Biomaterial in Tokyo Co.

Kashiwanoha, Japan

Biomaterial in Tokyo Co.

Kashiwanoha, Japan
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Sato T.,Biomaterial in Tokyo Co. | Sato T.,Kyoto Prefectural University | Yasuzawa T.,Kyoto Prefectural University | Uesaka A.,Kyoto Prefectural University | And 6 more authors.
Nutrition Research | Year: 2014

A precursor of protoporphyrin IX, 5-aminolevulinic acid (5-ALA) is used as a prodrug for photodiagnosis and photodynamic therapy. Recently, it has been shown that 5-ALA reduces glucose levels during fasting and after glucose loading in prediabetic subjects. We hypothesized that 5-ALA ameliorates diabetic conditions through mitochondrial changes in visceral adipose tissue. In order to explore the metabolic effects on the type 2 diabetic state, we administered ALA hydrochloride in combination with sodium ferrous citrate to Otsuka Long-Evans Tokushima Fatty (OLETF) rats at intragastric doses of 20 and 300 mg kg-1 d-1 for 6 weeks. The administration of 300 mg kg-1 d-1 of 5-ALA improved glucose intolerance, hypertriglyceridemia, and hyperleptinemia in OLETF rats more effectively than the administration of an equivalent dose of metformin, in accordance with reductions in food intake and body weight. Furthermore, the weight of the retroperitoneal fat tended to decrease and cellular mitochondrial content of the fat was markedly reduced by the 5-ALA administration, showing a positive correlation. These results suggest that 5-ALA ameliorates diabetic abnormalities in OLETF rats by reducing the visceral fat mass and mitochondrial content of adipocytes in a site-specific manner. © 2014 Elsevier Inc.


PubMed | Kyoto Prefectural University, SBI Pharmaceuticals Co and Biomaterial in Tokyo Co
Type: Journal Article | Journal: Nutrition research (New York, N.Y.) | Year: 2014

A precursor of protoporphyrin IX, 5-aminolevulinic acid (5-ALA) is used as a prodrug for photodiagnosis and photodynamic therapy. Recently, it has been shown that 5-ALA reduces glucose levels during fasting and after glucose loading in prediabetic subjects. We hypothesized that 5-ALA ameliorates diabetic conditions through mitochondrial changes in visceral adipose tissue. In order to explore the metabolic effects on the type 2 diabetic state, we administered ALA hydrochloride in combination with sodium ferrous citrate to Otsuka Long-Evans Tokushima Fatty (OLETF) rats at intragastric doses of 20 and 300 mg kg(-1) d(-1) for 6 weeks. The administration of 300 mg kg(-1) d(-1) of 5-ALA improved glucose intolerance, hypertriglyceridemia, and hyperleptinemia in OLETF rats more effectively than the administration of an equivalent dose of metformin, in accordance with reductions in food intake and body weight. Furthermore, the weight of the retroperitoneal fat tended to decrease and cellular mitochondrial content of the fat was markedly reduced by the 5-ALA administration, showing a positive correlation. These results suggest that 5-ALA ameliorates diabetic abnormalities in OLETF rats by reducing the visceral fat mass and mitochondrial content of adipocytes in a site-specific manner.


Koganei M.,Meiji Co. | Saitou Y.,Meiji Co. | Tsuchiya K.,SBI Pharma Co. | Abe F.,SBI Pharma Co. | And 5 more authors.
Journal of Clinical Biochemistry and Nutrition | Year: 2015

The effects of 5-amino levulinic acid (5-ALA) on obesity were investigated using a murine model (diet-induced obese mice). Diet-induced obese mice were divided into 4 groups: a control group (C group), which was fed a high-fat diet; a low-5-ALA dose (10 mg/kg/day) group (10A group); a moderate-5-ALA dose (30 mg/kg/day) group (30A group); and a high-5-ALA dose (100 mg/kg/day) group (100A group). 5-ALA was administered by mixing the high fat diet for 8 weeks. Body weight increases in the 30A and 100A groups were significantly smaller compared with those of the C group. Body fat measurements by X-ray computed tomography indicated that the 100A group showed a tendency toward low visceral fat quantities during the final week of the study. Visceral fat weights in the 30A and 100A groups were slightly low. The levels of serum alanine aminotransferase (ALT) and total cholesterol (TC) in the 10A group was slightly low, whereas the 30A and 100A groups showed significantly lower ALT and TC values. Liver lipid concentration showed a dose-dependent decrease with ALA. Thus, in this diet-induced obese murine model, administration of 5-ALA had a significantly beneficial impact on the visceral fat, serum ALT and TC, and liver lipid concentration. Copyright © 2015 JCBN.


PubMed | Biomaterial in Tokyo Co., Meiji Co. and SBI Pharma Co.
Type: Journal Article | Journal: Journal of clinical biochemistry and nutrition | Year: 2015

The effects of 5-aminolevulinic acid (5-ALA) on obesity were investigated using a murine model (diet-induced obese mice). Diet-induced obese mice were divided into 4 groups: a control group (C group), which was fed a high-fat diet; a low-5-ALA dose (10mg/kg/day) group (10A group); a moderate-5-ALA dose (30mg/kg/day) group (30A group); and a high-5-ALA dose (100mg/kg/day) group (100A group). 5-ALA was administered by mixing the high fat diet for 8 weeks. Body weight increases in the 30A and 100A groups were significantly smaller compared with those of the C group. Body fat measurements by X-ray computed tomography indicated that the 100A group showed a tendency toward low visceral fat quantities during the final week of the study. Visceral fat weights in the 30A and 100A groups were slightly low. The levels of serum alanine aminotransferase (ALT) and total cholesterol (TC) in the 10A group was slightly low, whereas the 30A and 100A groups showed significantly lower ALT and TC values. Liver lipid concentration showed a dose-dependent decrease with ALA. Thus, in this diet-induced obese murine model, administration of 5-ALA had a significantly beneficial impact on the visceral fat, serum ALT and TC, and liver lipid concentration.


Tachioka M.,University of Tokyo | Sugimoto N.,University of Tokyo | Sugimoto N.,Biomaterial in Tokyo Co. | Nakamura A.,University of Tokyo | And 6 more authors.
Biotechnology for Biofuels | Year: 2016

Background: Random mutagenesis is a powerful technique to obtain mutant proteins with different properties from the wild-type molecule. Error-prone PCR is often employed for random mutagenesis in bacterial protein expression systems, but has rarely been used in the methylotrophic yeast Pichia pastoris system, despite its significant advantages, mainly because large (μg-level) amounts of plasmids are required for transformation. Results: We developed a quick and easy technique for random mutagenesis in P. pastoris by sequential Phi29 DNA polymerase-based amplification methods, error-prone rolling circle amplification (RCA) and multiple displacement amplification (MDA). The methodology was validated by applying it for random mutation of the gene encoding cellulase from the basidiomycete Phanerochaete chrysosporium (PcCel6A), a key enzyme in degradation of cellulosic biomass. In the error-prone RCA step, the concentrations of manganese ion (Mn2+) and cellulase gene-containing plasmid were varied, and the products obtained under each condition were subjected to the second MDA step in the absence of Mn2+. The maximum error rate was 2.6 mutations/kb, as evaluated from the results of large-scale sequencing. Several μg of MDA products was transformed by electroporation into Pichia cells, and the activities of extracellularly expressed PcCel6A mutants towards crystalline and amorphous celluloses were compared with those of wild-type enzyme to identify key amino acid residues affecting degradation of crystalline cellulose. Conclusions: We present a rapid and convenient random mutagenesis method that does not require laborious steps such as ligation, cloning, and synthesis of specific primers. This method was successfully applied to the protein expression system in P. pastoris. © 2016 The Author(s).


Ohta K.,University of Miyazaki | Hamasuna H.,University of Miyazaki | Tsukamoto J.,University of Miyazaki | Wakiyama M.,University of Miyazaki | And 2 more authors.
Journal of Bioscience and Bioengineering | Year: 2012

Ethanol-producing . Escherichia coli strain KO11 consumed 99% of the glucose and only 13% of the xylose in a mixture of glucose (60. g/L) and xylose (40. g/L) during the 72-h fermentation at 30°C. The deletion mutants δ. ptsG, δ. manXYZ, and δ. ptsG/manXYZ utilized 42%, 78%, and 35% of the glucose and 50%, 32%, and 32% of the xylose, respectively. © 2012 The Society for Biotechnology, Japan.


Saga K.,Biomaterial in Tokyo Co. | Hasegawa F.,University of Tokyo | Miyagi S.,University of Tokyo | Atobe S.,University of Tokyo | And 4 more authors.
Applied Energy | Year: 2015

This study aimed to document the material balance and evaluate the energy profit ratios (EPRs) of two processes for recovering hydrocarbon from the green microalgae Botryococcus braunii, which produces hydrocarbon intercellularly, releases it extracellularly, and accumulates it in the extracellular matrix in colony-form. The first is a wet process that elutes an extraction inhibitor through heat, eliminates it through solid-liquid separation, and extracts a hydrocarbon using hexane. The second is a dry process that extracts hydrocarbon by hexane after drying. The EPR of the wet process (3.5) was approximately twice that of the dry process (1.8). Therefore, hydrocarbon can be efficiently recovered from B. braunii without drying. © 2014 Elsevier Ltd.


Atobe S.,University of Tokyo | Saga K.,Biomaterial in Tokyo Co. | Hasegawa F.,University of Tokyo | Furuhashi K.,University of Tokyo | And 4 more authors.
Algal Research | Year: 2015

Water-soluble polymers are released from the extracellular matrix of Botryococcus braunii by thermal pretreatment prior to hydrocarbon extraction. In this study, hydrocarbon recovery was reduced by adding polymers to algal slurries from which they were previously washed. The water-soluble polymers are amphiphilic and emulsify the water-organic solvent systems used in hydrocarbon extraction. To obtain >90% hydrocarbon yields from high-concentration slurry, the original 10% water-soluble polymer content of dry algal cells had to be reduced to less than 0.5%. The water-soluble polymers were polysaccharides with a molecular weight greater than 2×106 and mainly comprised of galactose, arabinose, and uronic acid. We suggest that high-molecular-weight water-soluble polymers are desirable as industrial emulsifiers and thickeners. © 2015.


Chisada S.-I.,Kyushu University | Chisada S.-I.,Japan National Research Institute of Fisheries And Environment of Inland Sea | Shimizu K.,Kyushu University | Shimizu K.,The Chemo Sero Therapeutic Research Institute KAKETSUKEN | And 5 more authors.
FEMS Microbiology Letters | Year: 2013

Vibrios, distributed in marine and brackish environments, can cause vibriosis in fish and shellfish under appropriate conditions. Previously, we clarified by thin-layer chromatography (TLC) overlay assay that 35S-labeled Vibrio trachuri adhered to GM4 isolated from red sea bream intestine. However, whether GM4 actually functions on epithelial cells as an attachment site for vibrios still remains to be uncovered. We found that six isolates, classified as V. harveyi, V. campbellii, and V. splendidus, from intestinal microflora of red sea bream adhered to GM4 but not galactosylceramide (GalCer) by TLC-overlay assay. Tissue-overlay assays revealed that V. harveyi labeled with green fluorescent protein (GFP) adhered to epithelial cells of red sea bream intestine where GM4 and GalCer were found to be distributed on the top layer of actin filaments by immunohistochemical analysis using corresponding antibodies. The number of adhering vibrios was diminished by pretreatment with anti-GM4 antibody, but not anti-GalCer antibody. These results clearly indicate that vibrios adhere to epithelial cells of red sea bream intestine utilizing GM4 as an attachment site. © 2013 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd.

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