotope by culturing acetic acid bacteria with 13C-labeled acetic acid. The GC/MS spectrum of the trimethylsilyl derivative of 13C-labeled dihydroceramide gave molecular ions with an increased mass of 12-17 Da over that of nonlabeled dihydroceramide. The fragment ions derived from both sphinganine base and 2-hydroxypalmitate were confirmed to be labeled with the stable isotope in the spectrum. Therefore, 13C- labeled dihydroceramide can be an extremely useful tool for analyzing sphingolipid metabolism. The purified [13C]dihydroceramide was administered orally to mice for 12 days, and the total sphingoid base fractions in various tissues were analyzed by GC/MS. The spectrum patterns specific to 13C-labeled sphingoids were detected in the tissues tested. Sphinganine pools in skin epidermis, liver, skeletal muscle, and synapse membrane in brain were replaced by [13C]sphinganine at about 4.5, 4.0, 1.0, and 0.3%, respectively. Moreover, about 1.0% of the sphingosine pool in the liver was replaced by [13C]sphingosine, implying that exogenous dihydroceramide can be converted to sphingosine. These results clearly indicate that ingested dihydroceramide can be incorporated into various tissues, including brain, and metabolized to other sphingolipids. Copyright © 2010 by the American Society for Biochemistry and Molecular Biology, Inc.

Fukami H.,Mizkan Group Corporation | Tachimoto H.,Mizkan Group Corporation | Kishi M.,Mizkan Group Corporation | Kaga T.,Mizkan Group Corporation | Tanaka Y.,Tokyo Metropolitan University
Journal of Agricultural and Food Chemistry | Year: 2010

Acetic acid bacteria, fermentative microorganisms of traditional foods, have unique alkali-stable lipids (ASL), such as dihydroceramide which is a precursor of sphingolipids. Sphingolipids are important components of the brain tissue. We examined the effect of oral administration of ASL in a rat model of dementia (7-week-old, male) with a basal forebrain lesion. In a water maze test, the dementia model rats demonstrated poor spatial orientation. The administration of ASL (165 or 1650 mg/kg of body weight per day, for 14 days) produced a significant improvement in learning ability in the dementia model rats. In vitro experiments showed ASL had the ability to promote neurite outgrowth in pheochromocytoma (PC12) cells. Among the ASL components, dihydroceramide has the most potent effect on the differentiation of PC12 cells. It is highly possible that oral administration of dihydroceramide-containing ASL reverses the decline in cognitive function in dementia. © 2010 American Chemical Society.

A natto (fermented soybeans) container 11 is made of a resin-foam sheet of a structure of which a lid body, having a natto-holding recess, is connected by a hinge part to, and covering the opening of the container body. A seasoning-holding recess is provided adjacent to the inner-lid surface of the lid body. A pre-defined folding line L1 is described on the outer-lid surface of the lid body, along which the lid body folds upward into a valley shape. A splittable groove, provided on the lid body, communicates with the natto seasoning-holding recesses and crosses the pre-defined folding line L1. A sealant material is provided on the outer-lid surface, covering and sealing the seasoning holding recess. Hence, the lid body folds into a valley-shape along the pre-defined folding-line crossing, thus breaking the splittable groove.

Onoue N.,Hokkaido University | Yamashita Y.,Hokkaido University | Nagao N.,Hokkaido University | Nagao N.,Mizkan Group Corporation | And 4 more authors.
Journal of Biological Chemistry | Year: 2011

Expression of the Arabidopsis CGS1 gene, encoding the first committed enzyme of methionine biosynthesis, is feedback-regulated in response to S-adenosyl-L-methionine (AdoMet) at the mRNA level. This regulation is first preceded by temporal arrest of CGS1 translation elongation at the Ser-94 codon. AdoMet is specifically required for this translation arrest, although the mechanism by which AdoMet acts with the CGS1 nascent peptide remained elusive. We report here that the nascent peptide of CGS1 is induced to form a compact conformation within the exit tunnel of the arrested ribosome in an AdoMet-dependent manner. Cysteine residues introduced into CGS1 nascent peptide showed reduced ability to react with polyethyleneglycol maleimide in the presence of AdoMet, consistent with a shift into the ribosomal exit tunnel. Methylation protection and UV cross-link assays of 28 S rRNA revealed that induced compaction of nascent peptide is associated with specific changes in methylation protection and UV cross-link patterns in the exit tunnel wall. A 14-residue stretch of amino acid sequence, termed the MTO1 region, has been shown to act in cis for CGS1 translation arrest and mRNA degradation. This regulation is lost in the presence of mto1 mutations, which cause single amino acid alterations within MTO1. In this study, both the induced peptide compaction and exit tunnel change were found to be disrupted by mto1 mutations. These results suggest that the MTO1 region participates in the AdoMet-induced arrest of CGS1 translation by mediating changes of the nascent peptide and the exit tunnel wall. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

Arai T.,Kyushu University | Arai T.,Mizkan Group Corporation | Ohkuri T.,Kyushu University | Yasumatsu K.,Kyushu University | And 2 more authors.
Neuroscience | Year: 2010

The transient receptor potential vanilloid-1 (TRPV1) receptor acts as a polymodal nociceptor activated by capsaicin, heat, and acid. TRPV1, which is expressed in sensory neurons innervating the oral cavity, is associated with an oral burning sensation in response to spicy food containing capsaicin. However, little is known about the involvement of TRPV1 in responses to acid stimuli in either the gustatory system or the general somatosensory innervation of the oropharynx. To test this possibility, we recorded electrophysiological responses to several acids (acetic acid, citric acid and HCl) and other taste stimuli from the mouse chorda tympani, glossopharyngeal and superior laryngeal nerves, and compared potential effects of iodo-resiniferatoxin (I-RTX), a potent TRPV1 antagonist, on chemical responses of the three nerves. The results indicated that in the chorda tympani nerve, I-RTX (1-100 nM) did not affect responses to acids, sucrose and quinine HCl, but reduced responses to NaCl (I-RTX at concentrations of 10 and 100 nM) and KCl and NH4Cl (100 nM). In contrast, in the glossopharyngeal nerve, I-RTX significantly suppressed responses to all acids and salts, but not to sucrose and quinine HCl. Responses to acetic acid were suppressed by I-RTX even at 0.1 nM concentration. The superior laryngeal nerve responded in a concentration-dependent manner to acetic acid, citric acid, HCl, KCl, NH4Cl and monosodium l-glutamate. The responses to acetic acid, but not to the other stimuli, were significantly inhibited by I-RTX. These results suggested that TRPV1 may be involved in the mechanism for responses to acids presented to the posterior oral cavity and larynx. This high degree of responsiveness to acetic acid may account for the oral burning sensation, known as a flavor characteristic of vinegar. © 2010 IBRO.

Ishii S.,Mizkan Group Co. | Ishii S.,Tokyo University of Science | Kurokawa A.,Tokyo University of Science | Kishi M.,Mizkan Group Co. | And 4 more authors.
FEBS Journal | Year: 2012

Polycystic kidney disease (PKD) 2L1 protein is a member of the transient receptor potential (TRP) ion channel family. In circumvallate and foliate papillae, PKD2L1 is coexpressed with PKD1L3. PKD2L1 and PKD1L3 interact through their transmembrane domain and the resulting heteromer PKD1L3/PKD2L1 owns a unique channel property called 'off-responses' to acid stimulation, although PKD2L1 does not own this property by itself. To define the pharmacological properties of the PKD1L3/PKD2L1 channel, we developed a new method to effectively evaluate channel activity using human embryonic kidney 293T cells in which the channel was heterologously expressed. This method was applied to screen substances that potentially regulate it. We found that capsaicin and its analogs, which are TRPV1 agonists, inhibited the response to acid stimuli and that the capsaicin inhibition was reversible with an IC 50 of 32.5 μm. Capsaicin and its analogs are thus useful tools for physiological analysis of PKD1L3/PKD2L1 function. Database -Nucleotide sequence data are available in the GenBank database under the accession numbers hTRPA1, To define the pharmacological properties of polycystic kidney disease (PKD) 1L3/PKD2L1 channel, we developed a new method to effectively evaluate channel activity using human embryonic kidney 293T cells. This method was applied and we found that capsaicin and its analogs inhibited the response to acid stimuli and that the capsaicin inhibition was reversible with an IC 50 of 32.5 μm. © 2012 The Authors Journal compilation © 2012 FEBS.

Kada S.,Mizkan Group Co. | Ishikawa A.,Mizkan Group Co. | Ohshima Y.,Mizkan Group Co. | Yoshida K.-I.,Kobe University
Bioscience, Biotechnology and Biochemistry | Year: 2013

Natto is a traditional Japanese food made from soybeans fermented by natto starter strains of Bacillus subtilis natto. It has been suggested that extracellular protease activity released by the bacteria are involved in the production of poly-γ-glutamate (γ-PGA) during natto fermentation. One of the natto starters, strain r22, possesses at least seven genes, each of which encoded an extracellular protease orthologous to its counterpart in B. subtilis 168, aprE, bpr, epr, mpr, nprE, vpr, and wprA, but it was found to lack nprB. Inactivating the aprE ortholog alone resulted in a severe decrease in γ-PGA production and in the total extracellular protease activity. The defect in γ-PGA production of the mutant lacking the aprE ortholog was complemented when the medium was supplemented with sufficient glutamate. These results suggest that the alkaline serine protease encoded by aprE plays an indispensable role in supplying materials to produce γ-PGA. On the other hand, simultaneous inactivation of all the protease genes except for aprE did not significantly affect either γ-PGA production or total protease activity.

Ogawa S.,Mizkan Group Co. | Tachimoto H.,Mizkan Group Co. | Kaga T.,Mizkan Group Co.
Journal of Bioscience and Bioengineering | Year: 2010

Acetic acid bacteria have unique and highly pure membrane lipid components, such as 2-hydroxypalmitoyl-sphinganine (dihydroceramide) and can grow and produce acetic acid at around pH 3.0, suggesting that ceramide in cell membranes may be involved in the tolerance to acidic pH. Acetobacter malorum S24 was selected for the production of ceramide and grown in YPG medium containing 0.8% ethanol. Ceramide biosynthesis was induced at pH 4 and below, suggesting that ceramide biosynthesis is induced by low pH stress. Elevation of ceramide was also induced by high temperature stress (40-70 °C). After the strain was cultured in an optimal growth medium, the cells were collected and treated at pH 3 and 40 °C for 4 days, resulting in a 30-fold elevation of both the yield and content of ceramide. © 2009.

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