Expression and characterization of a bifunctional thermal β-glucosidase IuBgl3 from thermophilic archaeon Infirmifilum uzonense / 生物工程学报

β-glucosidase has important applications in food, medicine, biomass conversion and other fields. Therefore, exploring β-glucosidase with strong stability and excellent properties is a research hotspot. In this study, a GH3 family β-glucosidase gene named Iubgl3 was successfully cloned from Infirmifilum uzonense. Sequence analysis showed that the full length of Iubgl3 was 2 106 bp, encoding 702 amino acids, with a theoretical molecular weight of 77.0 kDa. The gene was cloned and expressed in E. coli and the enzymatic properties of purified IuBgl3 were studied. The results showed that the optimal pH and temperature for pNPG hydrolysis were 5.0 and 85 ℃, respectively. The enzyme has good thermal stability, and more than 85% of enzyme activity can be retained after being treated at 80 ℃ for2 h. This enzyme has good pH stability and more than 85% of its activity can be retained after being treated at pH 4.0-11.0 for 1 h. It was found that the enzyme had high hydrolysis ability to p-nitrophenyl β-d-glucoside (pNPG) and p-nitrophenyl β-d-xylopyranoside (pNPX). When pNPG was used as the substrate, the kinetic parameters Km and Vmax were 0.38 mmol and 248.55 μmol/(mg·min), respectively, and the catalytic efficiency kcat/Km was 6 149.20 s-1mmol-1. Most metal ions had no significant effect on the enzyme activity of IuBgl3. SDS completely inactivated the enzyme, while EDTA increased the enzyme activity by 30%. This study expanded the β-glucosidase gene diversity of the thermophilic archaea GH3 family and obtained a thermostable acid bifunctional enzyme with good industrial application potential.

Effect of thermophilic bacterium HB27 manganese superoxide dismutase in a rat model of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) / 亚洲男科学杂志(英文版)

We investigated the therapeutic effects of superoxide dismutase (SOD) from thermophilic bacterium HB27 on chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and its underlying mechanisms. A Sprague-Dawley rat model of CP/CPPS was prepared and then administered saline or Thermus thermophilic (Tt)-SOD intragastrically for 4 weeks. Prostate inflammation and fibrosis were analyzed by hematoxylin and eosin staining, and Masson staining. Alanine transaminase (ALT), aspartate transaminase (AST), serum creatinine (CR), and blood urea nitrogen (BUN) levels were assayed for all animals. Enzyme-linked immunosorbent assays (ELISA) were performed to analyze serum cytokine concentrations and tissue levels of malondialdehyde, nitric oxide, SOD, catalase, and glutathione peroxidase. Reactive oxygen species levels were detected using dichlorofluorescein diacetate. The messenger ribonucleic acid (mRNA) expression of tissue cytokines was analyzed by reverse transcription polymerase chain reaction (RT-PCR), and infiltrating inflammatory cells were examined using immunohistochemistry. Nuclear factor-κB (NF-κB) P65, P38, and inhibitor of nuclear factor-κBα (I-κBα) protein levels were determined using western blot. Tt-SOD significantly improved histopathological changes in CP/CPPS, reduced inflammatory cell infiltration and fibrosis, increased pain threshold, and reduced the prostate index. Tt-SOD treatment showed no significant effect on ALT, AST, CR, or BUN levels. Furthermore, Tt-SOD reduced inflammatory cytokine expression in prostate tissue and increased antioxidant capacity. This anti-inflammatory activity correlated with decreases in the abundance of cluster of differentiation 3 (CD3), cluster of differentiation 45 (CD45), and macrophage inflammatory protein 1α (MIP1α) cells. Tt-SOD alleviated inflammation and oxidative stress by reducing NF-κB P65 and P38 protein levels and increasing I-κBα protein levels. These findings support Tt-SOD as a potential drug for CP/CPPS.

Statistical optimization of cellulases by Talaromyces thermophilus utilizing Saccharum spontaneum, a novel substrate

BACKGROUND: At present, cellulases are the most important enzymes worldwide, and their demand has been increasing in the industrial sector owing to their notable hydrolysis capability. RESULTS: In the present study, contrary to conventional techniques, three physical parameters were statistically optimized for the production of cellulase by thermophilic fungi by using response surface methodology (RSM). Among all the tested thermophilic strains, the best cellulase producing fungus was identified as Talaromyces thermophilus ­ both morphologically and molecularly through 5.8S/ITS rDNA sequencing. The central composite design (CCD) was used to evaluate the interactive effect of the significant factors. The CCD was applied by considering incubation period, pH, and temperature as the model factors for the present investigation. A second-order quadratic model and response surface method revealed that the independent variables including pH 6, temperature 50 C, and incubation period 72 h significantly influenced the production of cellulases. The analysis of variance (ANOVA) indicated that the established model was significant (P 0.05) and showed the high adequacy of the model. The actual and predicted values of CMCase and FPase activity showed good agreement with each other and also confirmed the validity of the designed model. CONCLUSIONS: We believe the present findings to be the first report on cellulase production by exploiting Kans grass (Saccharum spontaneum) as a substrate through response surface methodology by using thermophilic fungus, Talaromyces thermophilus.

Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates

Abstract Lipases are currently used in food technology for the modification of fats and oils. The thermal stability of lipase is an essential characteristic for this application. This study compares four individual single-step methods (heat treatment, ethanol precipitation, ammonium sulfate precipitation, and size-exclusion chromatography) to enrich lipase concentrations from thermophilic bacterial (Geobacillius stearothermophilus and Anoxybacillus flavithermus) cell lysates. SDS-PAGE and size exclusion chromatography were used to determine the molecular weights of the lipases and the enrichment efficiencies were determined using specific enzyme activities. The molecular weight of G. stearothermophilus lipase was approximately 42 kDa, and approximately 33 kDa for A. flavithermus lipase. For each organism, ethanol precipitation resulted in the highest enrichment fold, followed by ammonium sulfate precipitation, gel filtration and heat treatment respectively. The highest yields for G. stearothermophilus lipase were obtained with ammonium sulfate precipitation, followed by get filtration, and ethanol precipitation respectively. The highest yields for A. flavithermus lipase were obtained from heat treatment followed by ammonium sulfate precipitation, gel filtration and ethanol precipitation respectively. Ethanol precipitation and heat treatment are simple methods for enzyme enrichment from cell lysates and can result in high enzyme yields with moderate enrichment folds compared to complex multi-step purification methods.

Purification and biochemical characterization of a novel thermophilic exo-ß-1, 3-glucanase from the thermophile biomass-degrading fungus Thielavia terrestris Co3Bag1

Background: The aim of this work was to purify and characterize exo-ß-1,3-glucanase, namely, TtBgnA, from the thermophilic fungus Thielavia terrestris Co3Bag1 and to identify the purified enzyme. Results: The thermophilic biomass-degrading fungus T. terrestris Co3Bag1 displayed ß-1,3-glucanase activity when grown on 1% glucose. An exo-ß-1,3-glucanase, with an estimated molecular mass of 129 kDa, named TtBgnA, was purified from culture filtrates from T. terrestris Co3Bag1. The enzyme exhibited optimum activity at pH 6.0 and 70°C and half-lives (t1/2) of 54 and 37 min at 50 and 60°C, respectively. Substrate specificity analysis showed that laminarin was the best substrate studied for TtBgnA. When laminarin was used as the substrate, the apparent KM and Vmax values were determined to be 2.2 mg mL-1 and 10.8 U/mg, respectively. Analysis of hydrolysis products by thin-layer chromatography (TLC) revealed that TtBgnA displays an exo mode of action. Additionally, the enzyme was partially sequenced by tandem mass spectrometry (MS/MS), and the results suggested that TtBgnA from T. terrestris Co3Bag1 could be classified as a member of the GH-31 family. Conclusions: This report thus describes the purification and characterization of TtBgnA, a novel exo-ß-1,3-glucanase of the GH-31 family from the thermophilic fungus T. terrestris Co3Bag1. Based on the biochemical properties displayed by TtBgnA, the enzyme could be considered as a candidate for potential biotechnological applications.

Identificação de micro-organismos presentes em sistemas de biolixiviação de cobre a 35 e 50°C / Identifying microorganisms related to copper bioleaching at 35 and 50°C

RESUMO A biolixiviação de minérios de baixo teor e com elevado conteúdo de impurezas tem se mostrado alternativa importante para o aproveitamento destes, uma vez que a recuperação do metal por métodos pirometalúrgicos convencionais mostra-se economicamente inviável. A identificação e quantificação dos micro-organismos capazes de promover a biolixiviação mostram-se estratégicas para alcançar bons rendimentos no controle do processo e na recuperação de metais. Nesse sentido, as técnicas de biologia molecular são as ferramentas mais utilizadas para tal propósito. Este trabalho, utilizando técnicas de reação em cadeia da polimerase (PCR), polimorfismos de comprimento dos fragmentos de restrição (RFLP) e reação em cadeia da polimerase seguida de eletroforese em gel com gradiente desnaturante (PCR-DGGE), mostrou que a diversidade nas colunas de biolixiviação de cobre estudadas é baixa e que a temperatura é importante na manutenção de determinadas espécies, havendo predominância de Acidithiobacillus ferroxidans a 35°C e de Sulfobacillus thermosulfidooxidans a 50°C.

ABSTRACT Bioleaching is an alternative to pyrometallurgy for the production of metals from low-grade ores containing high level of impurities, once that live pyrometallurgical methods are economically unfeasible. The quantification and identification of those microorganisms related to bioleaching is an important strategy for process control and thus metal recovery. In this regard, molecular biology is one of the main techniques utilized for such objective. This study applied PCR, RFLP and PCR-DGGE techniques to show that the microbial diversity in copper bioleaching columns under investigation is low and the temperature is important to define the species found, with predominance of Acidithiobacillus ferroxidans, at 35°C and Sulfobacillus thermosulfidooxidans at 50°C.

A new isolate of thermophilic and organic solvent tolerant bacteria for lipase production using basal medium of palm kernel cake 80-87

@#Aims: This research focused on the selection of potential strains especially bacteria that can grow effectively in palm kernel cake (PKC) and produce high amount of thermostable and solvent tolerant (TS-OST) lipase. The work involved the exploration of renewable PKC as potential fermentation medium for discovery to novel TS-OST lipase that would have excellent tolerance and activity in presence of organic solvents with high temperatures for industrial applications.Methodology and results: Using palm kernel cake (PKC) as source of thermophilic bacteria, 53 bacterial strains were found survived at temperature 65 °C. However, after subcultured several times, only 17 strains were found as pure thermophilic strains. Preliminary screening both qualitative and quantitative was performed to all 17 potential thermophilic bacterial strains and showed that only 11 purified thermophilic strains are lipase producer. Strain PKC-P1 produced highest enzyme activity (11.13 U/g), followed by PKC-P13 and PKC-C9. The lowest enzymeactivity was lipase produced byPKC-C10 (0.76U/g). Strain PKC-P1 has been classified as Gram negative bacteria and identified as Bacillus smithiistrain PKC_P1.Conclusion, significance and impact of study: PKC as a by-product of oil palm industry consistsof many nutrients that can give benefits towards industry and can be utilized in order to produce enzymes like lipases. From these results, it could be concluded that this lipase stable at temperature 65 °C and pH 7 and may be a potential candidate to be used in a variety of biotechnological applications. This finding revealed that a bacterial strain obtained from oil-rich environment which is PKC through isolation process has potential as a source of more economical enzyme to be applied in biotechnology industr

Analysis of Microbial Contamination Status of 9 Kinds of Chinese Herbal Pieces from Guangxi Regions / 中国药师

Objective:To investigate the microbial contamination status of Chinese herbal pieces from Guangxi regions and provide reference for the microbial limit standard. Methods:According to Chinese Pharmacopoeia (2015 edition), the herbal pieces were analyzed for the total number of aerobe microbial, yeasts, molds, thermophilic bacteria and bile-resistant gram-negative bacteria,and salmonella was also detected. Results:In 90 batches of Chinese herbal pieces,the lg average value of the total number of aerobe microbial was 3.7,and that of the total yeasts and molds was 1.7,that of thermophilic bacteria was 0.9 and that of bile-resistant gram-negative bacteria was 1.1,and Salmonella was not detected out. There were statistically significant differences (P < 0.05) in the microbial contamination levels of herbal pieces with different processing techniques and packaging materials. Conclusion:According to different processing methods and administration, microbial limit standard should be established for classification in order to guide the standardized production of Chinese herbal pieces and improve their quality gradually.

Effects of volatile fatty acids in biohydrogen effluent on biohythane production from palm oil mill effluent under thermophilic condition

Background: Biohydrogen effluent contains a high concentration of volatile fatty acid (VFA) mainly as butyric, acetic, lactic and propionic acids. The presence of various VFAs (mixture VFAs) and their cooperative effects on two-stage biohythane production need to be further studied. The effect of VFA concentrations in biohydrogen effluent of palm oil mill effluent (POME) on methane yield in methane stage of biohythane production was investigated. Results: The methane yield obtained in low VFA loading (0.9 and 1.8 g/L) was 15­20% times greater than that of high VFA loading (3.6 and 4.7 g/L). Butyric acid at high concentrations (8 g/L) has the individual significantly negative effect the methane production process (P b 0.05). Lactic, acetic and butyric acid mixed with propionic acid at a concentration higher than 0.5 g/L has an interaction significantly negative effect on the methanogenesis process (P b 0.05). Inhibition condition had a negative effect on both bacteria and archaea with inhibited on Geobacillus sp., Thermoanaerobacterium thermosaccharolyticum, Methanoculleus thermophilus and Methanothermobacter delfuvii resulting in low methane yield. Conclusion: Preventing the high concentration of butyric acid, and propionic acid in the hydrogenic effluent could enhance methane production in two-stage anaerobic digestion for biohythane production.

Investigation the Consumption of Aromatic Hydrocarbons by Anoxybacillus rupeinsis Ir3 (JQ912241) using FTIR and HPLC Analyses.

Aims: To confirm the ability of Anoxybacillus rupiensis strain Ir3 (JQ912241) to utilize the aromatic compounds using FTIR and HPLC analyses. Study Design: Experimental study. Place and Duration of Study: Department of Biotechnology, College of Science, Al-Nahrain University. Baghdad, Iraq, between December 2012 and April 2013. Methodology: Anoxybacillus rupiensis strain Ir3 (JQ912241), a newly thermophilic bacterium, isolated from hydrocarbon contaminated soil in Iraq, was used. Analytical experiments include HPLC (High performance liquid chromatography) and FTIR (Fourier transform infrared) were used to determine the ability of this strain to utilize the aromatic compounds. Results: The quantitative analysis (HPLC) indicated that this bacterium showed as much as 99.62% consumption of carbazole, 99.4% of ρ-nitrophenole, 97.73% of nitrobenzene and 98.89% of naphthalene. Qualitative analysis of FTIR spectra showed that A. rupiensis strain Ir3 (JQ912241) has the ability to convert carbazole to anthranilic acid, indicating the presence of the meta cleavage enzyme, this also confirmed by using 2, 3-dihydroxybiphenyl through converting the colony color on Luria-Bertani (LB) and minimal agar plates to brown. Conclusion: The good ability of A. rupiensis strain Ir3 to utilize the studied aromatic hydrocarbons make it a good candidate as biocatalist. Its ability to convert carbazole to anthranilic acid and to oxidize catechol to 2-hydroxymuconic semialdehyde is through the meta cleavage enzyme.

Evaluation of biological production of lactic acid in a synthetic medium and in Aloe vera (L.) Burm. f. processing by-products / Evaluation of biological production of lactic acid in a synthetic medium and in Aloe vera (L.) Burm. f. processing by-products / Avaliação da produção biológica de ácido lático em um meio sintético e no processamento de subprodutos de Aloe vera (L.) Burm. f.

This study evaluated lactic acid production through batch fermentation in a bioreactor with Thermoanaerobacter sp. strain USBA-018 and a chemically defined culture medium and with hydrolyzed pressed extract of Aloe vera peel (AHE). The strain USBA-018 fermented various sugars, but its primary end-product was L-lactic acid. Factors which influenced L- lactic acid production were pH, addition of yeast extract (YE) and manganese chloride. Under the most favorable growing conditions for the production of lactic acid, yield (Yp/s) increased from 0.66 to 0.96 g/g with a productivity (Qp) of 0.62 g.l-1.h and a maximum lactic acid concentration of 178 mM at 26 hours of fermentation. When AHE was used, 93.3 mM, or 0.175 g.l-1.h, was obtained. These results show the potential for transformation of sugars that strain USBA-018 offers, but additional studies are needed to find out if different strategies using AHE as carbon source can produce large enough quantities of lactic acid to allow AHE to become a low-cost alternative substrate.

Este estudio evaluó la producción de ácido láctico de la cepa de Thermoanaerobacter sp. USBA-018 en un biorreactor de fermentación por lotes, utilizando como medios de cultivo una formulación químicamente definida y un extracto prensado e hidrolizado de cáscara de Aloe vera (AHE). La cepa USBA-018 fermentó varios azúcares, pero su principal producto final fue L-ácido láctico. Los factores que influyeron en la producción de L-ácido láctico fueron pH, adición de extracto de levadura (YE) y de cloruro de manganeso. Bajo las condiciones más favorables de crecimiento para la producción de ácido láctico el rendimiento (Yp/s) aumentó de 0.66 a 0.96 g/g, con una productividad (Qp) de 0.62 g.l-1.h y una máxima concentración de ácido láctico de 178 mM a las 26 horas de fermentación. Cuando se usó AHE, se obtuvieron 93.3 mM, o 0.175 g.l-1.h. Estos resultados muestran el potencial de transformación de azúcares que ofrece la cepa USBA-018, pero se requieren estudios adicionales para determinar si diferentes estrategias de uso de AHE como fuente de carbono producen cantidades suficientemente grandes de ácido láctico como para permitir que el AHE se convierta en un sustrato alternativo de bajo costo.

Este estudo avaliou a produção de ácido lático por meio de fermentação descontinua em um biorreator com Thermoanaerobacter sp. cepa USBA-018 e um meio de cultura químicamente definido e com extrato hidrolisado de casca de Aloe vera (AHE). A cepa USBA-018 fermentou vários acucares, mas seu produto final primàrio foi L-ácido lático. Os fatores que influenciaram a produção de L-ácido lático foram o pH, adição de extrato de levedura (YE) e cloreto de manganès. Sob as condicoes de crescimento mais favoráveis para a produção de ácido lático, o rendimento (Yp/S) aumentou de 0,66 a 0,96 g/g com uma produtividade (Q) de 0,62 g.L4.h e um máximo de concentração de ácido lático de 178 mM em 26 horas de fermentação. Quando o AHE foi utilizado, se obteve 93,3 mM ou 0,175 g.L-1.h. Estes resultados mostram o potencial para transformação de acucares que a cepa USBA-018 oferece, entretanto estudos adicionais sáo necessários para descobrir se diferentes estrategias utilizando o AHE como fonte de carbonos podem produzir quantidades de ácido lático grandes o suficiente para permitir que AHE se converta em um substrato alternativo de baixo custo.

Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils

The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. subtilis (subsp. spizizenii (2) and subsp. inaquosurum (6)), B. amyloliquefaciens (subsp. amyloliquefaciens (4) and subsp. plantarum (4)), B. tequilensis (3), B. pumilus (3) and Bacillus sp. (19). Only six isolates (2.5%) belonged to the genus Aeribacillus represented by A. pallidus (4) and Aeribacillus sp. (2). In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively.

Screening of thermotolerant and thermophilic fungi aiming β-xylosidase and arabinanase production

Plant cell wall is mainly composed by cellulose, hemicellulose and lignin. The heterogeneous structure and composition of the hemicellulose are key impediments to its depolymerization and subsequent use in fermentation processes. Thus, this study aimed to perform a screening of thermophilic and thermotolerant filamentous fungi collected from different regions of the São Paulo state, and analyze the production of β-xylosidase and arabinanase at different temperatures. These enzymes are important to cell wall degradation and synthesis of end products as xylose and arabinose, respectively, which are significant sugars to fermentation and ethanol production. A total of 12 fungal species were analyzed and 9 of them grew at 45 ºC, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 ºC. Aspergillus niger and Aspergillus thermomutatus were the filamentous fungi with the most expressive production of β-xylosidase and arabinanase, respectively. In general for most of the tested microorganisms, β-xylosidase and arabinanase activities from mycelial extract (intracellular form) were higher in cultures grown at high temperatures (35-40 ºC), while the correspondent extracellular activities were favorably secreted from cultures at 30 ºC. This study contributes to catalogue isolated fungi of the state of São Paulo, and these findings could be promising sources for thermophilic and thermotolerant microorganisms, which are industrially important due to their enzymes.

04-1 Fangotherapy, a body-friendly treatment to use hot spring water for health promotion / 日本温泉気候物理医学会雑誌

Fangotherapy is one of four medical treatments used under a medical doctor’s supervision at a hot spring. This treatment is conducted in Europe, especially in Italy, using peloids maturated with natural hot spring water. The maturated peloids have factors of concentrated hot spring and biological extract -glycolipids- from thermophilic algae.   There are many kinds of hot springs in Japan, but they are almost always used for “taking a bath” only. Our research focused on Fango found in Abano Italy, and modified into “Japanese style Fango” made with maturated peloids by hot spring water. Methods: The original Japanese Fango, which we made in a hot spring in Japan, has been named Biofango®. The original and first Biofango® was made from the Sanrakuen Hotel’s hot spring water in Toyama, and treated in the hotel as in the Abano style. The benefits of Biofango® were checked using the following methods; 1.　Hyperthermia and some medical effects were checked under treatment with maturated peloids, and the effects were compared with hot spring water only and with boiled tap water. 2.　The double-blind method was used comparing Biofango® (True Fango) and imitation Fango. Two kinds of Fango were made using either hot spring water or hot tap water, and some medical effects were checked. After treatment of Fangotherapy, a medical questionnaire was provided for each test subject. Results: Fango (Biofango®) is the best treatment among three bathing methods for keeping normal responses of blood flow and blood pressure at the thigh, and for keeping good thermal effects on the body, especially for the back of the body after 50 min.   The results of the double-blind method, deep body temperature increase and diastolic blood pressure decrease in True Fango showed a significant difference. Pulse increase and SIV decrease in True Fango shows a reduced load on the vessel, while still showing a high thermal effect. Functions of the body showed an improved friend in True Fango, according to the questionnaire survey. Conclusion: Traditional use of hot spring water in Japan was hot spring bathing in the mainstream. But, according to these results, Fango is the best method for a body-friendly treatment by hot spring water. Further, the questionnaire about body functions after Fango treatment shows that the hot spring Fango (True Fango) is more effective than hot tap water Fango (imitation Fango).   In Italy, glycolipids from thermophilic algae are an important factor for reducing inflammation by Fango therapy. Growth of thermophilic algae also has been confirmed in Fango maturation in Japan, and was cultured in the laboratory. We also found such glycolipids from Japanese algae, and have evidence, in situ, of the glycolipids remaining between particles of peloids. In the future, Biofango® should have a useful biological factor similar to Abano Fango for treatment.

04-1Fangotherapy, a body-friendly treatment to use hot spring water for health promotion / 日本温泉気候物理医学会雑誌

Fangotherapy is one of four medical treatments used under a medical doctor’s supervision at a hot spring. This treatment is conducted in Europe, especially in Italy, using peloids maturated with natural hot spring water. The maturated peloids have factors of concentrated hot spring and biological extract -glycolipids- from thermophilic algae. <BR>  There are many kinds of hot springs in Japan, but they are almost always used for “taking a bath” only. Our research focused on Fango found in Abano Italy, and modified into “Japanese style Fango” made with maturated peloids by hot spring water. <BR><b>Methods: </b>The original Japanese Fango, which we made in a hot spring in Japan, has been named Biofango^®. The original and first Biofango^® was made from the Sanrakuen Hotel’s hot spring water in Toyama, and treated in the hotel as in the Abano style. The benefits of Biofango^® were checked using the following methods;<BR>1.　Hyperthermia and some medical effects were checked under treatment with maturated peloids, and the effects were compared with hot spring water only and with boiled tap water. <BR>2.　The double-blind method was used comparing Biofango^® (True Fango) and imitation Fango. Two kinds of Fango were made using either hot spring water or hot tap water, and some medical effects were checked. After treatment of Fangotherapy, a medical questionnaire was provided for each test subject.<BR><b>Results: </b>Fango (Biofango^®) is the best treatment among three bathing methods for keeping normal responses of blood flow and blood pressure at the thigh, and for keeping good thermal effects on the body, especially for the back of the body after 50 min. <BR>  The results of the double-blind method, deep body temperature increase and diastolic blood pressure decrease in True Fango showed a significant difference. Pulse increase and SIV decrease in True Fango shows a reduced load on the vessel, while still showing a high thermal effect. Functions of the body showed an improved friend in True Fango, according to the questionnaire survey.<BR><b>Conclusion: </b>Traditional use of hot spring water in Japan was hot spring bathing in the mainstream. But, according to these results, Fango is the best method for a body-friendly treatment by hot spring water. Further, the questionnaire about body functions after Fango treatment shows that the hot spring Fango (True Fango) is more effective than hot tap water Fango (imitation Fango).<BR>  In Italy, glycolipids from thermophilic algae are an important factor for reducing inflammation by Fango therapy. Growth of thermophilic algae also has been confirmed in Fango maturation in Japan, and was cultured in the laboratory. We also found such glycolipids from Japanese algae, and have evidence, <i>in situ</i>, of the glycolipids remaining between particles of peloids. In the future, Biofango^® should have a useful biological factor similar to Abano Fango for treatment.

Isolation and Analysis of the Enzymatic Properties of Thermophilic Fungi from Compost

To the best of our knowledge, this is the first report on thermophilic fungi isolated in Korea. Three species of thermophiles were isolated from compost and were identified as Myriococcum thermophilum, Thermoascus aurantiacus, and Thermomyces lanuginosus. They can grow at temperatures above 50degrees C and produce high levels of cellulolytic and xylanolytic enzymes at high temperatures. Notably, the considerable thermostability of the endo-glucanase produced by T. aurantiacus has made the fungus an attractive source of industrial enzymes.

Isolation and characterization of novel thermophilic lipase-secreting bacteria

The purpose of the present study was to screen and identify the lipase-producing microorganisms from various regions of Iran. Samples collected from hot spring, Persian Gulf, desert area and oil-contaminated soil, were analyzed for thermophilic extracellular-lipase producing organisms. Six strains with high activity on rhodamine B plates were selected for chemical identification and further study. Among these isolated bacteria, four strains show higher activity in pH-Stat method at 55 °C. These strains were identified by PCR amplification of 16s rRNA genes using universal primers. Fermentation increased the activity up to 50%. The growth medium, designed for lipase production, increased the activity up to 4.55 folds. The crude supernatant of ZR-5 after fermentation and separation the cells, was lyophilized and the activity was measured. Total activity of this strain was 12 kU/g that shows its potential for industrial uses. Further study is required for purification of enzyme and calculation its specific activity. Immobilization is another approach should be considered.

Effects of agitation and exogenous H2 on bioconversion of sugarcane bagasse into ethanol by Clostridium thermocellum ATCC 27405

Background: The production of ethanol by a Consolidated Bioprocessing (CBP) strategy, which simultaneously combines cellulase production, lignocellulosic biomass hydrolysis and fermentation of released sugars to ethanol in one bioreactor, is a promising technology for cost reduction in the biological processing of biomass, specially using agroindustrial residues. Clostridium thermocellum is an anaerobic, thermophilic, strictly fermentative gram positive bacterium that meets all the requirements for CBP. Results: Ethanol concentration obtained in the non-stirred fermentation process in flasks with raw bagasse was two times greater than that in the stirred system. The results observed using a pretreated sugarcane bagasse in non-stirred flasks regarding ethanol concentration, were slightly lower than with raw bagasse. The sparging of exogenous H2 into the medium at atmospheric pressure inside the bioreactor showed to be unfavourable to achieve higher ethanol yields. Conclusions: The strain investigated is a promising candidate for thermophilic fermentative ethanol production from dried ground raw sugarcane bagasse in a CBP strategy, although the alcohol concentrations need to be further improved. In future studies, it is recommended to investigate different modes of operation of the fermentation process, including pressurized conditions, as well as to use wet raw sugarcane bagasse aiming to achieve additional improvement in ethanol production and to reduce the costs of the process.

Biohydrogen production by Thermoanaerobacterium thermosaccharolyticum KKU-ED1: Culture conditions optimization using mixed xylose/arabinose as substrate

Background: Biological hydrogen production by microorganisms can be divided into two main categories i.e. photosynthetic organisms that produce hydrogen using light as energy source and anaerobic bacteria that produce hydrogen via dark fermentation. Dark fermentative hydrogen production by anaerobic bacteria has the advantages of a higher HPR without illumination and of the capability to convert various kinds of substrate. Results: Thermophilic hydrogen producer was isolated from elephant dung and identified as Thermoanaerobacterium thermosaccharolyticum KKU-ED1 by 16S rRNA gene analysis, which was further used to produce hydrogen from mixed pentose sugar i.e., xylose/arabinose. The optimum conditions for hydrogen production from mixed xylose/arabinose by KKU-ED1 were a 1:1 xylose/arabinose mixture at the total concentration of 5 g/L, initial pH of 6.5 and temperature of 55ºC. Under the optimum conditions, hydrogen from sugar derived from acid-hydrolyzed sugarcane bagasse at a reducing sugar concentration were achieved. Soluble metabolite product (SMP) was predominantly acetic acid indicating the acetate-type fermentation. Conclusions: The strain KKU-ED1 appeared to be a suitable candidate for thermophilic fermentative hydrogen production from hemicellulosic fraction of lignocellulosic materials due to its ability to use various types of carbon sources.

Utilization of Agro-industrial Wastes for the Simultaneous Production of Amylase and Xylanase by Thermophilic Actinomycetes

Agro-industrial wastes such as sugarcane bagasse, wheat bran, rice bran, corn cob and wheat straw are cheapest and abundantly available natural carbon sources. The present study was aimed to production of amylase and xylanase simultaneously using agro-industrial waste as the sole carbon source. Seven thermophilic strains of actinomycete were isolated from the mushroom compost. Among of these, strain designated MSC702 having high potential to utilize agro-industrial wastes for the production of amylase and xylanase. Strain MSC702 was identified as novel species of Streptomyces through morphological characterization and 16S rRNA gene sequence. Enzyme production was determined using 1% (w/v) of various agro-industrial waste in production medium containing (g/100mL): K2HPO4(0.1), (NH4)2SO4(0.1), NaCl (0.1), MgSO4(0.1) at pH 7.0 after incubation of 48 h at 50°C. The amylase activity (373.89 IU/mL) and xylanase activity (30.15 IU/mL) was maximum in rice bran. The decreasing order of amylase and xylanase activity in different type of agro-industrial wastes were found rice bran (RB) > corn cob (CC) > wheat bran (WB) > wheat straw (WS) > sugarcane bagasse (SB) and rice bran (RB) > wheat bran (WB) > wheat straw (WS) > sugarcane bagasse (SB) > corn cob (CC), respectively. Mixed effect of different agro-industrial wastes was examined in different ratios. Enzyme yield of amylase and xylanase was ~1.3 and ~2.0 fold higher with RB: WB in 1:2 ratio.