Japan Institute of Leather Research

Toride, Japan

Japan Institute of Leather Research

Toride, Japan
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Ogata K.,Japan Institute of Leather Research | Kumazawa Y.,Japan Institute of Leather Research | Koyama Y.,Japan Institute of Leather Research | Yoshimura K.,Tokyo Metropolitan Leather Technology Center | Takahahi K.,Tokyo University of Agriculture and Technology
Journal of the Society of Leather Technologists and Chemists | Year: 2017

Hexavalent chromium (Cr6+) is formed in chrome-tanned leather due to oxidation of trivalent chromium (Cr3+) under certain conditions and may induce dermatitis and cancer. Specifically, significant amounts of Cr6+ are generated during the heat-ageing process by pre-heating at 80°C for 24 hours in accordance with ISO method 17075, promoting the formation of Cr6+ from Cr3+ in chrome-tanned leather. Thus, inhibiting or minimizing the production of Cr6+ is needed badly. A previous report demonstrated that 0.1mol/L potassium phosphate buffer (P-buffer; pH5.5) was highly efficient as an extractant for the measurement of Cr6+ from chrome-tanned leather as compared with P-buffer (pH8.0). The present report describes efforts to control the redox behaviour of Cr3+ with the goal of inhibiting the formation of Cr6+ in chrome-tanned leather using a low-molecular weight (Mw 9,400) collagen peptide (CP-9), 2, 6-di-t-butyl-4-methylphenol (BHT), 3(2)-t-butyl-4-hydroxyanisole (BHA), and ascorbic acid (AsA) as inhibitors, even with heat-aged leather. Chrome-tanned leather was treated with the inhibitors, and the resulting leather samples were extracted with P-buffer (pH5.5) to determine the amount of Cr6+ in the extracts. The results showed that CP-treatment significantly reduced the amount of Cr6+ formed, probably due to a combination of chelation of CP on the Cr3+ complex, quenching of reactive oxygen, and radical scavenging. The BHT, and BHA, and AsA treatments also reduced the amount of Cr6+ formed, probably due to scavenging radicals from the oxidative products of the fatliquoring agent and alterations in the reducing conditions. However, treatment with any of the inhibitors individually did not cause complete inhibition of Cr3+ oxidation. Therefore, the action of the combined inhibitors was investigated. The results demonstrated that treatment with BHA + CP and with BHA + AsA produced a synergistic reduction in Cr6+ production. Use of BHA + AsA + CP completely inhibited the formation of Cr6+ from heat-aged leather samples.


Kumazawa Y.,Japan Institute of Leather Research | Taga Y.,Nippi Research Institute of Biomatrix | Iwai K.,Japan Institute of Leather Research | Koyama Y.-I.,Japan Institute of Leather Research | Koyama Y.-I.,Nippi Research Institute of Biomatrix
Journal of Agricultural and Food Chemistry | Year: 2016

Identification of the animal source of leather is difficult using traditional methods, including microscopic observation and PCR. In the present study, a LC-MS method was developed for detecting interspecies differences in the amino acid sequence of type I collagen, which is a major component of leather, among six animals (cattle, horse, pig, sheep, goat, and deer). After a dechroming procedure and trypsin digestion, six tryptic peptides of type I collagen were monitored by LC-MS in multiple reaction monitoring mode for the animal source identification using the patterns of the presence or absence of the marker peptides. We analyzed commercial leathers from various production areas using this method, and found some leathers in which the commercial label disagreed with the identified animal source. Our method enabled rapid and simple leather certification and could be applied to other animals whether or not their collagen sequences are available in public databases. © 2016 American Chemical Society.


Ogata K.,Japan Institute of Leather Research | Kumazawa Y.,Japan Institute of Leather Research | Koyama Y.,Japan Institute of Leather Research | Yoshimura K.,Tokyo Metropolitan Leather Technology Center | Takahashi K.,Tokyo University of Agriculture and Technology
XXXIII IULTCS Congress | Year: 2015

Hexavalent chromium (Cr6+) induces dermatitis and cancer, and is often quantified from chrome-Tanned leather could be generated by oxidation of trivalent chromium (Cr3+) under a certain condition. In particular, much amount of Cr6+ has been determined by the heat-Aging treatment by pre-heating at 80°C for 24 h according to the method of ISO 17075 due to promoting generation of Cr6+ from Cr3+ in chrome-Tanned leather. It is thus strongly desired that the generation of Cr6+ could be effectively inhibited by a suitable manner. We have previously reported that 0.1 M potassium phosphate buffer (P-buffer; pH 5.5) would be highly efficient as an extractant in measuring Cr6+ from chrome-Tanned leather. In this study, some controlling materials Cr3+ such as stabilizing agent of Cr3+, radical scavenger, and reducing agent, were applied to inhibit generation of Cr6+ for chrome-Tanned leather even if heat-Aged. Collagen peptide (CP-10) with low-molecular weight (Mw about 10,000) 2, 6-di-T-buthyl-4-methylphenol (BHT) and 3(2)-T-buthyl-4-hydroxyanisole (BHA), and ascorbic acid (AsA) were used as a stabilizing agent through reacting with Cr3+, scavenger of radicals generated at the initiation of oxidation of fatliquoring agent, and reducing agent adjusting reducing enviroment, respectively. Chrome-Tanned leather was treated with these controlling materials, and then the resulting leather samples were extracted with P-buffer (pH 5.5) to determine the amount of Cr6+ in the extract. Consequently, CP-Treatment considerably reduced generation of Cr6+ probably due to stabilization of Cr3+ complex with coordination of CP. BHT-and BHA-, and AsA-Treatments also showed considerable reduction probably due to scavenging radical from oxidative products of fatliquoring agent, and due to adjusting reducing environment, respectively. However, the individual treatment with the controlling material could not result in complete inhibition. Combined treatment was thus carried out. Concequently, coupled treatment with BHA + CP and and BHA + AsA exhibited synergy reduction of the Cr6+-generation, in particular, combined treatment with BHA + AsA + CP completely inhibited generation of Cr6+ from the heat-Aged leather sample.


Ueno T.,Nippi Research Institute of Biomatrix | Kaneko K.,Japan National Institute of Infectious Diseases | Katano H.,Japan National Institute of Infectious Diseases | Sato Y.,Japan National Institute of Infectious Diseases | And 10 more authors.
Experimental Cell Research | Year: 2010

A coiled-coil endoplasmic reticulum (ER) protein, p180, was originally reported as a ribosome-binding receptor on the rough ER and is highly expressed in secretory tissues. Recently, we reported new functions of p180 as a microtubule-bundling protein on the ER. Here, we investigated the specific roles of p180 in the Golgi complex organization following stimulated collagen secretion. Targeted depletion of p180 by siRNA transfection caused marked reduction of TGN, while other marker levels for the cis or medial Golgi were not markedly changed. Ascorbate stimulation resulted in trans-Golgi network (TGN) expansion to the periphery in control cells that is characterized by both increased membrane amounts and extended shape. In contrast, loss of p180 resulted in retraction of the TGN regardless of ascorbate stimulation. The TGN developed to the periphery along stabilized microtubule bundles, and overexpression of MTB-1 fragment caused dominant-negative phenotypes. Once disorganized, the retracted TGN did not recover in the absence of p180 despite elevated acetylated tubulin levels. TGN46 and p180 were co-distributed in epithelial basal layer cells of human mucosal and gastrointestinal tissues. Taken together, we propose a novel function of p180-abundant ER on the TGN expansion, both of which are highly developed in various professional secretory cells. © 2009 Elsevier Inc. All rights reserved.


Ueno T.,Nippi Research Institute of Biomatrix | Tanaka K.,Nippi Research Institute of Biomatrix | Kaneko K.,Japan National Institute of Infectious Diseases | Taga Y.,Nippi Research Institute of Biomatrix | And 8 more authors.
Journal of Biological Chemistry | Year: 2010

A coiled-coil microtubule-bundling protein, p180, was originally reported as a ribosome-binding protein on the rough endoplasmic reticulum (ER) and is highly expressed in secretory tissues. Recently, we reported a novel role for p180 in the trans-Golgi network (TGN) expansion following stimulated collagen secretion. Here, we show that p180 plays a key role in procollagen biosynthesis and secretion in diploid fibroblasts. Depletion of p180 caused marked reductions of secreted collagens without significant loss of the ER membrane or mRNA. Metabolic labeling experiments revealed that the procollagen biosynthetic activity was markedly affected following p180 depletion. Moreover, loss of p180 perturbs ascorbate-stimulated de novo biosynthesis mainly in the membrane fraction with a preferential secretion defect of large proteins. At the EM level, one of the most prominent morphological features of p180-depleted cells was insufficient ribosome association on the ER membranes. In contrast, the ER of control cells was studded with numerous ribosomes, which were further enhanced by ascorbate. Similarly biochemical analysis confirmed that levels of membrane-bound ribosomes were altered in a p180-dependent manner. Taken together, our data suggest that p180 plays crucial roles in enhancing collagen biosynthesis at the entry site of the secretory compartments by a novel mechanism that mainly involves facilitating ribosome association on the ER. © 2010 by The American Society for Biochemistry and Molecular Biology, Inc.


Nakayama H.,Wakayama Prefectural Fisheries Experimental Station | Tanaka K.,Nippi Research Institute of Biomatrix | Teramura N.,Nippi Research Institute of Biomatrix | Hattori S.,Nippi Research Institute of Biomatrix | Hattori S.,Japan Institute of Leather Research
Bioscience, Biotechnology and Biochemistry | Year: 2016

The collagenase activity and the fpcol gene were examined in Flavobacterium psychrophilum isolates from cold-water disease (CWD)-affected ayu, Plecoglossus altivelis. Collagenase expression was closely related to the accumulated mortality of CWD-affected ayu. RT-qPCR and bacterial challenge experiments showed that F. psychrophilum ayu isolate WA-1 expressed the fpcol gene more actively and was more virulent than ayu isolate WA-2. The amago (Oncorhynchus masou) isolate WB-1, which possesses a pseudo-fpcol gene, was not harmful to ayu. Hitherto, the well-studied metalloproteases Fpp1 and Fpp2 have been considered virulence factors. However, the most virulent isolate against ayu (WA-1) showed no Fpp activity because of a deletion mutation or an insertion of a transposon in the fpp genes. The less virulent WA-2 isolate showed only Fpp1 activity. Taken together, these results suggest that collagenolytic activity, but not Fpp activity, is related to the virulence of F. psychrophilum isolates in CWD-affected ayu. © 2015 Japan Society for Bioscience, Biotechnology, and Agrochemistry.


Ueno T.,Nippi Research Institute of Biomatrix | Kaneko K.,Japan National Institute of Infectious Diseases | Sata T.,Japan National Institute of Infectious Diseases | Sata T.,Toyama Institute of Health | And 4 more authors.
Nucleic Acids Research | Year: 2012

A coiled-coil microtubule-bundling protein, p180, was originally identified as one of the ribosome receptor candidates on the rough endoplasmic reticulum (ER) and is highly expressed in secretory tissues. Recently, we reported that p180 plays crucial roles in upregulating collagen biosynthesis, mainly by facilitating ribosome association on the ER. Here, we provide evidence that p180 is required to form translationally active polysome/translocon complexes on the ER. Assembly of highly-developed polysomes on the ER was severely perturbed upon loss of p180. p180 associates with polysome/translocon complexes through multiple contact sites: it was coimmunoprecipitated with the translocon complex independently of ribosomes, while it can also bind to ribosomal large subunit specifically. The responsible domain of p180 for membrane polysome assembly was identified in the C-terminal coiled-coil region. The degree of ribosome occupation of collagen and fibronectin mRNAs was regulated in response to increased traffic demands. This effect appears to be exerted in a manner specific for a specified set of mRNAs. Collectively, our data suggest that p180 is required to form translationally active polysome/translocon complexes on the ER membrane, and plays a pivotal role in highly efficient biosynthesis on the ER membrane through facilitating polysome formation in professional secretory cells. © 2011 The Author(s).


Teramura N.,Nippi Research Institute of Biomatrix | Tanaka K.,Nippi Research Institute of Biomatrix | Iijima K.,Nippi Research Institute of Biomatrix | Hayashida O.,Nippi Research Institute of Biomatrix | And 5 more authors.
Journal of Bacteriology | Year: 2011

The collagenase gene was cloned from Grimontia (Vibrio) hollisae 1706B, and its complete nucleotide sequence was determined. Nucleotide sequencing showed that the open reading frame was 2,301 bp in length and encoded an 84-kDa protein of 767 amino acid residues. The deduced amino acid sequence contains a putative signal sequence and a zinc metalloprotease consensus sequence, the HEXXH motif. G. hollisae collagenase showed 60 and 59% amino acid sequence identities to Vibrio parahaemolyticus and Vibrio alginolyticus collagenase, respectively. In contrast, this enzyme showed <20% sequence identity with Clostridium histolyticum collagenase. When the recombinant mature collagenase, which consisted of 680 amino acids with a calculated molecular mass of 74 kDa, was produced by the Brevibacillus expression system, a major gelatinolytic protein band of ~60 kDa was determined by zymographic analysis. This result suggested that cloned collagenase might undergo processing after secretion. Moreover, the purified recombinant enzyme was shown to possess a specific activity of 5,314 U/mg, an ~4-fold greater activity than that of C. histolyticum collagenase. © 2011, American Society for Microbiology. All Rights Reserved.


Patent
Japan Institute Of Leather Research and Osaka University | Date: 2012-08-22

It is an object of the present invention to provide a Type. I-Type IV collagen hybrid gel, which maintains characteristics of a Type IV collagen and is superior in gel strength. It is the Type I-Type IV collagen hybrid gel obtained by mixing 100 to 500 parts by mass of the Type I collagen having fibrosis ability, relative to 100 parts by mass of the Type IV collagen having gelling ability. A three-dimensional structure is formed, where a membrane-like substance by the Type IV collagen is formed onto a fibrous substance by the Type I collagen, so as to be able to provide cell culture environment approximate to a basement membrane of a living body.


PubMed | Japan Institute of Leather Research and Nippi Research Institute of Biomatrix
Type: Journal Article | Journal: Journal of agricultural and food chemistry | Year: 2016

Identification of the animal source of leather is difficult using traditional methods, including microscopic observation and PCR. In the present study, a LC-MS method was developed for detecting interspecies differences in the amino acid sequence of type I collagen, which is a major component of leather, among six animals (cattle, horse, pig, sheep, goat, and deer). After a dechroming procedure and trypsin digestion, six tryptic peptides of type I collagen were monitored by LC-MS in multiple reaction monitoring mode for the animal source identification using the patterns of the presence or absence of the marker peptides. We analyzed commercial leathers from various production areas using this method, and found some leathers in which the commercial label disagreed with the identified animal source. Our method enabled rapid and simple leather certification and could be applied to other animals whether or not their collagen sequences are available in public databases.

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