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Bioethanol Production from Molasses by Yeasts with Stress-Tolerance Isolated from Aquatic Environments in Japan

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Advances in Microbiology 2019年第12期9卷 1000-1011页

作者： Yuka Naito Masahiko Okai masami ishida Masachika Takashio Naoto Urano Laboratory of Marine Biochemistry Department of Ocean Sciences Tokyo University of Marine Science and Technology Konan Minato-ku Tokyo Japan Zensho Laboratories of Food Technology Zensho Holdings Co. Ltd. Konan Minato-ku Tokyo Japan

Bioethanol is a safe and renewable source of energy that continues to be a research focus, since fossil fuels have been linked to global warming and nuclear energy sources are affected by the increased safety concerns following the 2011 nuclear power plant accident in Japan. In general, bioethanol is converted from a biomass by yeast fermentation. The production efficiency of this bioethanol is not sufficiently high, and its practical use as a substitute for fossil fuels and nuclear energy is thus limited. For the industrial production of bioethanol, the yeast fermentation of biomass cultures containing high concentration sugar, NaCl, and ethanol is necessary, but this might induce phenomena in which the stresses arising in the yeasts weaken their cells during fermentation. As described herein, we isolated 1028 strains of yeasts from natural aquatic environments: Japan’s Tama River and Lake Kasumigaura. Among them, 412 strains were fermentative yeasts and 31 strains showed high fermentation ability under a 30% sorbitol + 10% ethanol condition. These strains were identified as Torulaspola delbrueckii, Wickerhamomyces anomalus, Candida glabrata, Pichia kudriavzevii, Saccharomyces cf. cerevisiae/paradoxus, and Lachancea kluyveri. The strains T. delbrueckii, W. anomalus, and C. glabrata also showed tolerance against 15% NaCl. Most importantly, S. cf. cerevisiae/paradoxus H28 and L. kluyveri F2-67 produced 57.4 g/L and 53.9 g/L ethanol from molasses (sucrose 104.0 g/L, fructose 33.4 g/L, and glucose 24.8 g/L) within 48 hrs at 25°C, respectively.

关键词： Stress-Tolerance Molasses Yeast Species Aquatic Environment Fermentation

Characterization of Yeasts Capable of Neutralizing Acidic Media from Natural Neutral Environments

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Advances in Microbiology 2017年第12期7卷 887-897页

作者： Shintaro Nagaoka Takako Kobayashi Yuko Kajiwara Masahiko Okai masami ishida Naoto Urano Laboratory of Marine Biochemistry Department of Ocean Sciences Tokyo University of Marine Science and Technology Tokyo Japan

Acid-tolerant yeasts often inhabit extremely acidic environments: mine drain-ages, hot springs, and even fermented foods. Some of them also possess the ability to neutralize acidic media. However, the examples of these yeasts that are already known were isolated from acidic environments. In this study, the isolation of acid-tolerant yeasts from natural neutral aquatic environments and the identification of yeasts able to neutralize an acidic medium (acid-neutralizing yeast) in Japan were carried out. Various kinds of acid-tolerant and acid-neutralizing yeasts were obtained. In a neutralizing test using an acidic casamino acid solution adjusted to a pH of 4.0 with sulfuric acid, the obtained acid-neutralizing yeasts elevated the pH to approximately 7.0, and their neutralizing abilities were similar to those of previously reported yeasts that had been isolated from acidic environments. These results showed that acid-tolerant yeasts and acid-neutralizing yeasts exist widely in neutral environments, and little difference was found in the neutralizing abilities of yeasts obtained from neutral environments in comparison to those obtained from acidic environments.

关键词： Acid-Tolerant Yeast Acidic Environment Hydrosphere Neutralization Yeast Species

Molecular Phylogeny and Phenotypic Characterization of Yeasts with a Broad Range of pH Tolerance Isolated from Natural Aquatic Environments

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Advances in Microbiology 2019年第1期9卷 56-73页

作者： Naoto Urano Ayaka Shirao Yuka Naito Masahiko Okai masami ishida Masachika Takashio Department of Ocean Sciences Laboratory of Marine Biochemistry Tokyo University of Marine Science and Technology Tokyo Japan Zensho Laboratories of Food Technology Zensho Holdings Co. Ltd. Tokyo Japan

In this study, yeasts with broad range of pH tolerance were isolated and characterized from natural aquatic environments in Japan. Only a few basic and application studies of alkali-tolerant yeasts have been reported, despite the unmet industrial needs. First, we surveyed alkali-tolerant yeasts from natural aquatic environments at pH 7.6 - 9.4. We isolated 35 yeast strains that grew in pH 9.0 medium, from seven genera and nine species: 25 strains (N1, N2, through N6, N9, K1, K3 through K19) were Rhodotorula mucilaginosa;one (N7) was Rhodosporidium fluvial;one (N8) was Scheffersomyces spartinae;two (N10 and N13) were Wicherhamonyces anomalus;one (N11) was Cyberlindnera saturnus;one (S1) was Candida sp.;two (S2 and S4) were Candida intermedia;one (S3) was Candida quercuum;and one (K2) was Cryptococcus liquefacience. We examined the effects of pH on the growth of representative yeast strains. Strains K12 and S4 showed high growth at pH 3 - 10. Strains N7, N8, N10, N11, and S3 showed high growth at pH 3 - 9. Strains K2 and S1 showed high growth at pH 4 - 8. All nine of these strains had neutralizing activities from acidic media at pH 3 - 5 to pH 6 - 8. We previously isolated acid-tolerant yeasts (Cryptococcus sp. T1 [1] and Candida intermedia CeA16 [2]) from extremely acidified environments;they showed high growth at pH 3 - 9 and neutralizing activities of acidic media by releasing ammonium ions. Thus, alkali-tolerant yeasts and acid-tolerant yeasts were found to be similar species and have both high growth at a broad range of pH and neutralizing activities of acid media. Previously, we also isolated acid-tolerant, acid-neutralizing yeasts from neutral natural environments (26 strains, 12 species) [3]. Next, we constructed the phylogenetic trees of both acid-tolerant yeasts and alkali-tolerant yeasts. All were situated in the same classification position. Similar yeast species with a broad range of pH tolerance were living in natural aquatic environments at pH values fr

关键词： pH Tolerance Yeast Species Aquatic Environment Phylogenetic Tree Fermentation

Effect of Acid Species on <i>Cryptococcus</i>sp. T1 and Construction of Its Bioreactor System for Continuous Neutralization of Acidic Media

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Advances in Microbiology 2019年第1期9卷 74-86页

作者： Shintaro Nagaoka Yuko Yuko Kajiwara Sayaka Toyama Masahiko Okai masami ishida Naoto Urano Department of Ocean Science Laboratory of Marine Biochemistry Tokyo University of Marine Science and Technology Tokyo Japan Department of Life Science and Technology Tokyo Institute of Technology Yokohama Japan

Generally, the tolerance of yeasts to acid is stronger than that of bacteria. In addition, some of the yeasts are able to neutralize acidic media, which is called acid-neutralizing yeast. An acid-neutralizing yeast, Cryptococcus sp. strain T1, was previously isolated from Lake Tazawa in Japan. Here we investigated the effect of several types of acid on the neutralizing ability of strain T1, and we improved an existing bioreactor model to effectively neutralize acidic water. First we carried out a neutralizing test with strain T1 using casamino acid solution adjusted to pH 3.0 - 5.0 with each inorganic or organic acid. The solutions adjusted to pH 3.0 with nitric acid, phosphoric acid, sulfuric acid, and citric acid were neutralized by strain T1. The pH 3.0 solutions with lactic acid, formic acid, and acetic acid were not neutralized. These results demonstrated that some types of organic acid avoid neutralization by strain T1, and we hypothesized that this might be due to the sterilizing function of non-dissociative organic acid. We then constructed a bioreactor system with a column filled with T1-immobilized alginate beads. The previous research identified a problem in this bioreactor system: the high level of ammonium ions (NH+4) in the neutralized water might pollute water environments, and aluminum ions (Al3+) included in acidic water prevent the neutralization. Here, we used zeolite to get rid of the NH+4 and Al3+, and the addition of a zeolite reactor enabled the bioreactor system to neutralize the acidic water with a decrease of NH+4 in the water.

关键词： Acid-Tolerant Yeast Acid-Neutralizing Yeast Bioreactor Organic Acid Zeolite

Ethanol Fermentation by High-Stress-Tolerance Aquatic Yeasts and Their Mutants

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Advances in Microbiology 2021年第11期11卷 616-629页

作者： Naoto Urano masami ishida Yuka Naito Rintaro Endo Toshinori Takei Masachika Takashio Masahiko Okai Laboratory of Marine Biochemistry Department of Ocean Sciences Tokyo University of Marine Science and Technology Tokyo Japan Zensho Laboratories of Food Technology Zensho Holdings Co. Ltd. Tokyo Japan

Bioethanol is thought to be a renewable source of energy, because the biomasses used to make ethanol, such as sugar cane and its residual substance, molasses, are resources that can be continuously produced. But the practical use of ethanol to replace fossil fuels or atomic energy has been limited, because the production efficiencies of ethanol in relation to its substrates are not so high. Thus, for industrial production of the bioethanol, yeast fermentation would ideally be carried out in biomasses containing more highly concentrated carbohydrates. However, the environmental stresses in highly concentrated cultures might weaken the yeast’s physiological activities. From various kinds of aquatic yeast with stress tolerance, Torulaspora derbrueckii F2-11 and Wicherhamomyces anomalus AN2-64 were selected as candidates for high-sugar-tolerance yeasts as they showed remarkable growth in the YPD + sorbitol (600 g/L) medium at 25°C for 120 hrs. When the amounts and kinds of sugar alcohols in the cells of the two strains were measured in cultures containing 20 g/L or 400 g/L of D-glucose, maltose, or sucrose, the main two sugar alcohols that accumulated as the sugar concentration increased were glycerol and arabitol. Mutation by ethyl methanesulfonate of the parent strains T. derbrueckii F2-11 and W. anomalus AN2-64 induced mutants F2-11M or AN2-64M, which showed higher sugar, heat, and ethanol tolerances than their respective parents. Ethanol productivities and sugar assimilation activities of the mutants were also higher than those of the parents in the 25% (v/v) molasses.

关键词： Ethanol Fermentation Stress Tolerance Aquatic Yeast Mutant Ethyl

Behavior of Antibiotic-Resistant Fecal Coliforms in the Stream of a Sewage Treatment Plant in Tokyo

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Advances in Microbiology 2020年第7期10卷 318-330页

作者： Naoto Urano Masahiko Okai Yusuke Tashiro Azusa Takeuchi Rintaro Endo masami ishida Masachika Takashio Laboratory of Marine Biochemistry Department of Ocean Sciences Tokyo University of Marine Science and Technology Tokyo Japan Zensho Laboratories of Food Technology Zensho Holdings Co. Ltd. Tokyo Japan

We are confronting a new threat in the prevalence of antibiotic-resistant bacteria followed by epidemic spread in aquatic environments in metropolitan areas because damage from river floods is increasing remarkably in Japan due to global extreme weather. The sewer penetration rate is about 100% in Tokyo and reclaimed water from sewage treatment plants accounts for over 50% of all water in both the down- and mid-stream areas of local rivers. The water quality of these rivers, which contain microflora, seems to be seriously affected by reclaimed water. In this study, we collected water samples on July 17, 2018 and examined the behavior of antibiotic-resistant fecal coliforms in the stream of a sewage treatment plant in Tokyo. Extended-spectrum β-lactamase (ESBL)-producing fecal coliforms with encoding genes were found;the CTX-M-1, CTX-M-9, TEM, and SHV groups were found to have survived in the final effluent to the river after sterilization with sodium hypochlorite.

关键词： Antibiotic-Resistant Bacteria (ARB) Extended-Spectrum β-Lactamase (ESBL) Fecal Coliforms Sewage Treatment Plant Reclaimed Water

Effects of Fuscoporia obliqua on Postprandial Glucose Excursion and Endothelial Dysfunction in Type 2 Diabetic Patients

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Journal of Traditional Chinese Medicine 2008年第1期28卷 49-57页

作者： Toshihiro Maenaka masami Oshima Yuka Itokawa Takashi Masubuchi Yasuyuki Takagi Jung-Sook Choi Torao ishida Yeunhwa Gu Graduate School of Health Science Suzuka University of Medical ScienceJapan Gyeongdo provincial college Kyoungbook Korea Hi-tech Research Center Suzuka University of Medical Science Japan Graduate School of Health Science Suzuka University of Medical Science Japan Hi-tech Research Center Suzuka University of Medical Science Japan

Postprandial hyperglycemia has been reported to elicit endothelial dysfunction and provoke future cardiovascular complications. A reduction of postprandial blood glucose levels by the glucosidase inhibitor Fuscoporia obliqua was associated with a risk reduction of cardiovascular complications, but the effects of Fuscoporia obliqua on endothelial function have never been elucidated. This study is aimed to assess the efficacy of Fuscoporia obliqua on postprandial metabolic parameters and endothelial function in type 2 diabetic patients. Postprandial peak glucose (14.47±1.27 vs. 8.50±0.53 mmol/liter), plasma glucose excursion (PPGE), and change in the area under the curve (AUC) glucose after a single loading of test meal (total 450 kcal; protein 15.3%; fat 32.3%; carbohydrate 51.4%) were significantly higher in the diet-treated type 2 diabetic patients (n=14) than the age-and sex-matched controls (n=12). The peak forearm blood flow response and total reactive hyperemic flow (flow debt repayment) during reactive hyperemia, indices of resistance artery endothelial function on strain-gauge plethysmography, were unchanged before and after meal loading in the controls. But those of the diabetics were significantly decreased 120 and 240 min after the test meal. A prior administration of Fuscoporia obliqua decreased postprandial peak glucose, PPGE, and AUC glucose. The peak forearm blood flow and flow debt repayment were inversely well correlated with peak glucose, PPGE, and AUC glucose, but not with AUC insulin or the other lipid parameters. Even a single loading of the test meal was shown to impair the endothelial function in type 2 diabetic patients, and the postprandial endothelial dysfunction was improved by a prior use of Fuscoporia obliqua. Fuscoporia obliqua might reduce macrovascular complication by avoiding endothelial injury in postprandial hyperglycemic status.

关键词： Fuscoporia oblique Glucose excursion and Endothelial Dysfunction Type 2 Diabetic Patients Insulin

Effects of K. Lysolecithin on Blood Levels of Monoamines in Mice

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Journal of Traditional Chinese Medicine 2007年第3期27卷 212-219页

作者： Yuka Itokwa Aki Kozu Takashi Nakamura Takashi Masubuchi masami Oshima Kwang-Ho Cho Jung-Sook Choi Toshihiro Miura Torao ishida Yeunhwa Gu Suzuka Univercity of Medical Science Graduate School of Health Science Society of Preventive and Alternative Medicine Department of Radiological Science Catholic University of Deagu College of Health Science Gyeongdo Provincial College

In this research, Lysolecithin-a substance made with 100% natural ingredients - was given to ICR mice as medication to measure its periodic effect on the noradrenalin (NA), dopamine (DA), and serotonin (5-HT) levels of the brain. Both ICR and SAM mice were separated into two groups - control group and Lysolecithin (K. Lysolecithin: hydrolytic lysolecithin) medicated group, and given 1-week preparation period. The K. Lysolecithin group was given 500mg/kg of K. Lysolecithin at 0.2mL per dosage for 4 weeks, and the control group was given the same amount of dosage of water during the same period. NA, DA and 5-HT concentrations were measured from the blood before medication and 8 weeks / 12 weeks / 16 weeks after the first medication. For the SAM mice, 8 weeks after they were medicated with K .Lysolecithin, Morris Water Maze Test was conducted for 7 consecutive days and then the concentrations were measured by drawing blood from the heart. The K. Lysolecithin medicated group showed a tendency to have a statistically significant higher concentrations of 5-HT and NA in the blood. Also, periodic examination showed that the monoamine levels were highest in the 12th week and declined thereafter.

关键词： K. Lysolecithin Serotonin Noradrenalin Dopamine Alzheimer's Syndrome