Cellulose hydrolysis by fungi. 1. Screening of cellulolytic strains

Energy Technology Data Exchange (ETDEWEB)

Roussos, S.; Raimbault, M. (Laboratoire de Microbiologie ORSTOM, Centre de Recherche IRCHA, 91 - Vert-le-Petit (France))

Trichoderma harzianum was selected from 30 strains of cellulolytic fungi with the aim of producing cellulases by solid state fermentation of lignocellulosic substrates. Special attention was paid to cellulase production (i. e. carboxymethylcellulase and filter paper activity), apical growth and conidia production. Under the conditions of our experiments, T. harzianum exhibited the highest cellulasic activities with 1,315 IU/I of carboxymethyl cellulose and 80 IU/l of filter paper activity. Apical growth (1 mm/h) and yield of conidial production (3.25 X 10/sup 10/ conidia/g of substrate dry weight) were also valuable characteristics of this strain in the use of solid state fermentation.

Characterization of cellulolytic enzymes and bioH2 production from anaerobic thermophilic Clostridium sp. TCW1.

Science.gov (United States)

Lo, Yung-Chung; Huang, Chi-Yu; Cheng, Chieh-Lun; Lin, Chiu-Yue; Chang, Jo-Shu

2011-09-01

A thermophilic anaerobic bacterium Clostridium sp. TCW1 was isolated from dairy cow dung and was used to produce hydrogen from cellulosic feedstock. Extracellular cellulolytic enzymes produced from TCW1 strain were identified as endoglucanases (45, 53 and 70 kDa), exoglucanase (70 kDa), xylanases (53 and 60 kDa), and β-glucosidase (45 kDa). The endoglucanase and xylanase were more abundant. The optimal conditions for H2 production and enzyme production of the TCW1 strain were the same (60 °C, initial pH 7, agitation rate of 200 rpm). Ten cellulosic feedstock, including pure or natural cellulosic materials, were used as feedstock for hydrogen production by Clostridium strain TCW1 under optimal culture conditions. Using filter paper at 5.0 g/L resulted in the most effective hydrogen production performance, achieving a H2 production rate and yield of 57.7 ml/h/L and 2.03 mol H2/mol hexose, respectively. Production of cellulolytic enzyme activities was positively correlated with the efficiency of dark-H2 fermentation. Copyright © 2011 Elsevier Ltd. All rights reserved.

Thermophillic and thermotolerant fungi isolated from the thermal effluent of nuclear power generating reactors

International Nuclear Information System (INIS)

Rippon, J.W.; Gerhold, R.; Heath, M.

1980-01-01

Over a period of a year, samples of water, foam, microbial mat, soil and air were obtained from areas associated with the cooling canal of a nuclear power station. The seventeen sample sites included water in the cooling canal that was thermally enriched and soil and water adjacent to, up-stream, downstream and at a distance from the generator. Air samples were taken at the plant and at various disstances from the plant. Fifty-two species of thermotolerant and thermophilic fungi were isolated. Of these, eleven species are grouped as opportunistic Mucorales or opportunistic Aspergillus sp. One veterinary pathogen was also isolated (Dactylaria gallopara). The opportunistic/pathogenic fungi were found primarily in the intake bay, the discharge bay and the cooling canal. Smaller numbers were obtained at both upstream and downstream locations. Soil samples near the cooling canal reflected an enrichment of thermophilous organisms, the previously mentioned opportunistic Mucorales and Aspergillus spp. Their numbers were found to be greater than that usually encountered in a mesophilic environment. However, air and soil samples taken at various distances from the power station indicated no greater abundance of these thermophilous fungi than would be expected from a thermal enriched environment. Our results indicate that there was no significant dissemination of thermophilous fungi from the thermal enriched effluents to the adjacent environment. These findings are consistent with the results of other investigators. (orig.)

Thermophilic, lignocellulolytic bacteria for ethanol production: current state and perspectives

DEFF Research Database (Denmark)

Chang, Tinghong; Yao, Shuo

2011-01-01

of cellulolytic and saccharolytic thermophilic bacteria for lignocellulosic ethanol production because of their unique properties. First of all, thermophilic bacteria possess unique cellulolytic and hemicellulolytic systems and are considered as potential sources of highly active and thermostable enzymes...... for efficient biomass hydrolysis. Secondly, thermophilic bacteria ferment a broad range of carbohydrates into ethanol, and some of them display potential for ethanologenic fermentation at high yield. Thirdly, the establishment of the genetic tools for thermophilic bacteria has allowed metabolic engineering......, in particular with emphasis on improving ethanol yield, and this facilitates their employment for ethanol production. Finally, different processes for second-generation ethanol production based on thermophilic bacteria have been proposed with the aim to achieve cost-competitive processes. However, thermophilic...

Effect of two fungicides, benlate and phenyl mercury acetate, on a population of cellulolytic fungi in soil and in pure culture

Energy Technology Data Exchange (ETDEWEB)

Smith, R.N.; Long, P.A.

1980-01-01

Cellulolytic fungi were successfully isolated from an arable loam type soil using the polythene rod technique and the screened substrate technique. Phenyl mercury acetate (PMA) applied to the soil at 100 ppm severely reduced the incidence of many cellulolytic fungi normally present, with a concomitant increase in the incidence of Penicillium spp and Trichoderma lignorum. The application of benlate at 100 ppm had little effect other than to increase the incidence of Doratomyces mircosporus, Dreschlera sp. and Papulospora sp. Both benlate and PMA were much more toxic when examined in-vitro in cellulose agar than when added to soil. Nevertheless, the incidence of cellulolytic isolates from soil reflected the relative sensitivity of the isolates to the respective fungicides in-vitro. The activity of benlate in-vitro decreased with an increase in glucose concentration in the media, but the activity of PMA was independent of glucose concentration.

Thermophilic Fungi to Dominate Aflatoxigenic/Mycotoxigenic Fungi on Food under Global Warming.

Science.gov (United States)

Paterson, Robert Russell M; Lima, Nelson

2017-02-17

Certain filamentous fungi produce mycotoxins that contaminate food. Mycotoxin contamination of crops is highly influenced by environmental conditions and is already affected by global warming, where there is a succession of mycotoxigenic fungi towards those that have higher optimal growth temperatures. Aflatoxigenic fungi are at the highest limit of temperature although predicted increases in temperature are beyond that constraint. The present paper discusses what will succeed these fungi and represents the first such consideration. Aflatoxins are the most important mycotoxins and are common in tropical produce, much of which is exported to temperate regions. Hot countries may produce safer food under climate change because aflatoxigenic fungi will be inhibited. The same situation will occur in previously temperate regions where these fungi have recently appeared, although decades later. Existing thermotolerant and thermophilic fungi (TTF) will dominate, in contrast to the conventional mycotoxigenic fungi adapting or mutating, as it will be quicker. TTF produce a range of secondary metabolites, or potential mycotoxins and patulin which may become a new threat. In addition, Aspergillus fumigatus will appear more frequently, a serious human pathogen, because it is (a) thermotolerant and (b) present on crops: hence this is an even greater problem. An incubation temperature of 41 °C needs employing forthwith to detect TTF. Finally, TTF in crops requires study because of the potential for diseases in humans and animals under climate change.

Diversity of bacteria and glycosyl hydrolase family 48 genes in cellulolytic consortia enriched from thermophilic biocompost.

Science.gov (United States)

Izquierdo, Javier A; Sizova, Maria V; Lynd, Lee R

2010-06-01

The enrichment from nature of novel microbial communities with high cellulolytic activity is useful in the identification of novel organisms and novel functions that enhance the fundamental understanding of microbial cellulose degradation. In this work we identify predominant organisms in three cellulolytic enrichment cultures with thermophilic compost as an inoculum. Community structure based on 16S rRNA gene clone libraries featured extensive representation of clostridia from cluster III, with minor representation of clostridial clusters I and XIV and a novel Lutispora species cluster. Our studies reveal different levels of 16S rRNA gene diversity, ranging from 3 to 18 operational taxonomic units (OTUs), as well as variability in community membership across the three enrichment cultures. By comparison, glycosyl hydrolase family 48 (GHF48) diversity analyses revealed a narrower breadth of novel clostridial genes associated with cultured and uncultured cellulose degraders. The novel GHF48 genes identified in this study were related to the novel clostridia Clostridium straminisolvens and Clostridium clariflavum, with one cluster sharing as little as 73% sequence similarity with the closest known relative. In all, 14 new GHF48 gene sequences were added to the known diversity of 35 genes from cultured species.

Complete genome of the cellulolytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evolutionary adaptations

Energy Technology Data Exchange (ETDEWEB)

Xie, Gary [Los Alamos National Laboratory; Detter, Chris [Los Alamos National Laboratory; Bruce, David [Los Alamos National Laboratory; Challacome, Jean F [Los Alamos National Laboratory; Brettin, Thomas S [Los Alamos National Laboratory; Barabote, Ravi D [UC DAVIS; Leu, David [UC DAVIS; Normand, Philippe [CNRS, UNIV LYON; Necsula, Anamaria [CNRS, UNIV LYON; Daubin, Vincent [CNRS, UNIV LYON; Medigue, Claudine [CNRS/GENOSCOPE; Adney, William S [NREL; Xu, Xin C [UC DAVIS; Lapidus, Alla [DOE JOINT GENOME INST.; Pujic, Pierre [CNRS, UNIV LYON; Richardson, Paul [DOE JOINT GENOME INST; Berry, Alison M [UC DAVIS

2008-01-01

We present here the complete 2.4 MB genome of the actinobacterial thermophile, Acidothermus cellulolyticus lIB, that surprisingly reveals thermophilic amino acid usage in only the cytosolic subproteome rather than its whole proteome. Thermophilic amino acid usage in the partial proteome implies a recent, ongoing evolution of the A. cellulolyticus genome since its divergence about 200-250 million years ago from its closest phylogenetic neighbor Frankia, a mesophilic plant symbiont. Differential amino acid usage in the predicted subproteomes of A. cellulolyticus likely reflects a stepwise evolutionary process of modern thermophiles in general. An unusual occurrence of higher G+C in the non-coding DNA than in the transcribed genome reinforces a late evolution from a higher G+C common ancestor. Comparative analyses of the A. cellulolyticus genome with those of Frankia and other closely-related actinobacteria revealed that A. cellulolyticus genes exhibit reciprocal purine preferences at the first and third codon positions, perhaps reflecting a subtle preference for the dinucleotide AG in its mRNAs, a possible adaptation to a thermophilic environment. Other interesting features in the genome of this cellulolytic, hot-springs dwelling prokaryote reveal streamlining for adaptation to its specialized ecological niche. These include a low occurrence of pseudogenes or mobile genetic elements, a flagellar gene complement previously unknown in this organism, and presence of laterally-acquired genomic islands of likely ecophysiological value. New glycoside hydrolases relevant for lignocellulosic biomass deconstruction were identified in the genome, indicating a diverse biomass-degrading enzyme repertoire several-fold greater than previously characterized, and significantly elevating the industrial value of this organism.

Complete genome of the cellulolytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evolutionary adaptations

Energy Technology Data Exchange (ETDEWEB)

Xie, Gary [Los Alamos National Laboratory; Detter, John C [Los Alamos National Laboratory; Bruce, David C [Los Alamos National Laboratory; Challacombe, Jean F [Los Alamos National Laboratory; Brettin, Thomas S [Los Alamos National Laboratory; Necsulea, Anamaria [UNIV LYON; Daubin, Vincent [UNIV LYON; Medigue, Claudine [GENOSCOPE; Adney, William S [NREL; Xu, Xin C [UC DAVIS; Lapidus, Alla [JGI; Pujic, Pierre [UNIV LYON; Berry, Alison M [UC DAVIS; Barabote, Ravi D [UC DAVIS; Leu, David [UC DAVIS; Normand, Phillipe [UNIV LYON

2009-01-01

We present here the complete 2.4 MB genome of the actinobacterial thermophile, Acidothermus cellulolyticus 11B, that surprisingly reveals thermophilic amino acid usage in only the cytosolic subproteome rather than its whole proteome. Thermophilic amino acid usage in the partial proteome implies a recent, ongoing evolution of the A. cellulolyticus genome since its divergence about 200-250 million years ago from its closest phylogenetic neighbor Frankia, a mesophilic plant symbiont. Differential amino acid usage in the predicted subproteomes of A. cellulolyticus likely reflects a stepwise evolutionary process of modern thermophiles in general. An unusual occurrence of higher G+C in the non-coding DNA than in the transcribed genome reinforces a late evolution from a higher G+C common ancestor. Comparative analyses of the A. cellulolyticus genome with those of Frankia and other closely-related actinobacteria revealed that A. cellulolyticus genes exhibit reciprocal purine preferences at the first and third codon positions, perhaps reflecting a subtle preference for the dinucleotide AG in its mRNAs, a possible adaptation to a thermophilic environment. Other interesting features in the genome of this cellulolytic, hot-springs dwelling prokaryote reveal streamlining for adaptation to its specialized ecological niche. These include a low occurrence of pseudo genes or mobile genetic elements, a flagellar gene complement previously unknown in this organism, and presence of laterally-acquired genomic islands of likely ecophysiological value. New glycoside hydrolases relevant for lignocellulosic biomass deconstruction were identified in the genome, indicating a diverse biomass-degrading enzyme repertoire several-fold greater than previously characterized, and significantly elevating the industrial value of this organism.

Thermophilic Fungi to Dominate Aflatoxigenic/Mycotoxigenic Fungi on Food under Global Warming

Directory of Open Access Journals (Sweden)

Robert Russell M. Paterson

2017-02-01

Full Text Available Certain filamentous fungi produce mycotoxins that contaminate food. Mycotoxin contamination of crops is highly influenced by environmental conditions and is already affected by global warming, where there is a succession of mycotoxigenic fungi towards those that have higher optimal growth temperatures. Aflatoxigenic fungi are at the highest limit of temperature although predicted increases in temperature are beyond that constraint. The present paper discusses what will succeed these fungi and represents the first such consideration. Aflatoxins are the most important mycotoxins and are common in tropical produce, much of which is exported to temperate regions. Hot countries may produce safer food under climate change because aflatoxigenic fungi will be inhibited. The same situation will occur in previously temperate regions where these fungi have recently appeared, although decades later. Existing thermotolerant and thermophilic fungi (TTF will dominate, in contrast to the conventional mycotoxigenic fungi adapting or mutating, as it will be quicker. TTF produce a range of secondary metabolites, or potential mycotoxins and patulin which may become a new threat. In addition, Aspergillus fumigatus will appear more frequently, a serious human pathogen, because it is (a thermotolerant and (b present on crops: hence this is an even greater problem. An incubation temperature of 41 °C needs employing forthwith to detect TTF. Finally, TTF in crops requires study because of the potential for diseases in humans and animals under climate change.

Protease Production by Different Thermophilic Fungi

Science.gov (United States)

Macchione, Mariana M.; Merheb, Carolina W.; Gomes, Eleni; da Silva, Roberto

A comparative study was carried out to evaluate protease production in solid-state fermentation (SSF) and submerged fermentation (SmF) by nine different thermophilic fungi — Thermoascus aurantiacus Miehe, Thermomyces lanuginosus, T. lanuginosus TO.03, Aspergillus flavus 1.2, Aspergillus sp. 13.33, Aspergillus sp. 13.34, Aspergillus sp. 13.35, Rhizomucor pusillus 13.36 and Rhizomucor sp. 13.37 — using substrates containing proteins to induce enzyme secretion. Soybean extract (soybean milk), soybean flour, milk powder, rice, and wheat bran were tested. The most satisfactory results were obtained when using wheat bran in SSF. The fungi that stood out in SSF were T. lanuginosus, T. lanuginosus TO.03, Aspergillus sp. 13.34, Aspergillus sp. 13.35, and Rhizomucor sp. 13.37, and those in SmF were T. aurantiacus, T. lanuginosus TO.03, and 13.37. In both fermentation systems, A. flavus 1.2 and R. pusillus 13.36 presented the lowest levels of proteolytic activity.

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

Science.gov (United States)

Benassi, Vivian Machado; de Lucas, Rosymar Coutinho; Jorge, João Atílio; Polizeli, Maria de Lourdes Teixeira de Moraes

2014-01-01

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 °C, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 °C. 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 °C), while the correspondent extracellular activities were favorably secreted from cultures at 30 °C. 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.

Single-step ethanol production from lignocellulose using novel extremely thermophilic bacteria.

Science.gov (United States)

Svetlitchnyi, Vitali A; Kensch, Oliver; Falkenhan, Doris A; Korseska, Svenja G; Lippert, Nadine; Prinz, Melanie; Sassi, Jamaleddine; Schickor, Anke; Curvers, Simon

2013-02-28

Consolidated bioprocessing (CBP) of lignocellulosic biomass to ethanol using thermophilic bacteria provides a promising solution for efficient lignocellulose conversion without the need for additional cellulolytic enzymes. Most studies on the thermophilic CBP concentrate on co-cultivation of the thermophilic cellulolytic bacterium Clostridium thermocellum with non-cellulolytic thermophilic anaerobes at temperatures of 55°C-60°C. We have specifically screened for cellulolytic bacteria growing at temperatures >70°C to enable direct conversion of lignocellulosic materials into ethanol. Seven new strains of extremely thermophilic anaerobic cellulolytic bacteria of the genus Caldicellulosiruptor and eight new strains of extremely thermophilic xylanolytic/saccharolytic bacteria of the genus Thermoanaerobacter isolated from environmental samples exhibited fast growth at 72°C, extensive lignocellulose degradation and high yield ethanol production on cellulose and pretreated lignocellulosic biomass. Monocultures of Caldicellulosiruptor strains degraded up to 89-97% of the cellulose and hemicellulose polymers in pretreated biomass and produced up to 72 mM ethanol on cellulose without addition of exogenous enzymes. In dual co-cultures of Caldicellulosiruptor strains with Thermoanaerobacter strains the ethanol concentrations rose 2- to 8.2-fold compared to cellulolytic monocultures. A co-culture of Caldicellulosiruptor DIB 087C and Thermoanaerobacter DIB 097X was particularly effective in the conversion of cellulose to ethanol, ethanol comprising 34.8 mol% of the total organic products. In contrast, a co-culture of Caldicellulosiruptor saccharolyticus DSM 8903 and Thermoanaerobacter mathranii subsp. mathranii DSM 11426 produced only low amounts of ethanol. The newly discovered Caldicellulosiruptor sp. strain DIB 004C was capable of producing unexpectedly large amounts of ethanol from lignocellulose in fermentors. The established co-cultures of new Caldicellulosiruptor

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

Directory of Open Access Journals (Sweden)

Vivian Machado Benassi

2014-12-01

Full Text Available 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.

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

Science.gov (United States)

Benassi, Vivian Machado; de Lucas, Rosymar Coutinho; Jorge, João Atílio; Polizeli, Maria de Lourdes Teixeira de Moraes

2014-01-01

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 °C, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 °C. 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 °C), while the correspondent extracellular activities were favorably secreted from cultures at 30 °C. 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. PMID:25763055

Bioprospecting of Thermostable Cellulolytic Enzymes through Modeling and Virtual Screening Method

Directory of Open Access Journals (Sweden)

R. Navanietha Krishnaraj

2017-04-01

Full Text Available Cellulolytic enzymes are promising candidates for the use of cellulose in any bioprocess operations and for the disposal of the cellulosic wastes in an environmentally benign manner. Cellulases from thermophiles have the advantage of hydrolyzing cellulose at wider range of operating conditions unlike the normal enzymes. Herein we report the modeled structures of cellulolytic enzymes (endoglucanase, cellobiohydrolase and ß-glucosidase from a thermophilic bacterium,Clostridium thermocellumand their validation using Root Mean Square Deviation (RMSD and Ramachandran plot analyses. Further, the molecular interactions of the modeled enzyme with cellulose were analyzed using molecular docking technique. The results of molecular docking showed that the endoglucanase, cellobiohydrolase and ß-glucosidase had the binding affinities of -10.7, -9.0 and -10.8 kcal/mol, respectively. A correlation between the binding affinity of the endoglucanase with cellulose and the enzyme activity was also demonstrated. The results showed that the binding affinities of cellulases with cellulose could be used as a tool to assess the hydrolytic activity of cellulases. The results obtained could be used in virtual screening of cellulolytic enzymes based on the molecular interactions with the substrate, and aid in developing systems biology models of thermophiles for industrial biotechnology applications.

Cellulolytic and xylanolytic enzymes from thermophilic Aspergillus terreus RWY.

Science.gov (United States)

Sharma, Reetika; Kocher, Gurvinder Singh; Bhogal, Ravinder Singh; Oberoi, Harinder Singh

2014-12-01

Thermophilic Aspergillus terreus RWY produced cellulases and xylanases in optimal concentrations at 45 °C in solid state fermentation process, though enzyme production was also observed at 50 and 55 °C. Filter paper cellulase (FP), endoglucanase (EG), β-glucosidase (BGL), cellobiohydrolase (CBH), xylanase, β-xylosidase, α-L-arabinofuranosidase and xylan esterase activities for A. terreus RWY at 45 °C in 72 h were 11.3 ± 0.65, 103 ± 6.4, 122.5 ± 8.7, 10.3 ± 0.66, 872 ± 22.5, 22.1 ± 0.75, 126.4 ± 8.4 and 907 ± 15.5 U (g-ds)(-1) , respectively. Enzyme was optimally active at temperatures and pH ranging between 50-60 °C and 4.0-6.0, respectively. The half life (T1/2 ) of 270 and 240 min at 70 and 75 °C, respectively for the enzyme indicates its stability at higher temperatures. The addition of MnCl2 , CoCl2 , and FeCl3 significantly enhanced cellulase activity. Enzyme demonstrated multiplicity by having seven, one and three isoform(s) for EG, CBH and BGL, respectively. Significant production of functionally active consortium of cellulolytic and xylanolytic enzymes from A. terreus RWY makes it a potential candidate in bioprocessing applications. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Screening genus Penicillium for producers of cellulolytic and xylanolytic enzymes

DEFF Research Database (Denmark)

Krogh, Kristian Bertel Rømer; Mørkeberg, Astrid; Frisvad, Jens Christian

2004-01-01

For enzymatic hydrolysis of lignocellulosic material, cellulolytic enzymes from Trichoderma reesei are most commenly used, but, there is a need for more efficient enzyme cocktails. In this study, the production of cellulolytic and xylanolytic enzymes was investigated in 12 filamentous fungi from ...

Efficient plant biomass degradation by thermophilic fungus Myceliophthora heterothallica.

Science.gov (United States)

van den Brink, Joost; van Muiswinkel, Gonny C J; Theelen, Bart; Hinz, Sandra W A; de Vries, Ronald P

2013-02-01

Rapid and efficient enzymatic degradation of plant biomass into fermentable sugars is a major challenge for the sustainable production of biochemicals and biofuels. Enzymes that are more thermostable (up to 70°C) use shorter reaction times for the complete saccharification of plant polysaccharides compared to hydrolytic enzymes of mesophilic fungi such as Trichoderma and Aspergillus species. The genus Myceliophthora contains four thermophilic fungi producing industrially relevant thermostable enzymes. Within this genus, isolates belonging to M. heterothallica were recently separated from the well-described species M. thermophila. We evaluate here the potential of M. heterothallica isolates to produce efficient enzyme mixtures for biomass degradation. Compared to the other thermophilic Myceliophthora species, isolates belonging to M. heterothallica and M. thermophila grew faster on pretreated spruce, wheat straw, and giant reed. According to their protein profiles and in vitro assays after growth on wheat straw, (hemi-)cellulolytic activities differed strongly between M. thermophila and M. heterothallica isolates. Compared to M. thermophila, M. heterothallica isolates were better in releasing sugars from mildly pretreated wheat straw (with 5% HCl) with a high content of xylan. The high levels of residual xylobiose revealed that enzyme mixtures of Myceliophthora species lack sufficient β-xylosidase activity. Sexual crossing of two M. heterothallica showed that progenies had a large genetic and physiological diversity. In the future, this will allow further improvement of the plant biomass-degrading enzyme mixtures of M. heterothallica.

Evaluation of Myceliopthora thermophila as an Enzyme Factory for the Production of Thermophilic Cellulolytic Enzymes

Directory of Open Access Journals (Sweden)

Leonidas Matsakas

2015-07-01

Full Text Available Enzymatic hydrolysis is a key step in bioethanol production. Efficient hydrolysis requires a consortium of different enzymes that are able to hydrolyze cellulose and hemicellulose into fermentable sugars. Myceliopthora thermophila is a promising candidate for the production of thermophilic cellulolytic enzymes, the use of which could reduce the cost of ethanol production. The growth conditions of the fungus were optimized in order to achieve increased secretion of extracellular cellulases. Optimal conditions were found to be 7.0% w/v brewer’s spent grain as the carbon source and 0.4% w/v ammonium sulfate as the nitrogen source. The cellulases obtained were characterized for their optimum activity. The optimum temperature and pH for cellulase activity are 65 °C and pH 5.5, respectively. Studies on thermal inactivation of the crude extract showed that the cellulases of M. thermophila are stable for temperatures up to 60 °C. At this temperature the half-life was found to be as high as 27 h. Enzymatic hydrolysis of cellulose resulted in 31.4% hydrolysis yield at 60 °C after 24 h of incubation. Finally, the recalcitrance constant for cellulose and cellulose pretreated with ionic liquids was calculated to be 5.46 and 2.69, respectively.

Cellulolytic properties of an extremely thermophilic anaerobe

Energy Technology Data Exchange (ETDEWEB)

Hudson, J A; Morgan, H W; Daniel, R M [Waikato Univ., Hamilton (New Zealand). Microbial Biochemistry and Biotechnology Unit

1990-09-01

An extremely thermophilic anaerobe was isolated from a New Zealand hot spring by incubating bacterial mat strands in a medium containing xylan. The Gramreaction-negative organism that was subsequently purified had a temperature optimum of 70deg C and a pH optimum of 7.0. The isolate, designated strain H173, grew on a restricted range of carbon sources. In batch culture H173 could degrade Avicel completely when supplied at 5 or 10 g l{sup -1}. There was an initial growth phase, during which a cellulase complex was produced and carbohydrates fermented to form acetic and lactic acids, followed by a phase where cells were not metabolising but the cellulase complex actively converted cellulose to glucose. When co-cultered with strain Rt8.B1, an ethanologenic extreme thermophile, glucose was fermented to ethanol and acetate, and no reducing sugars accumulated in the medium. In pH controlled batch culture H173 produced an increased amount of lactate and acetate but there was again a phase when reducing sugars accumulated in the medium, and these were converted to ethanol by co-culture with Rt8.B1. (orig.).

Novel glycoside hydrolases from thermophilic fungi

DEFF Research Database (Denmark)

2012-01-01

The present invention relates to isolated polypeptides having cellulolytic activity or hemicellulolytic activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of produ...

Isolation and characterization of a cellulolytic actinomycete Microbispora bispora

Energy Technology Data Exchange (ETDEWEB)

Waldron, Jr, C R; Becker-Vallone, C A; Eveleigh, D E

1986-09-01

Protocols for the isolation of cellulolytic actinomycetes are described, and their use illustrated in the selection of thermophilic bacteria from soil. One isolate, Microbispora bispora, was selected for further study. It grew readily at 55/sup 0/C, produced an extracellular cellulase in good yield (endoglucanase, 5.9 U/ml) that had a broad pH range (pH 5.5 - 7.2) and was thermally stable. Its aryl-..beta..-glucosidase was cell-associated and was relatively resistant to end-product inhibition.

Assessment of cellulolytic microorganisms in soils of Nevados Park, Colombia.

Science.gov (United States)

Avellaneda-Torres, Lizeth Manuela; Pulido, Claudia Patricia Guevara; Rojas, Esperanza Torres

2014-01-01

A systematized survey was conducted to find soil-borne microbes that degrade cellulose in soils from unique ecosystems, such as the Superpáramo, Páramo, and the High Andean Forest in the Nevados National Natural Park (NNNP), Colombia. These high mountain ecosystems represent extreme environments, such as high levels of solar radiation, low atmospheric pressure, and extreme daily changes in temperature. Cellulolytic activity of the microorganisms was evaluated using qualitative tests, such as growth in selective media followed by staining with congo red and iodine, and quantitative tests to determine the activity of endoglucanase, β-glucosidase, exoglucanase, and total cellulase. Microorganisms were identified using molecular markers, such as the 16S rRNA gene for bacteria and the internal transcribed spacer region (ITS) of ribosomal DNA for fungi. Multivariate statistical analysis (MVA) was used to select microorganisms with high cellulolytic capacity. A total of 108 microorganisms were isolated from the soils and, in general, the enzymatic activities of fungi were higher than those of bacteria. Our results also found that none of the organisms studied were able to degrade all the components of the cellulose and it is therefore suggested that a combination of bacteria and/or fungi with various enzymatic activities be used to obtain high total cellulolytic activity. This study gives an overview of the potential microorganism that could be used for cellulose degradation in various biotechnological applications and for sustainable agricultural waste treatment.

Assessment of cellulolytic microorganisms in soils of Nevados Park, Colombia

Directory of Open Access Journals (Sweden)

Lizeth Manuela Avellaneda-Torres

2014-12-01

Full Text Available A systematized survey was conducted to find soil-borne microbes that degrade cellulose in soils from unique ecosystems, such as the Superpáramo, Páramo, and the High Andean Forest in the Nevados National Natural Park (NNNP, Colombia. These high mountain ecosystems represent extreme environments, such as high levels of solar radiation, low atmospheric pressure, and extreme daily changes in temperature. Cellulolytic activity of the microorganisms was evaluated using qualitative tests, such as growth in selective media followed by staining with congo red and iodine, and quantitative tests to determine the activity of endoglucanase, β-glucosidase, exoglucanase, and total cellulase. Microorganisms were identified using molecular markers, such as the 16S rRNA gene for bacteria and the internal transcribed spacer region (ITS of ribosomal DNA for fungi. Multivariate statistical analysis (MVA was used to select microorganisms with high cellulolytic capacity. A total of 108 microorganisms were isolated from the soils and, in general, the enzymatic activities of fungi were higher than those of bacteria. Our results also found that none of the organisms studied were able to degrade all the components of the cellulose and it is therefore suggested that a combination of bacteria and/or fungi with various enzymatic activities be used to obtain high total cellulolytic activity. This study gives an overview of the potential microorganism that could be used for cellulose degradation in various biotechnological applications and for sustainable agricultural waste treatment.

Cellulolytic and xylanolytic activities of common indoor fungi

DEFF Research Database (Denmark)

Andersen, Birgitte; Poulsen, Rehab; Hansen, Gustav Hammerich

2016-01-01

Moldy building materials, such as chip wood and gypsum, should be a good source for fungal strains with high production of lignocellulolytic enzymes. Screening of 21 common indoor fungal strains showed, contrary to the expected, that the Chaetomium and Stachybotrys strains had little...... or no cellulolytic and xylanolytic activities using AZCL-assays. On the other hand, both Cladosporium sphaerospermum and Penicillium chrysogenum showed the highest cellulase, β-glucosidase, mannase, β-galactanase and arabinanase activities and would be good candidates for over-producers of enzymes needed...

Characterization of thermophilic fungal community associated with pile fermentation of Pu-erh tea.

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Zhang, Wei; Yang, Ruijuan; Fang, Wenjun; Yan, Liang; Lu, Jun; Sheng, Jun; Lv, Jie

2016-06-16

This study aimed to characterize the thermophilic fungi in pile-fermentation process of Pu-erh tea. Physicochemical analyses showed that the high temperature and low pH provided optimal conditions for propagation of fungi. A number of fungi, including Blastobotrys adeninivorans, Thermomyces lanuginosus, Rasamsonia emersonii, Aspergillus fumigatus, Rhizomucor pusillus, Rasamsonia byssochlamydoides, Rasamsonia cylindrospora, Aspergillus tubingensis, Aspergillus niger, Candida tropicalis and Fusarium graminearum were isolated as thermophilic fungi under combination of high temperature and acid culture conditions from Pu-erh tea pile-fermentation. The fungal communities were analyzed by PCR-DGGE. Results revealed that those fungi are closely related to Debaryomyces hansenii, Cladosporium cladosporioides, A. tubingensis, R. emersonii, R. pusillus, A. fumigatus and A. niger, and the last four presented as dominant species in the pile process. These four preponderant thermophilic fungi reached the order of magnitude of 10(7), 10(7), 10(7) and 10(6)copies/g dry tea, respectively, measured by real-time quantitative PCR (q-PCR). The results indicate that the thermophilic fungi play an important role in Pu-erh tea pile fermentation. Copyright © 2016 Elsevier B.V. All rights reserved.

Cellulolytic activities of wild type fungi isolated from decayed wood ...

African Journals Online (AJOL)

Prof. Ogunji

amongst the fungal isolates while M. mucedo had the least cellulolytic ... across plant taxa, high cellulose content; typically in the range of ... antibiotic mixture made up of distilled water, 50 ml and Erythromycin: 500mg was .... cellulases including β-glucosidase were produced from Penicillium, Aspergillus and Trichoderma ...

Cellulosic ethanol production via consolidated bioprocessing by a novel thermophilic anaerobic bacterium isolated from a Himalayan hot spring.

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Singh, Nisha; Mathur, Anshu S; Tuli, Deepak K; Gupta, Ravi P; Barrow, Colin J; Puri, Munish

2017-01-01

Cellulose-degrading thermophilic anaerobic bacterium as a suitable host for consolidated bioprocessing (CBP) has been proposed as an economically suited platform for the production of second-generation biofuels. To recognize the overall objective of CBP, fermentation using co-culture of different cellulolytic and sugar-fermenting thermophilic anaerobic bacteria has been widely studied as an approach to achieving improved ethanol production. We assessed monoculture and co-culture fermentation of novel thermophilic anaerobic bacterium for ethanol production from real substrates under controlled conditions. In this study, Clostridium sp. DBT-IOC-C19, a cellulose-degrading thermophilic anaerobic bacterium, was isolated from the cellulolytic enrichment cultures obtained from a Himalayan hot spring. Strain DBT-IOC-C19 exhibited a broad substrate spectrum and presented single-step conversion of various cellulosic and hemicellulosic substrates to ethanol, acetate, and lactate with ethanol being the major fermentation product. Additionally, the effect of varying cellulose concentrations on the fermentation performance of the strain was studied, indicating a maximum cellulose utilization ability of 10 g L -1 cellulose. Avicel degradation kinetics of the strain DBT-IOC-C19 displayed 94.6% degradation at 5 g L -1 and 82.74% degradation at 10 g L -1 avicel concentration within 96 h of fermentation. In a comparative study with Clostridium thermocellum DSM 1313, the ethanol and total product concentrations were higher by the newly isolated strain on pretreated rice straw at an equivalent substrate loading. Three different co-culture combinations were used on various substrates that presented two-fold yield improvement than the monoculture during batch fermentation. This study demonstrated the direct fermentation ability of the novel thermophilic anaerobic bacteria on various cellulosic and hemicellulosic substrates into ethanol without the aid of any exogenous enzymes

Effect of cellulase producing fungi on plant residues degradation used as organic fertilizer

International Nuclear Information System (INIS)

Ibrahim, R.M.M

2009-01-01

Series of laboratory and field experiments were conducted at Soil microbiology Unit and Farm of soil and Water research department, Nuclear Research Center, Atomic Energy Authority, Egypt. Laboratory experiments revealed that between nine fungal strain, A. niger was the most potent cellulolytic fungus able to degrade many cellulosic sources (CP, CMC, and FP). Study the effect of cellulolytic fungi on degradation of plant residues used as organic fertilizer in addition to nitrogen fixing bacteria (symbiotically) on lupine growth, yield and nutrients uptake (Field experiment) had been carried out. This objective aims to recycling different plant residues in soil which is consistent with (sustainable development) and utilization of these organic residues as a single carbon source for cellulolytic fungi.Application of 15 N- tracer technique gave us the chance and opportunity to quantify the exact amounts of N derived from the different sources of nitrogen available to lupine plant under the effect of cellulolytic fungi on different plant residues.The obtained results could be summarized as following:I.Laboratory Technique Selection of the most potent cellulolytic fungi 1-Nine fungal strains of Aspergillus niger; Penicillium oxalicum; Trichoderma longibranchiatum; Aspergillus terreus; Aspergillus flavus; Alterrnaria sp.; Trichderma harzianum ; Rhizopus sp. and Syncephalastrum sp. obtained from different sources and tested for their cellulolytic activity. 2-Aspergillus niger and Pencillium oxalicum exhibited the highest cellulase productivity followed by Trichoderma longibranchiatum and Aspergillus terreus.3- fungal mixtures of the most potent four genera Aspergillus niger; Penicillium oxalicum; Trichoderma longibranchiatum and Aspergillus terreus found to have a lower cellulolytic activities for all substrates compared with single inoculation with A. niger.4-Highest FPase activities were exhibited by A. niger when filter paper (FP) used as a carbon source.5-A. niger is

Mixed submerged fermentation with two filamentous fungi for cellulolytic and xylanolytic enzyme production.

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Garcia-Kirchner, O; Muñoz-Aguilar, M; Pérez-Villalva, R; Huitrón-Vargas, C

2002-01-01

The efficient saccharification of lignocellulosic materials requires the cooperative actions of different cellulase enzyme activities: exoglucanase, endoglucanase, beta-glucosidase, and xylanase. Previous studies with the fungi strains Aureobasidium sp. CHTE-18, Penicillium sp. CH-TE-001, and Aspergillus terreus CH-TE-013, selected mainly because of their different cellulolytic and xylanolytic activities, have demonstrated the capacity of culture filtrates of cross-synergistic action in the saccharification of native sugarcane bagasse pith. In an attempt to improve the enzymatic hydrolysis of different cellulosic materials, we investigated a coculture fermentation with two of these strains to enhance the production of cellulases and xylanases. The 48-h batch experimental results showed that the mixed culture of Penicillium sp. CH-TE-001 and A. terreus CH-TE-013 produced culture filtrates with high protein content, cellulase (mainly beta-glucosidase), and xylanase activities compared with the individual culture of each strain. The same culture conditions were used in a simple medium with mineral salts, corn syrup liquor, and sugarcane bagasse pith as the sole carbon source with moderate shaking at 29 degrees C. Finally, we compared the effect of the cell-free culture filtrates obtained from the mixed and single fermentations on the saccharification of different kinds of cellulosic materials.

Utilizing thermophilic microbe in lignocelluloses based bioethanol production: Review

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Sriharti, Agustina, Wawan; Ratnawati, Lia; Rahman, Taufik; Salim, Takiyah

2017-01-01

The utilization of thermophilic microbe has attracted many parties, particularly in producing an alternative fuel like ethanol. Bioethanol is one of the alternative energy sources substituting for earth oil in the future. The advantage of using bioethanol is that it can reduce pollution levels and global warming because the result of bioethanol burning doesn't bring in a net addition of CO2 into environment. Moreover, decrease in the reserves of earth oil globally has also contributed to the notion on searching renewable energy resources such as bioethanol. Indonesia has a high biomass potential and can be used as raw material for bioethanol. The utilization of these raw materials will reduce fears of competition foodstuffs for energy production. The enzymes that play a role in degrading lignocelluloses are cellulolytic, hemicellulolytic, and lignolytic in nature. The main enzyme with an important role in bioethanol production is a complex enzyme capable of degrading lignocelluloses. The enzyme can be produced by the thermophilik microbes of the groups of bacteria and fungi such as Trichoderma viride, Clostridium thermocellum, Bacillus sp. Bioethanol production is heavily affected by raw material composition, microorganism type, and the condition of fermentation used.

Caldicellulosiruptor obsidiansis sp. nov., an anaerobic, extremely thermophilic, cellulolytic bacterium isolated from Obsidian Pool, Yellowstone National Park.

Science.gov (United States)

Hamilton-Brehm, Scott D; Mosher, Jennifer J; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J; Keller, Martin; Elkins, James G

2010-02-01

A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47(T), was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 microm long by 0.2 microm wide and grew at temperatures between 55 and 85 degrees C, with the optimum at 78 degrees C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h(-1). The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47(T) was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47(T) within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073).

Caldicellulosiruptor obsidiansis sp. nov., an Anaerobic, Extremely Thermophilic, Cellulolytic Bacterium Isolated from Obsidian Pool, Yellowstone National Park▿

Science.gov (United States)

Hamilton-Brehm, Scott D.; Mosher, Jennifer J.; Vishnivetskaya, Tatiana; Podar, Mircea; Carroll, Sue; Allman, Steve; Phelps, Tommy J.; Keller, Martin; Elkins, James G.

2010-01-01

A novel, obligately anaerobic, extremely thermophilic, cellulolytic bacterium, designated OB47T, was isolated from Obsidian Pool, Yellowstone National Park, WY. The isolate was a nonmotile, non-spore-forming, Gram-positive rod approximately 2 μm long by 0.2 μm wide and grew at temperatures between 55 and 85°C, with the optimum at 78°C. The pH range for growth was 6.0 to 8.0, with values of near 7.0 being optimal. Growth on cellobiose produced the fastest specific growth rate at 0.75 h−1. The organism also displayed fermentative growth on glucose, maltose, arabinose, fructose, starch, lactose, mannose, sucrose, galactose, xylose, arabinogalactan, Avicel, xylan, filter paper, processed cardboard, pectin, dilute acid-pretreated switchgrass, and Populus. OB47T was unable to grow on mannitol, fucose, lignin, Gelrite, acetate, glycerol, ribose, sorbitol, carboxymethylcellulose, and casein. Yeast extract stimulated growth, and thiosulfate, sulfate, nitrate, and sulfur were not reduced. Fermentation end products were mainly acetate, H2, and CO2, although lactate and ethanol were produced in 5-liter batch fermentations. The G+C content of the DNA was 35 mol%, and sequence analysis of the small subunit rRNA gene placed OB47T within the genus Caldicellulosiruptor. Based on its phylogenetic and phenotypic properties, the isolate is proposed to be designated Caldicellulosiruptor obsidiansis sp. nov. and OB47 is the type strain (ATCC BAA-2073). PMID:20023107

Thermophilic growth and enzymatic thermostability are polyphyletic traits within Chaetomiaceae.

Science.gov (United States)

van den Brink, Joost; Facun, Kryss; de Vries, Michel; Stielow, J Benjamin

2015-12-01

Thermophilic fungi have the potential to produce industrial-relevant thermostable enzymes, in particular for the degradation of plant biomass. Sordariales is one of the few fungal orders containing several thermophilic taxa, of which many have been associated with the production of thermostable enzymes. The evolutionary affiliation of Sordariales fungi, especially between thermophiles and non-thermophilic relatives, is however poorly understood. Phylogenetic analysis within the current study was based on sequence data, derived from a traditional Sanger and highly multiplexed targeted next generation sequencing approach of 45 isolates. The inferred phylogeny and detailed growth analysis rendered the trait 'thermophily' as polyphyletic within Chaetomiaceae (Sordariales, Sordariomycetes), and characteristic to: Myceliophthora spp., Thielavia terrestris, Chaetomium thermophilum, and Mycothermus thermophilus. Compared to mesophiles, the isolates within thermophilic taxa produced enzyme mixtures with the highest thermostability of known cellulase activities. Temperature profiles of the enzyme activities correlated strongly with the optimal growth temperatures of the isolates but not with their phylogenetic relationships. This strong correlation between growth and enzyme characteristics indicated that detailed analysis of growth does give predictive information on enzyme physiology. The variation in growth and enzyme characteristics reveals these fungi as an excellent platform to better understand fungal thermophily and enzyme thermostability. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Bacillus coagulans MA-13: a promising thermophilic and cellulolytic strain for the production of lactic acid from lignocellulosic hydrolysate.

Science.gov (United States)

Aulitto, Martina; Fusco, Salvatore; Bartolucci, Simonetta; Franzén, Carl Johan; Contursi, Patrizia

2017-01-01

The transition from a petroleum-based economy towards more sustainable bioprocesses for the production of fuels and chemicals (circular economy) is necessary to alleviate the impact of anthropic activities on the global ecosystem. Lignocellulosic biomass-derived sugars are suitable alternative feedstocks that can be fermented or biochemically converted to value-added products. An example is lactic acid, which is an essential chemical for the production of polylactic acid, a biodegradable bioplastic. However, lactic acid is still mainly produced by Lactobacillus species via fermentation of starch-containing materials, the use of which competes with the supply of food and feed. A thermophilic and cellulolytic lactic acid producer was isolated from bean processing waste and was identified as a new strain of Bacillus coagulans , named MA-13. This bacterium fermented lignocellulose-derived sugars to lactic acid at 55 °C and pH 5.5. Moreover, it was found to be a robust strain able to tolerate high concentrations of hydrolysate obtained from wheat straw pre-treated by acid-catalysed (pre-)hydrolysis and steam explosion, especially when cultivated in controlled bioreactor conditions. Indeed, unlike what was observed in microscale cultivations (complete growth inhibition at hydrolysate concentrations above 50%), B. coagulans MA-13 was able to grow and ferment in 95% hydrolysate-containing bioreactor fermentations. This bacterium was also found to secrete soluble thermophilic cellulases, which could be produced at low temperature (37 °C), still retaining an optimal operational activity at 50 °C. The above-mentioned features make B. coagulans MA-13 an appealing starting point for future development of a consolidated bioprocess for production of lactic acid from lignocellulosic biomass, after further strain development by genetic and evolutionary engineering. Its optimal temperature and pH of growth match with the operational conditions of fungal enzymes hitherto

The significance of cellulolytic enzymes produced by Trichoderma in opportunistic lifestyle of this fungus.

Science.gov (United States)

Strakowska, Judyta; Błaszczyk, Lidia; Chełkowski, Jerzy

2014-07-01

The degradation of native cellulose to glucose monomers is a complex process, which requires the synergistic action of the extracellular enzymes produced by cellulolytic microorganisms. Among fungi, the enzymatic systems that can degrade native cellulose have been extensively studied for species belonging to the genera of Trichoderma. The majority of the cellulolytic enzymes described so far have been examples of Trichoderma reesei, extremely specialized in the efficient degradation of plant cell wall cellulose. Other Trichoderma species, such as T. harzianum, T. koningii, T. longibrachiatum, and T. viride, known for their capacity to produce cellulolytic enzymes, have been isolated from various ecological niches, where they have proved successful in various heterotrophic interactions. As saprotrophs, these species are considered to make a contribution to the degradation of lignocellulosic plant material. Their cellulolytic potential is also used in interactions with plants, especially in plant root colonization. However, the role of cellulolytic enzymes in species forming endophytic associations with plants or in those existing in the substratum for mushroom cultivation remains unknown. The present review discusses the current state of knowledge about cellulolytic enzymes production by Trichoderma species and the encoding genes, as well as the involvement of these proteins in the lifestyle of Trichoderma. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cellulose- and xylan-degrading thermophilic anaerobic bacteria from biocompost.

Science.gov (United States)

Sizova, M V; Izquierdo, J A; Panikov, N S; Lynd, L R

2011-04-01

Nine thermophilic cellulolytic clostridial isolates and four other noncellulolytic bacterial isolates were isolated from self-heated biocompost via preliminary enrichment culture on microcrystalline cellulose. All cellulolytic isolates grew vigorously on cellulose, with the formation of either ethanol and acetate or acetate and formate as principal fermentation products as well as lactate and glycerol as minor products. In addition, two out of nine cellulolytic strains were able to utilize xylan and pretreated wood with roughly the same efficiency as for cellulose. The major products of xylan fermentation were acetate and formate, with minor contributions of lactate and ethanol. Phylogenetic analyses of 16S rRNA and glycosyl hydrolase family 48 (GH48) gene sequences revealed that two xylan-utilizing isolates were related to a Clostridium clariflavum strain and represent a distinct novel branch within the GH48 family. Both isolates possessed high cellulase and xylanase activity induced independently by either cellulose or xylan. Enzymatic activity decayed after growth cessation, with more-rapid disappearance of cellulase activity than of xylanase activity. A mixture of xylan and cellulose was utilized simultaneously, with a significant synergistic effect observed as a reduction of lag phase in cellulose degradation.

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

Science.gov (United States)

Aanniz, Tarik; Ouadghiri, Mouna; Melloul, Marouane; Swings, Jean; Elfahime, Elmostafa; Ibijbijen, Jamal; Ismaili, Mohamed; Amar, Mohamed

2015-06-01

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.

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

Directory of Open Access Journals (Sweden)

Tarik Aanniz

2015-06-01

Full Text Available 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.

Exogenous cellulases of thermophilic micromycetes. Pt. 2. Thermostability of enzyme preparations

Energy Technology Data Exchange (ETDEWEB)

Kvesitadze, G; Gogilashvili, L; Svanidze, R; Buachidze, T; Chirgadze, L; Nizharadze, D

1986-01-01

The ability of a large number of higher fungi to form extracellular cellulases is investigated. Some representatives of these fungi grow at 40-50/sup 0/C, and form extracellular cellulases exceeding cellulases of mesophilic fungi in thermostability. It is shown that cellulases of higher thermophilic fungi differ by their thermostability. The temperature optimum of cellulase action of higher fungi occurs within 60-62/sup 0/C.

Pyrosequencing reveals high-temperature cellulolytic microbial consortia in Great Boiling Spring after in situ lignocellulose enrichment.

Directory of Open Access Journals (Sweden)

Joseph P Peacock

Full Text Available To characterize high-temperature cellulolytic microbial communities, two lignocellulosic substrates, ammonia fiber-explosion-treated corn stover and aspen shavings, were incubated at average temperatures of 77 and 85°C in the sediment and water column of Great Boiling Spring, Nevada. Comparison of 109,941 quality-filtered 16S rRNA gene pyrosequences (pyrotags from eight enrichments to 37,057 quality-filtered pyrotags from corresponding natural samples revealed distinct enriched communities dominated by phylotypes related to cellulolytic and hemicellulolytic Thermotoga and Dictyoglomus, cellulolytic and sugar-fermenting Desulfurococcales, and sugar-fermenting and hydrogenotrophic Archaeoglobales. Minor enriched populations included close relatives of hydrogenotrophic Thermodesulfobacteria, the candidate bacterial phylum OP9, and candidate archaeal groups C2 and DHVE3. Enrichment temperature was the major factor influencing community composition, with a negative correlation between temperature and richness, followed by lignocellulosic substrate composition. This study establishes the importance of these groups in the natural degradation of lignocellulose at high temperatures and suggests that a substantial portion of the diversity of thermophiles contributing to consortial cellulolysis may be contained within lineages that have representatives in pure culture.

Kallotenue papyrolyticum gen. nov., sp. nov., a cellulolytic and filamentous thermophile that represents a novel lineage (Kallotenuales ord. nov., Kallotenuaceae fam. nov.) within the class Chloroflexia

Energy Technology Data Exchange (ETDEWEB)

Cole, Jesse; Gieler, Brandon; Heisler, Devon; Palisoc, Maryknoll; Williams, Amanda; Dohnalkova, Alice; Ming, Hong; Yu, Tian T.; Dodsworth, Jeremy A.; Li, Wen J.; Hedlund, Brian P.

2013-08-15

Several closely-related, thermophilic, and cellulolytic bacterial strains, designated JKG1T, JKG2, JKG3, JKG4, and JKG5, were isolated from a cellulolytic enrichment (corn stover) incubated in the water column of Great Boiling Spring, NV. Strain JKG1T had cells of a diameter of 0.7 - 0.9 μm and length of ~2.0 μm that formed non-branched multicellular filaments reaching >300 μm. Spores were not formed and dense liquid cultures were red. The temperature range for growth was 45-65 °C, with an optimum of 55 °C. The pH range for growth was 5.6-9.0, with an optimum of 7.5. JKG1T grew as an aerobic heterotroph, utilizing glucose, sucrose, xylose, arabinose, cellobiose, carboxymethylcellulose, filter paper, microcrystalline cellulose, xylan, starch, casamino acids, tryptone, peptone, yeast extract, acetate, citrate, lactate, pyruvate, and glycerol as sole carbon sources, and was not observed to photosynthesize. The cells stained Gram-negative. Phylogenetic analysis using 16S rRNA gene sequences placed the new isolates in the class Chloroflexia, but distant from other cultivated members, with the highest sequence identity of 82.5% to Roseiflexus castenholzii. The major quinone was menaquinone-9; no ubiquinones were detected. The major cellular fatty acids (>5%) were C18:0, anteiso-C17:0, iso-C18:0, and iso-C17:0. C16:0, iso-C16:0, and C17:0. The peptidoglycan amino acids were alanine, ornithine, glutamic acid, serine, and asparagine. Whole-cell sugars included mannose, rhamnose, glucose, galactose, ribose, arabinose, and xylose. Morphological, phylogenetic, and chemotaxonomic results suggest that JKG1T is representative of a new lineage within the class Chloroflexia, which we propose to designate Kallotenue papyrolyticum gen. nov., sp. nov., Kallotenuaceae fam. nov., Kallotenuales ord. nov.

New thermophilic anaerobes that decompose crystalline cellulose

Energy Technology Data Exchange (ETDEWEB)

Taya, M; Hinoki, H; Suzuki, Y; Yagi, T; Yap, M G.S.; Kobayashi, T

1985-01-01

Two strains (designated as 25A and 3B) of cellulolytic, thermophilic, anaerobic, spore-forming bacteria were newly isolated from an alkaline hot spring through enrichment cultures at 60/sup 0/C. Though strain 25A was nearly identical to Clostridium thermocellum ATCC 27405 as a reference strain, strain 3B had some characteristics different from the reference; no flagellation, alkalophilic growth property (optimum pH of 7.5-8) and orange-colored pigmentation of the cell mass. Strain 3B effectively decomposed micro-crystalline cellulose (Avicel) and raw cellulosics (rice straw, newspaper, and bagasse) without physical or chemical pretreatments. 20 references, 2 figures, 2 tables.

Discrete and structurally unique proteins (tāpirins) mediate attachment of extremely thermophilic Caldicellulosiruptor species to cellulose.

Science.gov (United States)

Blumer-Schuette, Sara E; Alahuhta, Markus; Conway, Jonathan M; Lee, Laura L; Zurawski, Jeffrey V; Giannone, Richard J; Hettich, Robert L; Lunin, Vladimir V; Himmel, Michael E; Kelly, Robert M

2015-04-24

A variety of catalytic and noncatalytic protein domains are deployed by select microorganisms to deconstruct lignocellulose. These extracellular proteins are used to attach to, modify, and hydrolyze the complex polysaccharides present in plant cell walls. Cellulolytic enzymes, often containing carbohydrate-binding modules, are key to this process; however, these enzymes are not solely responsible for attachment. Few mechanisms of attachment have been discovered among bacteria that do not form large polypeptide structures, called cellulosomes, to deconstruct biomass. In this study, bioinformatics and proteomics analyses identified unique, discrete, hypothetical proteins ("tāpirins," origin from Māori: to join), not directly associated with cellulases, that mediate attachment to cellulose by species in the noncellulosomal, extremely thermophilic bacterial genus Caldicellulosiruptor. Two tāpirin genes are located directly downstream of a type IV pilus operon in strongly cellulolytic members of the genus, whereas homologs are absent from the weakly cellulolytic Caldicellulosiruptor species. Based on their amino acid sequence, tāpirins are specific to these extreme thermophiles. Tāpirins are also unusual in that they share no detectable protein domain signatures with known polysaccharide-binding proteins. Adsorption isotherm and trans vivo analyses demonstrated the carbohydrate-binding module-like affinity of the tāpirins for cellulose. Crystallization of a cellulose-binding truncation from one tāpirin indicated that these proteins form a long β-helix core with a shielded hydrophobic face. Furthermore, they are structurally unique and define a new class of polysaccharide adhesins. Strongly cellulolytic Caldicellulosiruptor species employ tāpirins to complement substrate-binding proteins from the ATP-binding cassette transporters and multidomain extracellular and S-layer-associated glycoside hydrolases to process the carbohydrate content of lignocellulose. �

Concomitant production of cellulase and xylanase by thermophilic mould Sporotrichum thermophile in solid state fermentation and their applicability in bread making.

Science.gov (United States)

Bala, Anju; Singh, Bijender

2017-06-01

Sporotrichum thermophile BJAMDU5 secreted high titres of xylanolytic and cellulolytic enzymes in solid state fermentation using mixture of wheat straw and cotton oil cake (ratio 1:1) at 45 °C, pH 5.0 after 72 h inoculated with 2.9 × 10 7  CFU/mL conidiospores. Supplementation of solid medium with lactose and ammonium sulphate further enhanced the production of hydrolytic enzymes. Among different surfactants studied, Tween 80 enhanced the production of all enzymes [3455 U/g DMR (dry mouldy residue), 879.26 U/g DMR, 976.28 U/g DMR and 35.10 U/g DMR for xylanase, CMCase (Carboxymethylcellulase), FPase (Filter paper activity) and β-glucosidase, respectively] as compared to other surfactants. Recycling of solid substrate reduced the production of all these enzymes after second cycle. End products analysis by TLC showed the ability of hydrolytic enzymes of S. thermophile to liberate monomeric (xylose and glucose) as well as oligomeric (xylobiose, cellobiose and higher ones) sugars. Supplementation of enzyme resulted in improved nutritional properties of the bread. Formation of oligomeric sugars by xylanase enzyme of S. thermophile BJAMDU5 make it a good candidate in food industry.

Thermophilic fungi as new sources for production of cellulases and xylanases with potential use in sugarcane bagasse saccharification.

Science.gov (United States)

de Cassia Pereira, J; Paganini Marques, N; Rodrigues, A; Brito de Oliveira, T; Boscolo, M; da Silva, R; Gomes, E; Bocchini Martins, D A

2015-04-01

To obtain new cellulases and xylanases from thermophilic fungi; evaluate their potential for sugarcane bagasse saccharification. Thirty-two heat-tolerant fungi were isolated from the environment, identified (morphological/molecular tools) and the production of the enzymes was evaluated by solid state fermentation using lignocellulosic materials as substrates. Myceliophthora thermophila JCP 1-4 was the best producer of endoglucanase (357·51 U g(-1) ), β-glucosidase (45·42 U g(-1) ), xylanase (931·11 U g(-1) ) and avicelase (3·58 U g(-1) ). These enzymes were most active at 55-70°C and stable at 30-60°C. Using crude enzymatic extract from M. thermophila JCP 1-4 to saccharify sugarcane bagasse pretreated with microwaves and glycerol, glucose and xylose yields obtained were 15·6 and 35·13% (2·2 and 1·95 g l(-1) ), respectively. All isolated fungi have potential to produce the enzymes; M. thermophila JCP 1-4 enzymatic extract have potential to be better explored in saccharification experiments. Pretreatment improved enzymatic saccharification, as sugar yields were much higher than those obtained from in natura bagasse. Myceliophthora thermophila JCP 1-4 produces avicelase (not commonly found among fungi; important to hydrolyse crystalline cellulose) and a β-glucosidase resistant to glucose inhibition, interesting characteristics for saccharification experiments. © 2015 The Society for Applied Microbiology.

Cellulolytic potential under environmental changes in microbial communities from grassland litter

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Renaud eBerlemont

2014-11-01

Full Text Available In many ecosystems, global changes are likely to profoundly affect microorganisms. In Southern California, changes in precipitation and nitrogen deposition may influence the composition and functional potential of microbial communities and their resulting ability to degrade plant material. To test whether environmental changes impact the distribution of functional groups involved in leaf litter degradation, we determined how the genomic diversity of microbial communities in a semi-arid grassland ecosystem changed under reduced precipitation or increased N deposition. We monitored communities seasonally over a period of two years to place environmental change responses into the context of natural variation. Fungal and bacterial communities displayed strong seasonal patterns, Fungi being mostly detected during the dry season whereas Bacteria were common during wet periods. Most putative cellulose degraders were associated with 33 bacterial genera and constituted ~18.2% of the microbial community. Precipitation reduction reduced bacterial abundance and cellulolytic potential whereas nitrogen addition did not affect the cellulolytic potential of the microbial community. Finally, we detected a strong correlation between the frequencies of genera putative cellulose degraders and cellulase genes. Thus, microbial taxonomic composition was predictive of cellulolytic potential. This work provides a framework for how environmental changes affect microorganisms responsible for plant litter deconstruction.

Metaproteomics reveals major microbial players and their biodegradation functions in a large-scale aerobic composting plant

Science.gov (United States)

Liu, Dongming; Li, Mingxiao; Xi, Beidou; Zhao, Yue; Wei, Zimin; Song, Caihong; Zhu, Chaowei

2015-01-01

Composting is an appropriate management alternative for municipal solid waste; however, our knowledge about the microbial regulation of this process is still scare. We employed metaproteomics to elucidate the main biodegradation pathways in municipal solid waste composting system across the main phases in a large-scale composting plant. The investigation of microbial succession revealed that Bacillales, Actinobacteria and Saccharomyces increased significantly with respect to abundance in composting process. The key microbiologic population for cellulose degradation in different composting stages was different. Fungi were found to be the main producers of cellulase in earlier phase. However, the cellulolytic fungal communities were gradually replaced by a purely bacterial one in active phase, which did not support the concept that the thermophilic fungi are active through the thermophilic phase. The effective decomposition of cellulose required the synergy between bacteria and fungi in the curing phase. PMID:25989417

Selection and molecular characterization of cellulolytic-xylanolytic fungi from surface soil-biomass mixtures from Black Belt sites.

Science.gov (United States)

Okeke, Benedict C; Hall, Rosine W; Nanjundaswamy, Ananda; Thomson, M Sue; Deravi, Yasaman; Sawyer, Leah; Prescott, Andrew

2015-06-01

Plant biomass is an abundant renewable natural resource that can be transformed into chemical feedstocks. Enzymes used in saccharification of lignocellulosic biomass are a major part of biofuel production costs. A cocktail of cellulolytic and xylanolytic enzymes are required for optimal saccharification of biomass. Accordingly, thirty-two fungal pure cultures were obtained from surface soil-biomass mixtures collected from Black Belt sites in Alabama by culturing on lignocellulosic biomass medium. The fungal strains were screened for the coproduction of cellulolytic and xylanolytic enzymes. Strains that displayed promising levels of cellulolytic and xylanolytic enzymes were characterized by molecular analysis of DNA sequences from the large subunit and internal transcribed spacer (ITS) of their ribosomal RNA gene. Nucleotide sequence analysis revealed that two most promising isolates FS22A and FS5A were most similar to Penicillium janthinellum and Trichoderma virens. Production dynamics of cellulolytic and xylanolytic enzymes from these two strains were explored in submerged fermentation. Volumetric productivity after 120 h incubation was 121.08 units/L/h and 348 units/L/h for the filter paper cellulase and xylanase of strain FS22A, and 90.83 units/L/h and 359 units/L/h, respectively for strain FS5A. Assays with 10 times dilution of enzymes revealed that the activities were much higher than that observed in the crude culture supernatant. Additionally, both FS22A and FS5A also produced amylase in lignocellulose medium. The enzyme profiles of these strains and their activities suggest potential applications in cost effective biomass conversion and biodegradation. Copyright © 2015 Elsevier GmbH. All rights reserved.

Complete genome of the cellyloytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evloutionary adaptations

Energy Technology Data Exchange (ETDEWEB)

Barabote, Ravi D.; Xie, Gary; Leu, David H.; Normand, Philippe; Necsulea, Anamaria; Daubin, Vincent; Medigue, Claudine; Adney, William S.; Xu,Xin Clare; Lapidus, Alla; Detter, Chris; Pujic, Petar; Bruce, David; Lavire, Celine; Challacombe, Jean F.; Brettin, Thomas S.; Berry, Alison M.

2009-01-01

We present here the complete 2.4 Mb genome of the cellulolytic actinobacterial thermophile, Acidothermus cellulolyticus 11B. New secreted glycoside hydrolases and carbohydrate esterases were identified in the genome, revealing a diverse biomass-degrading enzyme repertoire far greater than previously characterized, and significantly elevating the industrial value of this organism. A sizable fraction of these hydrolytic enzymes break down plant cell walls and the remaining either degrade components in fungal cell walls or metabolize storage carbohydrates such as glycogen and trehalose, implicating the relative importance of these different carbon sources. A novel feature of the A. cellulolyticus secreted cellulolytic and xylanolytic enzymes is that they are fused to multiple tandemly arranged carbohydrate binding modules (CBM), from families 2 and 3. Interestingly, CBM3 was found to be always N-terminal to CBM2, suggesting a functional constraint driving this organization. While the catalytic domains of these modular enzymes are either diverse or unrelated, the CBMs were found to be highly conserved in sequence and may suggest selective substrate-binding interactions. For the most part, thermophilic patterns in the genome and proteome of A. cellulolyticus were weak, which may be reflective of the recent evolutionary history of A. cellulolyticus since its divergence from its closest phylogenetic neighbor Frankia, a mesophilic plant endosymbiont and soil dweller. However, ribosomal proteins and non-coding RNAs (rRNA and tRNAs) in A. cellulolyticus showed thermophilic traits suggesting the importance of adaptation of cellular translational machinery to environmental temperature. Elevated occurrence of IVYWREL amino acids in A. cellulolyticus orthologs compared to mesophiles, and inverse preferences for G and A at the first and third codon positions also point to its ongoing thermoadaptation. Additional interesting features in the genome of this cellulolytic, hot

Cellulolytic enzyme compositions and uses thereof

Energy Technology Data Exchange (ETDEWEB)

Iyer, Prashant; Gaspar, Armindo Ribiero; Croonenberghs, James; Binder, Thomas P.

2017-07-25

The present invention relates enzyme composition comprising a cellulolytic preparation and an acetylxylan esterase (AXE); and the used of cellulolytic enzyme compositions for hydrolyzing acetylated cellulosic material. Finally the invention also relates to processes of producing fermentation products from acetylated cellulosic materials using a cellulolytic enzyme composition of the invention.

Cellulolytic activity of some cellulose-decomposing fungi in salinized soils

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R. A. Badran

2014-08-01

Full Text Available Maximum evolution of CO2 was marked in control soil inoculated by tested fungi but its rate decreased with the increasing salinity. The period of 10 days was most suitable for cellulose degradation by A. niger and P. chrysoecnum and 15 days by A. flavus and C. globosum in control soil. High salinity levels affected greatly the cellulolylic activities of tesled fungi. Carbon content of saline soils increased white the nitrogen content decreased.

Biological hydrogen production by moderately thermophilic anaerobic bacteria

International Nuclear Information System (INIS)

HP Goorissen; AJM Stams

2006-01-01

This study focuses on the biological production of hydrogen at moderate temperatures (65-75 C) by anaerobic bacteria. A survey was made to select the best (moderate) thermophiles for hydrogen production from cellulolytic biomass. From this survey we selected Caldicellulosiruptor saccharolyticus (a gram-positive bacterium) and Thermotoga elfii (a gram-negative bacterium) as potential candidates for biological hydrogen production on mixtures of C 5 -C 6 sugars. Xylose and glucose were used as model substrates to describe growth and hydrogen production from hydrolyzed biomass. Mixed substrate utilization in batch cultures revealed differences in the sequence of substrate consumption and in catabolites repression of the two microorganisms. The regulatory mechanisms of catabolites repression in these microorganisms are not known yet. (authors)

Thermophilic xylanases: from bench to bottle.

Science.gov (United States)

Basit, Abdul; Liu, Junquan; Rahim, Kashif; Jiang, Wei; Lou, Huiqiang

2018-01-17

Lignocellulosic biomass is a valuable raw material. As technology has evolved, industrial interest in new ways to take advantage of this raw material has grown. Biomass is treated with different microbial cells or enzymes under ideal industrial conditions to produce the desired products. Xylanases are the key enzymes that degrade the xylosidic linkages in the xylan backbone of the biomass, and commercial enzymes are categorized into different glycoside hydrolase families. Thermophilic microorganisms are excellent sources of industrially relevant thermostable enzymes that can withstand the harsh conditions of industrial processing. Thermostable xylanases display high-specific activity at elevated temperatures and distinguish themselves in biochemical properties, structures, and modes of action from their mesophilic counterparts. Natural xylanases can be further improved through genetic engineering. Rapid progress with genome editing, writing, and synthetic biological techniques have provided unlimited potential to produce thermophilic xylanases in their natural hosts or cell factories including bacteria, yeasts, and filamentous fungi. This review will discuss the biotechnological potential of xylanases from thermophilic microorganisms and the ways they are being optimized and produced for various industrial applications.

Potential for using thermophilic anaerobic bacteria for bioethanol production from hemicellulose

DEFF Research Database (Denmark)

Sommer, P.; Georgieva, Tania I.; Ahring, Birgitte Kiær

2004-01-01

A limited number of bacteria, yeast and fungi can convert hemicellulose or its monomers (xylose, arabinose, mannose and galactose) into ethanol with a satisfactory yield and productivity. In the present study we tested a number of thermophilic enrichment cultures, and new isolates of thermophilic...... Of D-Xylose into ethanol; (ii) test for viability and ethanol production in pretreated wheat straw hemicellulose hydrolysate; (iii) test for tolerance against high D-xylose concentrations. A total of 86 enrichment cultures and 58 pure cultures were tested and five candidates were selected which...

Anaerobic fungi (phylum Neocallimastigomycota): advances in understanding their taxonomy, life cycle, ecology, role and biotechnological potential.

Science.gov (United States)

Gruninger, Robert J; Puniya, Anil K; Callaghan, Tony M; Edwards, Joan E; Youssef, Noha; Dagar, Sumit S; Fliegerova, Katerina; Griffith, Gareth W; Forster, Robert; Tsang, Adrian; McAllister, Tim; Elshahed, Mostafa S

2014-10-01

Anaerobic fungi (phylum Neocallimastigomycota) inhabit the gastrointestinal tract of mammalian herbivores, where they play an important role in the degradation of plant material. The Neocallimastigomycota represent the earliest diverging lineage of the zoosporic fungi; however, understanding of the relationships of the different taxa (both genera and species) within this phylum is in need of revision. Issues exist with the current approaches used for their identification and classification, and recent evidence suggests the presence of several novel taxa (potential candidate genera) that remain to be characterised. The life cycle and role of anaerobic fungi has been well characterised in the rumen, but not elsewhere in the ruminant alimentary tract. Greater understanding of the 'resistant' phase(s) of their life cycle is needed, as is study of their role and significance in other herbivores. Biotechnological application of anaerobic fungi, and their highly active cellulolytic and hemi-cellulolytic enzymes, has been a rapidly increasing area of research and development in the last decade. The move towards understanding of anaerobic fungi using -omics based (genomic, transcriptomic and proteomic) approaches is starting to yield valuable insights into the unique cellular processes, evolutionary history, metabolic capabilities and adaptations that exist within the Neocallimastigomycota. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Thermostable amylolytic enzymes from a cellulolytic fungus Myceliophthora thermophila D14 (ATCC 48 104)

Energy Technology Data Exchange (ETDEWEB)

Sadhukhan, R K; Manna, S; Roy, S K; Chakrabarty, S L [Bose Research Inst., Calcutta (India). Dept. of Microbiology

1990-09-01

The production of amylolytic enzymes by a thermophilic cellulolytic fungus, Myceliophthora thermophila D14 was investigated by batch cultivation in Czapek-Dox medium at 45deg C. Among various nitrogenous compounds used, NaNO{sub 3} and KNO{sub 3} were found to be the best for amylase production. Starch, cellobiose and maltose induced the synthesis of amylase while glucose, fructose, galactose, lactose, arabinose, xylose, sorbitol, mesoinositol and sucrose did not. Calcium ions had the most stimulating effect on enzyme formation amongst many ions investigated. The synthesis of amylolytic enzymes was dependent on growth and occurred predominantly in the mid-stationary phase. The enzyme was active in a broad temperature range (50deg C-60deg C) and displayed activity optima at 60deg C and pH 5.6. (orig.).

Cellulase enzyme production during continuous culture growth of Sporotrichum (Chrysosporium) thermophile

Energy Technology Data Exchange (ETDEWEB)

Cossar, D; Canevascini, G

1986-07-01

The cellulolytic fungus Sporotrichum (Chrysosporium) thermophile produces an extracellular cellobiose dehydrogenase during batch culture on cellulose or cellobiose. In chemostat culture at pH 5.6 on cellobiose this enzyme was produced in parallel with endo-cellulase. At pH 5.0 in continuous or fed-batch culture such a pattern was not evident. At constant growth rate in a chemostat with varying pH, activity of these enzymes was found to be poorly correlated. Thus the induction of cellobiose dehydrogenase shows a dependence on pH and cellobiose concentration which is different to that for endo-cellulase. The natural inducer of these enzymes and the role of cellubiose dehydrogenase remain to be elucidated.

Interactions of fungi from fermented sausage with regenerated cellulose casings

Science.gov (United States)

Hassan K. Sreenath; Thomas W. Jeffries

2011-01-01

This research examined cellulolytic effects of fungi and other microbes present in cured sausages on the strength and stability of regenerated cellulose casings (RCC) used in the sausage industry. Occasionally during the curing process, RCC would split or fail, thereby leading to loss of product. The fungus Penicillium sp. BT-F-1, which was isolated from fermented...

Cellulolytic (cel) genes of Clostridium thermocellum F7 and the proteins encoded by them

International Nuclear Information System (INIS)

Piruzyan, E.S.; Mogutov, M.A.; Velikodvorskaya, G.A.; Pushkarskaya, T.A.

1988-01-01

This study is concerned with genes cell, ce12, and ce13 encoding the endoglucanase of the cellulolytic complex of the anaerobic thermophilic Clostridium thermocellum F7 bacteria, these genes having been closed by us earlier. The authors present the characteristics of proteins synthesized by the cel genes in the minicell system of the strain Escherichia coli K-12 X925. The molecular weights of the proteins encoded by genes cell, ce12, and ce13 are 30,000, 45,000, and 50,000 dalton, respectively. The study of the homology of the cloned section of the C. thermocellum DNA containing the endoglucanase genes, using Southern's blot-hybridization method, did not reveal their physical linkage in the genome. The authors detected a plasmid with a size of about 30 kb in the cells of the C. thermocellum F7 strain investigated

A comparison of two xylanases from the thermophilic fungi Thielavia terrestris and Thermoascus crustaceus

Energy Technology Data Exchange (ETDEWEB)

Gilbert, M [Ottawa Univ., Dept. of Biology, ON (Canada); Yaguchi, M [Inst. for Biological Sciences, National Research Council of Canada, Ottawa, ON (Canada); Watson, D C [Inst. for Biological Sciences, National Research Council of Canada, Ottawa, ON (Canada); Wong, K K.Y. [Chair of Forest Products Biotechnology, Faculty of Forestry, British Columbia Univ., Vancouver, BC (Canada); Breuil, C [Chair of Forest Products Biotechnology, Faculty of Forestry, British Columbia Univ., Vancouver, BC (Canada); Saddler, J N [Chair of Forest Products Biotechnology, Faculty of Forestry, British Columbia Univ., Vancouver, BC (Canada)

1993-12-01

Two thermophilic xylanases (xylanase II from Thielavia terrestris 255B and the 32-kDa xylanase from Thermoascus crustaceus 235E) were studied to determine if they had different and complementary modes of action when they hydrolysed various types of xylans. Partial amino acid sequencing showed that these two enzymes belonged to different families of [beta]-1,4-glycanases. Xylanase II achieved faster solubilization of insoluble xylan whereas the 32-kDa xylanase was more effective in producing xylose and short xylooligomers. An assessment of the combined hydrolytic action of the two xylanases did not reveal any co-operative action. The sugars released when the two thermophilic xylanases were used together were almost identical to those released when the 32-kDa xylanase acted alone. The two xyalanses were able to remove about 12% of the xylan remaining in an aspen kraft pulp. This indicated that either one of these thermophilic enzymes may be useful for enhancing the bleaching of kraft pulps. (orig.)

Selection of cellulolytic fungi isolated from diverse substrates

Directory of Open Access Journals (Sweden)

Mônica Caramez Triches Damaso

2012-08-01

Full Text Available The aim of the present work was to select filamentous fungi isolated from diverse substrates to obtain the strains with potential to produce the hydrolytic enzymes. From a total of 215 strains, seven strains from the soils, six from the plants and one from sugarcane bagasse were selected and identified as belonging to the Trichoderma, Penicillium and Aspergillus genera. The best hydrolytic activities obtained by semi-solid fermentation using these strains were approximately: 35; 1; 160; 170 and 120 U/gdm (CMCase, FPase, β-glucosidase, xylanase and polygalacturonase, respectively, demonstrating their potential to synthesize the enzymes compared with the results reported in the literature.

Sexual crossing of thermophilic fungus Myceliophthora heterothallica improved enzymatic degradation of sugar beet pulp

NARCIS (Netherlands)

Aguilar-Pontes, Maria Victoria; Zhou, Miaomiao; van der Horst, Sjors; Theelen, Bart; de Vries, Ronald P.; van den Brink, Joost

2016-01-01

Background Enzymatic degradation of plant biomass requires a complex mixture of many different enzymes. Like most fungi, thermophilic Myceliophthora species therefore have a large set of enzymes targeting different linkages in plant polysaccharides. The majority of these enzymes have not been

Evolution of High Cellulolytic Activity in Symbiotic Streptomyces through Selection of Expanded Gene Content and Coordinated Gene Expression

Science.gov (United States)

McDonald, Bradon R.; Takasuka, Taichi E.; Wendt-Pienkowski, Evelyn; Doering, Drew T.; Raffa, Kenneth F.; Fox, Brian G.; Currie, Cameron R.

2016-01-01

The evolution of cellulose degradation was a defining event in the history of life. Without efficient decomposition and recycling, dead plant biomass would quickly accumulate and become inaccessible to terrestrial food webs and the global carbon cycle. On land, the primary drivers of plant biomass deconstruction are fungi and bacteria in the soil or associated with herbivorous eukaryotes. While the ecological importance of plant-decomposing microbes is well established, little is known about the distribution or evolution of cellulolytic activity in any bacterial genus. Here we show that in Streptomyces, a genus of Actinobacteria abundant in soil and symbiotic niches, the ability to rapidly degrade cellulose is largely restricted to two clades of host-associated strains and is not a conserved characteristic of the Streptomyces genus or host-associated strains. Our comparative genomics identify that while plant biomass degrading genes (CAZy) are widespread in Streptomyces, key enzyme families are enriched in highly cellulolytic strains. Transcriptomic analyses demonstrate that cellulolytic strains express a suite of multi-domain CAZy enzymes that are coregulated by the CebR transcriptional regulator. Using targeted gene deletions, we verify the importance of a highly expressed cellulase (GH6 family cellobiohydrolase) and the CebR transcriptional repressor to the cellulolytic phenotype. Evolutionary analyses identify complex genomic modifications that drive plant biomass deconstruction in Streptomyces, including acquisition and selective retention of CAZy genes and transcriptional regulators. Our results suggest that host-associated niches have selected some symbiotic Streptomyces for increased cellulose degrading activity and that symbiotic bacteria are a rich biochemical and enzymatic resource for biotechnology. PMID:27276034

Characterization and Identification of Cellulolytic Bacteria from gut of Worker Macrotermes gilvus

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Andri Ferbiyanto

2015-10-01

Full Text Available As a social insect, termite colony consists of three castes, i.e. reproductive, soldier, and worker castes. In their role of cellulose digestion, the worker termites use two sources of cellulolytic enzyme that include cellulases produced by the termite and the gut symbions. Macrotermes gilvus classified in mound builder termite, mostly depend on cellulolytic bacteria for cellulose digestion. This study aims to characterize cellulolytic bacteria of termite gut symbionts of worker M. gilvus and to identify the cellulolytic bacteria based on sequences of 16S ribosomal RNA (rRNA gene. Cellulolytic bacteria of termite gut were isolated and cultured in CMC (Carboxymethyl cellulose media. The biochemical characters of bacterial isolates were assayed using Microbact 12A and 12B. Cellulolytic activity was determined based on formation of clear zone and cellulolytic index on CMC plate media. The bacterial isolate that has the highest cellulolytic index was analyzed for its 16S rRNA gene sequences. Four isolates of cellulolytic bacteria were successfully isolated from gut of M. gilvus with aerobic and anaerobic conditions. The highest formation of cellulolytic index (2.5 was revealed by RA2. BLAST-N (Basic Local Alignment Search Tool for Nucleotides result of 16S rRNA gene sequences of RU4 and RA2 isolates showed that the isolate has similarity with Bacillus megaterium and Paracoccus yeei, respectively. This result indicated that RA2 isolate was P. yeei, a cellulolytic bacterium of a termite gut of M. gilvus.

Production of cellulases by a thermophilic fungus, Thermoascus aurantiacus A-131

Energy Technology Data Exchange (ETDEWEB)

Kawamori, M; Takayama, K; Takasawa, S

1987-01-01

A thermophilic fungus, strain A-131, isolated from a soil sample produced cellulases in the culture fluid. The fungus (strain A-131) was identified as Thermoascus aurantiacus Miehe from its taxonomical characteristics. The cellulases of T. aurantiacus A-131 were produced constitutively without cellulase inducers. Moreover, their production was induced markedly by amorphous polysaccharides containing beta-1, 4 linkages such as alkali-treated bagasse and xylan rather than crystalline cellulose. The cultivation of T. aurantiacus A-131 at 45 degrees C with 4% alkali-treated bagasse led to the production of about 70 U/ml of CMCase after four days. The thermostability of the cellulolytic enzymes of T. aurantiacus A-131 was excellent and virtually no decreases in their activities were seen after preincubation at 60 degrees C for 24 hours. (Refs. 21).

Cellulase production by white-rot basidiomycetous fungi: solid-state versus submerged cultivation

DEFF Research Database (Denmark)

Bentil, Joseph A.; Thygesen, Anders; Mensah, Moses

2018-01-01

fungi for improved enzyme expression, as well as on-site approaches for production of enzyme blends for industrial biomass conversion. The quantitative comparisons made have implications for selection of the most appropriate cultivation method for WRB fungi for attaining maximal cellulase production....... on different biomass residues in SSC or SmC systems. Although some variation in cellulase production yields have been reported for certain substrates, the analysis convincingly shows that SmC is generally more efficient than SSC for obtaining high cellulase production yields and high cellulase production rates......) or submerged cultivation (SmC) systems. In this review, we quantitatively assess the data available in the literature on cellulase production yields by WRB fungi cultivated by SSC or SmC. The review also assesses cellulolytic enzyme production rates and enzyme recovery when WRB fungi are cultivated...

Ability of some species of fungi of the Basidiomycetes class to degrade cellulose and lignocellulose substrates

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Zdzisław Tagoński

2014-08-01

Full Text Available Studies were carried-out on the ability of 18 strains of 15 white-rot and brown-rot basidiomycetons fungi to degrade wood components and to synthesize cellulolytic enzymes and laccase. 28,5% lignin and 26,1% carbohydrates of pine wood meal, 46,2% lignin and 67,8% carbohydrates of beech wood meal was degraded after 6 weeks incubation by the white-rot fungus Phanerochate chrysosporium. The highest activity of laccase was obtained in from fungi Coriotus zonatus and Fomes fomentarius.

Biochanin A improves fiber fermentation by cellulolytic bacteria

Science.gov (United States)

The objective was to determine the effect of the isoflavone biochanin A (BCA) on rumen cellulolytic bacteria and consequent fermentative activity. When bovine microbial rumen cell suspensions (n = 3) were incubated (24 h, 39 °C) with ground hay, cellulolytic bacteria proliferated, short chain fatty...

Formation of cellulases and degradation of cellulose by several fungi

Energy Technology Data Exchange (ETDEWEB)

Herr, D; Luck, G; Dellweg, H

1978-01-01

Five strains of fungi (Aspergillus niger, Lenzites trabea, Myrothecium verrucaria, Trichoderma koningii and Trichoderma lignorum) were tested for the production of cellulolytic enzymes on pure glucose and on cellulose media. The most active strains belonging to the genera of Trichoderma, Aspergillus and Myrothecium, also secreting high activities of ..beta..-glucosidase, were grown in a bioreactor under defined conditions. Depending on the strain this procedure resulted in a manifold increase in cellulolytic activities. The culture filtrates were concentrated and standardized with respect to ..beta..-glucosidase activity and used for the hydrolysis of cellulose powder. With Trichoderma-cellulase, 46% conversion of crystalline cellulose to glucose was achieved within 48 h. The ratio of cellobiose to glucose found in the hydrolysate, the amount of high molecular carbohydrates as well as the degree of hydrolysis widely depended on the type of cellulase used.

Mesophilic and thermophilic biotreatment of BTEX-polluted air in reactors.

Science.gov (United States)

Mohammad, Balsam T; Veiga, María C; Kennes, Christian

2007-08-15

This study compares the removal of a mixture of benzene, toluene, ethylbenzene, and all three xylene isomers (BTEX) in mesophilic and thermophilic (50 degrees C) bioreactors. In the mesophilic reactor fungi became dominant after long-term operation, while bacteria dominated in the thermophilic unit. Microbial acclimation was achieved by exposing the biofilters to initial BTEX loads of 2-15 g m(-3) h(-1), at an empty bed residence time of 96 s. After adaptation, the elimination capacities ranged from 3 to 188 g m(-3) h(-1), depending on the inlet load, for the mesophilic biofilter with removal efficiencies reaching 96%. On the other hand, in the thermophilic reactor the average removal efficiency was 83% with a maximum elimination capacity of 218 g m(-3) h(-1). There was a clear positive relationship between temperature gradients as well as CO(2) production and elimination capacities across the biofilters. The gas phase was sampled at different depths along the reactors observing that the percentage pollutant removal in each section was strongly dependant on the load applied. The fate of individual alkylbenzene compounds was checked, showing the unusually high biodegradation rate of benzene at high loads under thermophilic conditions (100%) compared to its very low removal in the mesophilic reactor at such load (<10%). Such difference was less pronounced for the other pollutants. After 210 days of operation, the dry biomass content for the mesophilic and thermophilic reactors were 0.300 and 0.114 g g(-1) (support), respectively, reaching higher removals under thermophilic conditions with a lower biomass accumulation, that is, lower pressure drop. (c) 2007 Wiley Periodicals, Inc.

Hydrolytic bacteria in mesophilic and thermophilic degradation of plant biomass

Energy Technology Data Exchange (ETDEWEB)

Zverlov, Vladimir V.; Hiegl, Wolfgang; Koeck, Daniela E.; Koellmeier, Tanja; Schwarz, Wolfgang H. [Department of Microbiology, Technische Universitaet Muenchen, Freising-Weihenstephan (Germany); Kellermann, Josef [Max Planck Institute for Biochemistry, Am Klopferspitz, Martinsried (Germany)

2010-12-15

Adding plant biomass to a biogas reactor, hydrolysis is the first reaction step in the chain of biological events towards methane production. Maize silage was used to enrich efficient hydrolytic bacterial consortia from natural environments under conditions imitating those in a biogas plant. At 55-60 C a more efficient hydrolyzing culture could be isolated than at 37 C. The composition of the optimal thermophilic bacterial consortium was revealed by sequencing clones from a 16S rRNA gene library. A modified PCR-RFLP pre-screening method was used to group the clones. Pure anaerobic cultures were isolated. 70% of the isolates were related to Clostridium thermocellum. A new culture-independent method for identification of cellulolytic enzymes was developed using the isolation of cellulose-binding proteins. MALDI-TOF/TOF analysis and end-sequencing of peptides from prominent protein bands revealed cellulases from the cellulosome of C. thermocellum and from a major cellulase of Clostridium stercorarium. A combined culture of C. thermocellum and C. stercorarium was shown to excellently degrade maize silage. A spore preparation method suitable for inoculation of maize silage and optimal hydrolysis was developed for the thermophilic bacterial consortium. This method allows for concentration and long-term storage of the mixed culture for instance for inoculation of biogas fermenters. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

Genome-Centric Analysis of a Thermophilic and Cellulolytic Bacterial Consortium Derived from Composting

Science.gov (United States)

Lemos, Leandro N.; Pereira, Roberta V.; Quaggio, Ronaldo B.; Martins, Layla F.; Moura, Livia M. S.; da Silva, Amanda R.; Antunes, Luciana P.; da Silva, Aline M.; Setubal, João C.

2017-01-01

Microbial consortia selected from complex lignocellulolytic microbial communities are promising alternatives to deconstruct plant waste, since synergistic action of different enzymes is required for full degradation of plant biomass in biorefining applications. Culture enrichment also facilitates the study of interactions among consortium members, and can be a good source of novel microbial species. Here, we used a sample from a plant waste composting operation in the São Paulo Zoo (Brazil) as inoculum to obtain a thermophilic aerobic consortium enriched through multiple passages at 60°C in carboxymethylcellulose as sole carbon source. The microbial community composition of this consortium was investigated by shotgun metagenomics and genome-centric analysis. Six near-complete (over 90%) genomes were reconstructed. Similarity and phylogenetic analyses show that four of these six genomes are novel, with the following hypothesized identifications: a new Thermobacillus species; the first Bacillus thermozeamaize genome (for which currently only 16S sequences are available) or else the first representative of a new family in the Bacillales order; the first representative of a new genus in the Paenibacillaceae family; and the first representative of a new deep-branching family in the Clostridia class. The reconstructed genomes from known species were identified as Geobacillus thermoglucosidasius and Caldibacillus debilis. The metabolic potential of these recovered genomes based on COG and CAZy analyses show that these genomes encode several glycoside hydrolases (GHs) as well as other genes related to lignocellulose breakdown. The new Thermobacillus species stands out for being the richest in diversity and abundance of GHs, possessing the greatest potential for biomass degradation among the six recovered genomes. We also investigated the presence and activity of the organisms corresponding to these genomes in the composting operation from which the consortium was built

Degradation of natural cellulose by thermophilic and thermotolerant fungi

Energy Technology Data Exchange (ETDEWEB)

Thakre, A P; Johri, B N

1981-06-01

Aspergillus fumigatus Friesen, Sporotrichum sp., Thermoascus aurantiacus Miche and Torula thermophila Cooney and Emerson were able to degrade filter paper to an appreciable extent. Absidia corymbifera (Cohn) Saccardo et Trotter, Rhizopus microsporus van Tieghem and R. rhizopodiformis (Cohn) Zopf could not degrade filter paper though they were able to grow on the media supplied. The fungi able to degrade filter paper were also capable of elaborating extracellular cellulase and produced sufficient hydrolysis of carboxymethyl cellulose. The Cx enzyme was stable at 45 degrees C for 25 days. An incubation period of 15 to 25 days was sufficient not only to degrade CMC but even more complex substrates, such as jute, cotton and filter paper. The cellulases of these fungi were inductive in nature. (Refs. 11).

Stimulation of nitrogen fixation in soddy-podzolic soils with fungi

Science.gov (United States)

Kurakov, A. V.; Prokhorov, I. S.; Kostina, N. V.; Makhova, E. G.; Sadykova, V. S.

2006-09-01

Stimulation of nitrogen fixation in soddy-podzolic soils is related to the hydrolytic activity of fungi decomposing plant polymers. It was found that the rate of nitrogen fixation upon the simultaneous inoculation of the strains of nitrogen-fixing bacteria Bacillus cereus var. mycoides and the cellulolytic fungus Trichoderma asperellum into a sterile soil enriched with cellulose or Jerusalem artichoke residues is two to four times higher than upon the inoculation of the strains of Bacillus cereus var. mycoides L1 only. The increase in the nitrogen fixation depended on the resistance of the substrates added into the soil to fungal hydrolysis. The biomass of the fungi decomposing plant polymers increased by two-four times. The nitrogen-fixing activity of the soil decreased when the growth of the fungi was inhibited with cycloheximide, which attested to a close correlation between the intensity of the nitrogen fixation and the decomposition of the plant polymers by fungi. The introduction of an antifungal antibiotic, together with starch or with plant residues, significantly (by 60-90%) decreased the rate of nitrogen fixation in the soll.

Production of cellulolytic enzymes by fungal cultures. [Aspergillus, Trichoderma, Chaetomium, Stachybotrys, and Hypocrea

Energy Technology Data Exchange (ETDEWEB)

Pyc, R; Fiechter, A. Galas, E.

1977-01-01

Twelve fungal cultures belonging to the genera of Aspergillus, Trichoderma, Chaetomium, Stachybotrys, and Hypocrea were screened for the production of cellulolytic activity. All twelve were found to degrade xylan, avicel, and carboxymethylcellulose. More cellulolytic activity was obtained with shaken cultures than with still cultures and the addition of citrate-phosphate buffer to the media greatly depressed the levels of cellulolytic activity. Varying the composition of the mineral salts in the medium had no effect on the cellulolytic activity. The growth of Aspergillus wentii under controlled conditions in a bioreactor showed that the cellulolytic activity was not affected by the aeration rate or the type of stirrer. The rate of stirring, however, did effect the cellulolytic activity, as at lower stirring speeds considerable wall growth occurred which resulted in low levels of cellulolytic activity. Culture supernatant from Aspergillus wentii was found to hydrolyze from 30-32% of Solka-Floc and from 2-10% of corn cobs, wheat straw, and newsprint. The extensive hydrolysis of Solka-Floc indicates that with suitable treated cellulosic wastes and appropriate enzymes, appreciable amounts of sugars could be obtained.

[Isolation and identification of rumen bacteria for cellulolytic enzyme production].

Science.gov (United States)

Aihemaiti, Maierhaba; Zhen, Fan; Li, Yuezhong; Aibaidoula, Gulisimayi; Yimit, Wusiman

2013-05-04

We screened aerobic bacteria with cellulolytic activity from ruminal fluid of sheep, cattle and camel in Xinjiang. Fresh ruminal fluid was inoculated on sterilized sodium carboxymethylcellulose agar plates. Highly cellulolytic aerobic bacteria were screened out by using Congo red staining and liquid secondary screening culture media. The combination of morphological and biochemical test with 16SrDNA sequence analysis were used to classify the strains. Enzymatic activities of four strains with strong cellulose-decomposing abilities were studied under different culture conditions. Out 84 isolated cellulolytic strains, 40 exhibited strong abilities in decomposing cellulose. They are including 37 Gram-negative isolates and 3 Gram-positive strains. Identification of these 40 strains shows that they belong to 11 species of 6 genera, 16 strains in Stenotrophomonas maltophilia, 10 Ochrobactrum, 5 Sphingobacterium, 3 Microbacterium, 3 Paracoccus and 2 Pseudomonas. The results of the enzymatic studies of four strains with strong cellulolytic abilities indicates that the strains have the best enzyme producing property when straw powder was chosen as the carbon source; the pH at 5.5 -6.0 and temperature at 37 degrees C. The strains with highly cellulolytic abilities isolated from ruminal fluid show strong abilities in cellulose decomposition.

Thermoascus aurantiacus is a promising source of enzymes for biomass deconstruction under thermophilic conditions

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McClendon Shara D

2012-07-01

Full Text Available Abstract Background Thermophilic fungi have attracted increased interest for their ability to secrete enzymes that deconstruct biomass at high temperatures. However, development of thermophilic fungi as enzyme producers for biomass deconstruction has not been thoroughly investigated. Comparing the enzymatic activities of thermophilic fungal strains that grow on targeted biomass feedstocks has the potential to identify promising candidates for strain development. Thielavia terrestris and Thermoascus aurantiacus were chosen for characterization based on literature precedents. Results Thermoascus aurantiacus and Thielavia terrestris were cultivated on various biomass substrates and culture supernatants assayed for glycoside hydrolase activities. Supernatants from both cultures possessed comparable glycoside hydrolase activities when incubated with artificial biomass substrates. In contrast, saccharifications of ionic liquid pretreated switchgrass (Panicum virgatum revealed that T. aurantiacus enzymes released more glucose than T. terrestris enzymes over a range of protein mass loadings and temperatures. Temperature-dependent saccharifications demonstrated that the T. aurantiacus proteins retained higher levels of activity compared to a commercial enzyme mixture sold by Novozymes, Cellic CTec2, at elevated temperatures. Enzymes secreted by T. aurantiacus released glucose at similar protein loadings to CTec2 on dilute acid, ammonia fiber expansion, or ionic liquid pretreated switchgrass. Proteomic analysis of the T. aurantiacus culture supernatant revealed dominant glycoside hydrolases from families 5, 7, 10, and 61, proteins that are key enzymes in commercial cocktails. Conclusions T. aurantiacus produces a complement of secreted proteins capable of higher levels of saccharification of pretreated switchgrass than T. terrestris enzymes. The T. aurantiacus enzymatic cocktail performs at the same level as commercially available enzymatic cocktail for

Thermoascus aurantiacus is a promising source of enzymes for biomass deconstruction under thermophilic conditions.

Science.gov (United States)

McClendon, Shara D; Batth, Tanveer; Petzold, Christopher J; Adams, Paul D; Simmons, Blake A; Singer, Steven W

2012-07-28

Thermophilic fungi have attracted increased interest for their ability to secrete enzymes that deconstruct biomass at high temperatures. However, development of thermophilic fungi as enzyme producers for biomass deconstruction has not been thoroughly investigated. Comparing the enzymatic activities of thermophilic fungal strains that grow on targeted biomass feedstocks has the potential to identify promising candidates for strain development. Thielavia terrestris and Thermoascus aurantiacus were chosen for characterization based on literature precedents. Thermoascus aurantiacus and Thielavia terrestris were cultivated on various biomass substrates and culture supernatants assayed for glycoside hydrolase activities. Supernatants from both cultures possessed comparable glycoside hydrolase activities when incubated with artificial biomass substrates. In contrast, saccharifications of ionic liquid pretreated switchgrass (Panicum virgatum) revealed that T. aurantiacus enzymes released more glucose than T. terrestris enzymes over a range of protein mass loadings and temperatures. Temperature-dependent saccharifications demonstrated that the T. aurantiacus proteins retained higher levels of activity compared to a commercial enzyme mixture sold by Novozymes, Cellic CTec2, at elevated temperatures. Enzymes secreted by T. aurantiacus released glucose at similar protein loadings to CTec2 on dilute acid, ammonia fiber expansion, or ionic liquid pretreated switchgrass. Proteomic analysis of the T. aurantiacus culture supernatant revealed dominant glycoside hydrolases from families 5, 7, 10, and 61, proteins that are key enzymes in commercial cocktails. T. aurantiacus produces a complement of secreted proteins capable of higher levels of saccharification of pretreated switchgrass than T. terrestris enzymes. The T. aurantiacus enzymatic cocktail performs at the same level as commercially available enzymatic cocktail for biomass deconstruction, without strain development or

Biochemical characterization of thermophilic lignocellulose degrading enzymes and their potential for biomass bioprocessing

Energy Technology Data Exchange (ETDEWEB)

Zambare, Vasudeo; Zambare, Archana; Christopher, Lew P. [Center for Bioprocessing Research & Development, South Dakota School of Mines and Technology, Rapid City 57701, SD (United States); Muthukumarappan, Kasiviswanath [Center for Bioprocessing Research & Development, South Dakota State University, Brookings 57007, SD (United States)

2011-07-01

A thermophilic microbial consortium (TMC) producing hydrolytic (cellulolytic and xylanolytic) enzymes was isolated from yard waste compost following enrichment with carboxymethyl cellulose and birchwood xylan. When grown on 5% lignocellulosic substrates (corn stover and prairie cord grass) at 60C, the thermophilic consortium produced more xylanase (up to 489 U/l on corn stover) than cellulase activity (up to 367 U/l on prairie cord grass). Except for the carboxymethyl cellulose-enriched consortium, thermo-mechanical extrusion pretreatment of these substrates had a positive effect on both activities with up to 13% and 21% increase in the xylanase and cellulase production, respectively. The optimum temperatures of the crude cellulase and xylanase were 60C and 70C with half-lives of 15 h and 18 h, respectively, suggesting higher thermostability for the TMC xylanase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the crude enzyme exhibited protein bands of 25-77 kDa with multiple enzyme activities containing 3 cellulases and 3 xylanases. The substrate specificity declined in the following descending order: avicel>birchwood xylan>microcrystalline cellulose>filter paper>pine wood saw dust>carboxymethyl cellulose. The crude enzyme was 77% more active on insoluble than soluble cellulose. The Km and Vmax values were 36.49 mg/ml and 2.98 U/mg protein on avicel (cellulase), and 22.25 mg/ml and 2.09 U/mg protein, on birchwood xylan (xylanase). A total of 50 TMC isolates were screened for cellulase and xylanase secretion on agar plates. All single isolates showed significantly lower enzyme activities when compared to the thermophilic consortia. This is indicative of the strong synergistic interactions that exist within the thermophilic microbial consortium and enhance its hydrolytic capabilities. It was further demonstrated that the thermostable enzyme-generated lignocellulosic hydrolyzates can be fermented to bioethanol by a recombinant strain of Escherichia coli

Compositions for enhancing hydroysis of cellulosic material by cellulolytic enzyme compositions

Science.gov (United States)

Quinlan, Jason; Xu, Feng; Sweeney, Matthew; Johansen, Katja Salomon

2014-09-30

The present invention relates to compositions comprising a GH61 polypeptide having cellulolytic enhancing activity and an organic compound comprising a carboxylic acid moiety, a lactone moiety, a phenolic moiety, a flavonoid moiety, or a combination thereof, wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the organic compound enhances hydrolysis of a cellulosic material by a cellulolytic enzyme compared to the GH61 polypeptide alone or the organic compound alone. The present invention also relates to methods of using the compositions.

An approach to mitigating soil CO2 emission by biochemically inhibiting cellulolytic microbial populations through mediation via the medicinal herb Isatis indigotica

Science.gov (United States)

Wu, Hong-Sheng; Chen, Su-Yun; Li, Ji; Liu, Dong-Yang; Zhou, Ji; Xu, Ya; Shang, Xiao-Xia; Wei, Dong-yang; Yu, Lu-ji; Fang, Xiao-hang; Li, Shun-yi; Wang, Ke-ke

2017-06-01

Greenhouse gases (GHGs, particularly carbon dioxide (CO2)) emissions from soil under wheat production are a significant source of agricultural carbon emissions that have not been mitigated effectively. A field experiment and a static incubation study in a lab were conducted to stimulate wheat growth and investigate its potential to reduce CO2 emissions from soil through intercropping with a traditional Chinese medicinal herb called Isatis indigotica. This work was conducted by adding I. indigotica root exudates based on the quantitative real-time PCR (qPCR) analysis of the DNA copy number of the rhizosphere or bulk soil microbial populations. This addition was performed in relation to the CO2 formation by cellulolytic microorganisms (Penicillium oxalicum, fungi and Ruminococcus albus) to elucidate the microbial ecological basis for the molecular mechanism that decreases CO2 emissions from wheat fields using I. indigotica. The results showed that the panicle weight and full grains per panicle measured through intercropping with I. indigotica (NPKWR) increased by 39% and 28.6%, respectively, compared to that of the CK (NPKW). Intercropping with I. indigotica significantly decreased the CO2 emissions from soil under wheat cultivation. Compared with CK, the total CO2 emission flux during the wheat growth period in the I. indigotica (NPKWR) intercropping treatment decreased by 29.26%. The intensity of CO2 emissions per kg of harvested wheat grain declined from 7.53 kg CO2/kg grain in the NPKW (CK) treatment to 5.55 kg CO2/kg grain in the NPKWR treatment. The qPCR analysis showed that the DNA copy number of the microbial populations of cellulolytic microorganisms (P. oxalicum, fungi and R. albus) in the field rhizosphere around I. indigotica or in the bulk soil under laboratory incubation was significantly lower than that of CK. This finding indicated that root exudates from I. indigotica inhibited the activity and number of cellulolytic microbial populations, which led

Ecological plasticity of Trichoderma fungi in leached chernozem

Science.gov (United States)

Svistova, I. D.; Senchakova, T. Yu.

2010-03-01

The autecological properties of Trichoderma fungi ecotypes isolated from the leached chernozem of the forest-steppe zone of the European part of Russia have been studied. We were the first who carried out the complex study of the synecological relations of micromycetes of such kinds in a system including the soil, microbial community, and plants, i.e., their relations with soil saprotrophic fungi, bacteria, actinomycetes, plants, and pathogenic fungi. It was shown that the ecological plasticity of the Trichoderma genus in the soil of this zone is determined by its growth rate, the optimum pH and temperature, the biosynthesis of extracellular hydrolytic enzymes, the biological action of mycotoxins, and the ability for parasitism. The efficiency of the introduction of Trichoderma species typical and atypical for the leached chernozem into this soil and their influence on the structure of the microbial community were evaluated. The T. pseudokoningii ecotype, which produces cellulolytic enzymes, is very promising for industrial biotechnology, and the T. harzianum ecotype can be used in soil biotechnology for the biocontrol of chernozem. The addition of a commercial trichodermin preparation into the chernozem damages the structure of its microbial community.

Enzymatic saccharification of biologically pre-treated wheat straw with white-rot fungi.

Science.gov (United States)

Dias, Albino A; Freitas, Gil S; Marques, Guilhermina S M; Sampaio, Ana; Fraga, Irene S; Rodrigues, Miguel A M; Evtuguin, Dmitry V; Bezerra, Rui M F

2010-08-01

Wheat straw was submitted to a pre-treatment by the basidiomycetous fungi Euc-1 and Irpex lacteus, aiming to improve the accessibility of cellulose towards enzymatic hydrolysis via previous selective bio-delignification. This allowed the increase of substrate saccharification nearly four and three times while applying the basidiomycetes Euc-1 and I. lacteus, respectively. The cellulose/lignin ratio increased from 2.7 in the untreated wheat straw to 5.9 and 4.6 after the bio-treatment by the basidiomycetes Euc-1 and I. lacteus, respectively, thus evidencing the highly selective lignin biodegradation. The enzymatic profile of both fungi upon bio-treatment of wheat straw have been assessed including laccase, manganese-dependent peroxidase, lignin peroxidase, carboxymethylcellulase, xylanase, avicelase and feruloyl esterase activities. The difference in efficiency and selectivity of delignification within the two fungi treatments was interpreted in terms of specific lignolytic enzyme profiles and moderate xylanase and cellulolytic activities. (c) 2010 Elsevier Ltd. All rights reserved.

Cellulolytic and xylanolytic potential of high β-glucosidase-producing Trichoderma from decaying biomass.

Science.gov (United States)

Okeke, Benedict C

2014-10-01

Availability, cost, and efficiency of microbial enzymes for lignocellulose bioconversion are central to sustainable biomass ethanol technology. Fungi enriched from decaying biomass and surface soil mixture displayed an array of strong cellulolytic and xylanolytic activities. Strains SG2 and SG4 produced a promising array of cellulolytic and xylanolytic enzymes including β-glucosidase, usually low in cultures of Trichoderma species. Nucleotide sequence analysis of internal transcribed spacer 2 (ITS2) region of rRNA gene revealed that strains SG2 and SG4 are closely related to Trichoderma inhamatum, Trichoderma piluliferum, and Trichoderma aureoviride. Trichoderma sp. SG2 crude culture supernatant correspondingly displayed as much as 9.84 ± 1.12, 48.02 ± 2.53, and 30.10 ± 1.11 units mL(-1) of cellulase, xylanase, and β-glucosidase in 30 min assay. Ten times dilution of culture supernatant of strain SG2 revealed that total activities were about 5.34, 8.45, and 2.05 orders of magnitude higher than observed in crude culture filtrate for cellulase, xylanase, and β-glucosidase, respectively, indicating that more enzymes are present to contact with substrates in biomass saccharification. In parallel experiments, Trichoderma species SG2 and SG4 produced more β-glucosidase than the industrial strain Trichoderma reesei RUT-C30. Results indicate that strains SG2 and SG4 have potential for low cost in-house production of primary lignocellulose-hydrolyzing enzymes for production of biomass saccharides and biofuel in the field.

Isolation of microbial native Stumps with cellulolytic activity of a compost process

International Nuclear Information System (INIS)

Jaramillo G, Marisol; Ruiz V Orlando Simon; Yepes P, Maria del Socorro; Montoya C, Olga Ines

2003-01-01

The isolation, selection adaptation and handling of native microorganisms coming from organic waste are an alternative to avoid the accumulation and the lack of the proper use of these undesirable materials. This organic waste is a source for obtaining microbial strains, which are potentially producers of Industrial enzymes and, at the same time, it works as substrate so that these organisms can transform it into compost or organic manure. In this work, 39 native strains of microorganisms with potential cellulolytic activity coming from the organic waste of the urban and rural sector, from the Compost Plant of Marinilla Antioquia) municipality, were isolated, evaluated and purified. The waste was previously selected and then submitted to an aerobic degradation or compost. The microbial strains were isolated in a selective medium with carboxymethyl cellulose (CMC), of the phases mesophile, termophile, cooling and maturation of the compost process. Eighty-two percent (82%)of the obtained colonies were identified, in principle as Bacillus, because of their morphology and their reaction to the Gram coloration. The fungi population was seen only during the cooling phase. Then, the potential cellulolytic activity was evaluated qualitatively in a solid medium with the Congo Red coloration, with which the Beta-endoglucanase activity was evaluated through the formation of clarified zones. Such staining was applied in two mediums with CMC with and without glucose It was observed that 33.3% of the isolated organisms produced the enzyme In both mediums; however, 25.6% of microorganisms did not show the production of this enzyme, and only 15.8% did not require the inducers to produce it

Physiology of thermophilic bacteria

Energy Technology Data Exchange (ETDEWEB)

Ljungdahl, L G

1979-01-01

Thermophilic micro-organisms have all of the properties normally found in mesophilic micro-organisms. These include metabolic pathways, regulatory mechanisms such as allosteric or feedback control, repression and induction of protein synthesis, growth yields and metabolic rates. The main difference between thermophiles and mesophiles is the former's capacity to grow at high temperatures. The basis for this capacity is the thermophile's capability to synthesize proteins, complex structures and membranes that are stable or are stabilized and functional at thermophilic temperatures. It is proposed that the maximum and minimum growth temperatures are normally determined by properties associated with proteins, and that the membrane plays a lesser role in determining these temperatures. Enzymes and other proteins from thermophiles, except for having higher thermostability, are very similar to corresponding proteins from mesophiles. The higher thermostability is generally dependent on subtle changes in the composition and sequence of the amino acids and rarely dependent on non-proteinaceous factors. Although over 100 proteins have been purified from thermophiles and compared with corresponding proteins from mesophiles, the exact nature of the higher thermostability has yet to be determined in a protein from a thermophile.

Properties of thermophilic microorganisms

International Nuclear Information System (INIS)

Ljungdahl, L.G.

1984-01-01

Microorganisms are called thermophilic or extreme thermophilic (caldo-active) if they grow and reproduce over 47 0 C and 70 0 C, respectively. A survey of growth characteristics of thermophiles is presented and it includes those which also live at extreme pH. The prevalent but not completely emcompassing theory of the ability of thermophiles to grow at high temperatures is that they have macromolecules and cell organelles with high thermostability. Work on some proteins and cell organelles from thermophiles is reviewed. The thermostabilities of these components are compared with those of the living cells, and factors which may govern optimum as well as minimum growth temperatures of microorganisms are discussed. Examples are from the literature but also include enzymes involved in tetrahydrofolate metabolism and other proteins of acetogenic therhmophilic bacteria which are presently studied in the author's laboratory

Microorganisms applying for artificial soil regeneration technology in space greenhouses

Science.gov (United States)

Krivobok, A. S.

2012-04-01

The space greenhouse and technology for growing plants are being designed in frame of bio-technical life support systems development. During long-term space missions such greenhouse could provide the crew with vitamins and rough plant fiber. One of the important elements of the plant cultivation technology in the absence of earth gravity is organization and support the optimum root area. The capillary-porous substrate composed of anionites (FIBAN -1) and cationites (FIBAN -22-1) synthetic salt-saturated fibers is developed for plant cultivation in space and named "BIONA-V3". The BIONA main features are high productivity and usability. But the pointed features are not constant: the substrate productivity will be decreasing gradually from vegetation to vegetation course of plant residues and root secretions accumulation. Also, the basic hydro-physical characteristic of root zone will be shifted. Furthermore, saprotrophic microflora will develop and lead to increasing the level of microbial contamination of whole inhabit isolated module. Due to these changes the substrate useful life is limited and store mass is increased in long-term missions. For overhaul-period renewal it' necessary to remove the roots residues and other organic accumulation providing safety of the substrate capillary-porous structure. The basic components of 24-days old plant roots (Brassica chinensis, L) are cellulose (35 %) hemicellulose (11 %) and lignin (10 %). We see that one of the possible ways for roots residues removal from fibrous BIONA is microorganisms applying with strong cellulolytic and ligninolytic activities. The fungi Trichoderma sp., cellulolytic bacteria associations, and some genus of anaerobic thermophilic cellulolitic bacteria have been used for roots residues biodegradation. In case of applying cellulolytic fungi Trichoderma sp. considerable decrease of microcrystalline cellulose has been noted in both liquid and solid state fermentation. Cellulolytic fungi weight has been

Insight into Dominant Cellulolytic Bacteria from Two Biogas Digesters and Their Glycoside Hydrolase Genes

Science.gov (United States)

Zhang, Jun; Zhang, Lei; Geng, Alei; Liu, Fanghua; Zhao, Guoping; Wang, Shengyue; Zhou, Zhihua; Yan, Xing

2015-01-01

Diverse cellulolytic bacteria are essential for maintaining high lignocellulose degradation ability in biogas digesters. However, little was known about functional genes and gene clusters of dominant cellulolytic bacteria in biogas digesters. This is the foundation to understand lignocellulose degradation mechanisms of biogas digesters and apply these gene resource for optimizing biofuel production. A combination of metagenomic and 16S rRNA gene clone library methods was used to investigate the dominant cellulolytic bacteria and their glycoside hydrolase (GH) genes in two biogas digesters. The 16S rRNA gene analysis revealed that the dominant cellulolytic bacteria were strains closely related to Clostridium straminisolvens and an uncultured cellulolytic bacterium designated BG-1. To recover GH genes from cellulolytic bacteria in general, and BG-1 in particular, a refined assembly approach developed in this study was used to assemble GH genes from metagenomic reads; 163 GH-containing contigs ≥ 1 kb in length were obtained. Six recovered GH5 genes that were expressed in E. coli demonstrated multiple lignocellulase activities and one had high mannanase activity (1255 U/mg). Eleven fosmid clones harboring the recovered GH-containing contigs were sequenced and assembled into 10 fosmid contigs. The composition of GH genes in the 163 assembled metagenomic contigs and 10 fosmid contigs indicated that diverse GHs and lignocellulose degradation mechanisms were present in the biogas digesters. In particular, a small portion of BG-1 genome information was recovered by PhyloPythiaS analysis. The lignocellulase gene clusters in BG-1 suggested that it might use a possible novel lignocellulose degradation mechanism to efficiently degrade lignocellulose. Dominant cellulolytic bacteria of biogas digester possess diverse GH genes, not only in sequences but also in their functions, which may be applied for production of biofuel in the future. PMID:26070087

Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen.

Science.gov (United States)

Denman, Stuart E; McSweeney, Christopher S

2006-12-01

Traditional methods for enumerating and identifying microbial populations within the rumen can be time consuming and cumbersome. Methods that involve culturing and microscopy can also be inconclusive, particularly when studying anaerobic rumen fungi. A real-time PCR SYBR Green assay, using PCR primers to target total rumen fungi and the cellulolytic bacteria Ruminococcus flavefaciens and Fibrobacter succinogenes, is described, including design and validation. The DNA and crude protein contents with respect to the fungal biomass of both polycentric and monocentric fungal isolates were investigated across the fungal growth stages to aid in standard curve generation. The primer sets used were found to be target specific with no detectable cross-reactivity. Subsequently, the real-time PCR assay was employed in a study to detect these populations within cattle rumen. The anaerobic fungal target was observed to increase 3.6-fold from 0 to 12 h after feeding. The results also indicated a 5.4-fold increase in F. succinogenes target between 0 and 12 h after feeding, whereas R. flavefaciens was observed to maintain more or less consistent levels. This is the first report of a real-time PCR assay to estimate the rumen anaerobic fungal population.

Thermophilic lignocellulose deconstruction.

Science.gov (United States)

Blumer-Schuette, Sara E; Brown, Steven D; Sander, Kyle B; Bayer, Edward A; Kataeva, Irina; Zurawski, Jeffrey V; Conway, Jonathan M; Adams, Michael W W; Kelly, Robert M

2014-05-01

Thermophilic microorganisms are attractive candidates for conversion of lignocellulose to biofuels because they produce robust, effective, carbohydrate-degrading enzymes and survive under harsh bioprocessing conditions that reflect their natural biotopes. However, no naturally occurring thermophile is known that can convert plant biomass into a liquid biofuel at rates, yields and titers that meet current bioprocessing and economic targets. Meeting those targets requires either metabolically engineering solventogenic thermophiles with additional biomass-deconstruction enzymes or engineering plant biomass degraders to produce a liquid biofuel. Thermostable enzymes from microorganisms isolated from diverse environments can serve as genetic reservoirs for both efforts. Because of the sheer number of enzymes that are required to hydrolyze plant biomass to fermentable oligosaccharides, the latter strategy appears to be the preferred route and thus has received the most attention to date. Thermophilic plant biomass degraders fall into one of two categories: cellulosomal (i.e. multienzyme complexes) and noncellulosomal (i.e. 'free' enzyme systems). Plant-biomass-deconstructing thermophilic bacteria from the genera Clostridium (cellulosomal) and Caldicellulosiruptor (noncellulosomal), which have potential as metabolic engineering platforms for producing biofuels, are compared and contrasted from a systems biology perspective. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Bioconversion process of rice straw by thermotolerant cellulolytic ...

African Journals Online (AJOL)

Administrator

2011-09-26

state fermentation for bioethanol production is a focus of current attention. ... Optimization of fermentation conditions showed highest cellulolytic enzymes ... using dilute acid pretreated rice straw hydrolysate with initial soluble ...

Malbranchea cinnamomea: A thermophilic fungal source of catalytically efficient lignocellulolytic glycosyl hydrolases and metal dependent enzymes.

Science.gov (United States)

Mahajan, Chhavi; Basotra, Neha; Singh, Surender; Di Falco, Marcos; Tsang, Adrian; Chadha, B S

2016-01-01

This study reports thermophilic fungus Malbranchea cinnamomea as an important source of lignocellulolytic enzymes. The secretome analysis using LC-MS/MS orbitrap showed that fungus produced a spectrum of glycosyl hydrolases (cellulase/hemicellulase), polysaccharide lyases (PL) and carbohydrate esterases (CE) in addition to cellobiose dehydrogenase (CDH) indicating the presence of functional classical and oxidative cellulolytic mechanisms. The protein fractions in the secretome resolved by ion exchange chromatography were analyzed for ability to hydrolyze alkali treated carrot grass (ATCG) in the presence of Mn(2+)/Cu(2+). This strategy in tandem with peptide mass fingerprinting led to identification of metal dependent protein hydrolases with no apparent hydrolytic activity, however, showed 5.7 folds higher saccharification in presence of Mn(2+). Furthermore, adding different protein fractions to commercial cellulase (Novozymes: Cellic CTec2) resulted in enhanced hydrolysis of ATCG ranging between 1.57 and 3.43 folds indicating the enzymes from M. cinnamomea as catalytically efficient. Copyright © 2015 Elsevier Ltd. All rights reserved.

Thermophilic Biohydrogen Production

DEFF Research Database (Denmark)

Karakashev, Dimitar Borisov; Angelidaki, Irini

2011-01-01

Dark fermentative hydrogen production at thermophilic conditions is attractive process for biofuel production. From thermodynamic point of view, higher temperatures favor biohydrogen production. Highest hydrogen yields are always associated with acetate, or with mixed acetate- butyrate type...... fermentation. On the contrary the hydrogen yield decreases, with increasing concentrations of lactate, ethanol or propionate. Major factors affecting dark fermentative biohydrogen production are organic loading rate (OLR), pH, hydraulic retention time (HRT), dissolved hydrogen and dissolved carbon dioxide...... concentrations, and soluble metabolic profile (SMP). A number of thermophilic and extreme thermophilic cultures (pure and mixed) have been studied for biohydrogen production from different feedstocks - pure substrates and waste/wastewaters. Variety of process technologies (operational conditions...

Genome sequence and transcriptome analyses of the thermophilic zygomycete fungus Rhizomucor miehei.

Science.gov (United States)

Zhou, Peng; Zhang, Guoqiang; Chen, Shangwu; Jiang, Zhengqiang; Tang, Yanbin; Henrissat, Bernard; Yan, Qiaojuan; Yang, Shaoqing; Chen, Chin-Fu; Zhang, Bing; Du, Zhenglin

2014-04-21

The zygomycete fungi like Rhizomucor miehei have been extensively exploited for the production of various enzymes. As a thermophilic fungus, R. miehei is capable of growing at temperatures that approach the upper limits for all eukaryotes. To date, over hundreds of fungal genomes are publicly available. However, Zygomycetes have been rarely investigated both genetically and genomically. Here, we report the genome of R. miehei CAU432 to explore the thermostable enzymatic repertoire of this fungus. The assembled genome size is 27.6-million-base (Mb) with 10,345 predicted protein-coding genes. Even being thermophilic, the G + C contents of fungal whole genome (43.8%) and coding genes (47.4%) are less than 50%. Phylogenetically, R. miehei is more closerly related to Phycomyces blakesleeanus than to Mucor circinelloides and Rhizopus oryzae. The genome of R. miehei harbors a large number of genes encoding secreted proteases, which is consistent with the characteristics of R. miehei being a rich producer of proteases. The transcriptome profile of R. miehei showed that the genes responsible for degrading starch, glucan, protein and lipid were highly expressed. The genome information of R. miehei will facilitate future studies to better understand the mechanisms of fungal thermophilic adaptation and the exploring of the potential of R. miehei in industrial-scale production of thermostable enzymes. Based on the existence of a large repertoire of amylolytic, proteolytic and lipolytic genes in the genome, R. miehei has potential in the production of a variety of such enzymes.

Improved cellulose conversion to bio-hydrogen with thermophilic bacteria and characterization of microbial community in continuous bioreactor

International Nuclear Information System (INIS)

Jiang, Hongyu; Gadow, Samir I.; Tanaka, Yasumitsu; Cheng, Jun; Li, Yu-You

2015-01-01

Thermophilic hydrogen fermentation of cellulose was evaluated by a long term continuous experiment and batch experiments. The continuous experiment was conducted under 55 °C using a continuously stirred tank reactor (CSTR) at a hydraulic retention time (HRT) of 10 day. A stable hydrogen yield of 15.4 ± 0.23 mol kg −1 of cellulose consumed was maintained for 190 days with acetate and butyrate as the main soluble byproducts. An analysis of the 16S rRNA sequences showed that the hydrogen-producing thermophilic cellulolytic microorganisms (HPTCM) were close to Thermoanaerobacterium thermosaccharolyticum, Clostridium sp. and Enterobacter cloacae. Batch experiment demonstrated that the highest H 2 producing activity was obtained at 55 °C and the ultimate hydrogen yield and the metabolic by-products were influenced greatly by temperatures. The effect of temperature variation showed that the activation energy for cellulose and glucose were estimated at 103 and 98.8 kJ mol −1 , respectively. - Highlights: • Continuous cellulosic-hydrogen fermentation was conducted at 55 °C. • Hydrogen yield was improved to 15.4 mol kg −1 of consumed-cellulose. • The cellulosic hydrogen bacteria were close to Clostridia and Enterobacter genus. • The mixed microflora produced H 2 within a wide range of temperatures (35 °C–65 °C). • Activation energy of cellulose and glucose were 103 and 98.8 kJ mol −1 , respectively

Composition of cellulase complex of Clostridium thermocellum

Energy Technology Data Exchange (ETDEWEB)

Golovchenko, N P; Chuvil' skaya, N A; Akimenko, V K

1985-01-01

It is thought that the anaerobic thermophilic cellulolytic bacterium C. thermocellum has the potential for direct industrial bioconversion of cellulose into ethanol. Therefore, much attention has been given to the study of the cellulolytic properties of the culture and to the characteristics of the cellulose complex, which is still not completely understood. Hence, the activity and location of various cellulolytic enzymes of C. thermocellum were determined. C. thermocellum has 6 known cellulolytic enzymes. Endoglucanase, cellobiohydrolase and exoglucosidase are extracellular enzymes (99-100 percent of the activity is located outside the cells) while cellulobiases, cellobiose phosphorylase and cellodextrine phosphorylase are inside the cells (80-90% of the activity). 25 references.

Methanogenesis in Thermophilic Biogas Reactors

DEFF Research Database (Denmark)

Ahring, Birgitte Kiær

1995-01-01

Methanogenesis in thermophilic biogas reactors fed with different wastes is examined. The specific methanogenic activity with acetate or hydrogen as substrate reflected the organic loading of the specific reactor examined. Increasing the loading of thermophilic reactors stabilized the process as ....... Experiments using biogas reactors fed with cow manure showed that the same biogas yield found at 550 C could be obtained at 610 C after a long adaptation period. However, propionate degradation was inhibited by increasing the temperature.......Methanogenesis in thermophilic biogas reactors fed with different wastes is examined. The specific methanogenic activity with acetate or hydrogen as substrate reflected the organic loading of the specific reactor examined. Increasing the loading of thermophilic reactors stabilized the process...... as indicated by a lower concentration of volatile fatty acids in the effluent from the reactors. The specific methanogenic activity in a thermophilic pilot-plant biogas reactor fed with a mixture of cow and pig manure reflected the stability of the reactor. The numbers of methanogens counted by the most...

Biosynthesis of cellulolytic enzymes by Tricothecium roseum with ...

African Journals Online (AJOL)

STORAGESEVER

2008-11-05

Nov 5, 2008 ... good inducer for extracellular cellulolytic enzyme production by the fungus. Key words: Tricothecium ... feasible for the conversion of cellulose into fermentable sugars and fuel ... Biomass of the culture was dried at 70°C in an ...

Anaerobic solid state fermentation of cellulosic substrates with possible application to cellulase production

Energy Technology Data Exchange (ETDEWEB)

Vandevoorde, L; Verstraete, W

1987-08-01

A solid state fermentation process was developed for the conversion of straw and cellulose under anaerobic conditions by a mixed culture of cellulolytic and methanogenic organisms. The bioconversion rate and efficiency were compared under mesophilic (35/sup 0/C) and thermophilic (55/sup 0/C) conditions. Cellulolytic activity was assayed in terms of sugar and overall soluble organic matter (chemical oxygen demand, COD) production. Maximum conversion rates were obtained under thermophilic conditions, i.e. 8.4 g and 14.2 g COD/kg.d, respectively, when wheat straw and cellulose were used as substrates. The cellulolytic activity of the reactor contents (23% dry matter), measured under substrate excess conditions, amounted to 50 g COD/kg.d. As a comparison, the activity of rumen contents (15 % dry matter) measured by the same assay amounted to 150 g COD/kg . d. The anaerobic cellulases appeared to be substrate bound. This and the relative low activity levels attained, limit the perspectives of producing cellulase enzymes by this type of process.

Biohydrogen Production by the Thermophilic Bacterium Caldicellulosiruptor saccharolyticus: Current Status and Perspectives

Directory of Open Access Journals (Sweden)

Servé W. M. Kengen

2013-01-01

Full Text Available Caldicellulosiruptor saccharolyticus is one of the most thermophilic cellulolytic organisms known to date. This Gram-positive anaerobic bacterium ferments a broad spectrum of mono-, di- and polysaccharides to mainly acetate, CO2 and hydrogen. With hydrogen yields approaching the theoretical limit for dark fermentation of 4 mol hydrogen per mol hexose, this organism has proven itself to be an excellent candidate for biological hydrogen production. This review provides an overview of the research on C. saccharolyticus with respect to the hydrolytic capability, sugar metabolism, hydrogen formation, mechanisms involved in hydrogen inhibition, and the regulation of the redox and carbon metabolism. Analysis of currently available fermentation data reveal decreased hydrogen yields under non-ideal cultivation conditions, which are mainly associated with the accumulation of hydrogen in the liquid phase. Thermodynamic considerations concerning the reactions involved in hydrogen formation are discussed with respect to the dissolved hydrogen concentration. Novel cultivation data demonstrate the sensitivity of C. saccharolyticus to increased hydrogen levels regarding substrate load and nitrogen limitation. In addition, special attention is given to the rhamnose metabolism, which represents an unusual type of redox balancing. Finally, several approaches are suggested to improve biohydrogen production by C. saccharolyticus.

Isolation and identification of cellulolytic bacteria from termites gut (Cryptotermes sp.)

Science.gov (United States)

Peristiwati; Natamihardja, Y. S.; Herlini, H.

2018-05-01

The energy and environmental crises developed due to a huge amount of cellulosic materials are disposed of as “waste.” Cellulose is the most abundant biopolymer on Earth. The hydrolysis of cellulose to glucose and soluble sugars has thus become a subject of intense research. Termites are one of the most important soil insects that efficiently decompose lignocelluloses with the aid of their associated microbial symbionts to a simpler form of sugars. The steps of this study consisted of cellulose isolation, cellulolytic bacteria isolation and identification. Cellulose degrading bacteria from termite (Cryptotermes sp.) gut flora were isolated, screened and their identification was studied which showed halo zones due to CMC agar. Among 12 isolates of bacteria, six isolates were cellulolytic. MLC-A isolate had shown a maximum in a cellulolytic index (1.32). Each isolate was identified based on standard physical and biochemical tests. Three isolates were identified in the genus of Clostridium, one isolate be placed in the group of Mycobacteriaceae, Lactobacillaceae or Coryneform and the last one in the genus Proteus.

Utilization of steam- and explosion-decompressed aspen wood by some anaerobes

Energy Technology Data Exchange (ETDEWEB)

Khan, A W; Asther, M; Giuliano, C

1984-01-01

Tests made to study the suitability of using steam- and explosion-decompressed aspen wood as a substrate in anaerobic fermentations indicated that after washing with dilute NaOH it becomes less than 80% accessible to both mesophilic and thermophilic cellulolytic anaerobes and cellulases, compared with delignified, ball-milled pulp. After washing, this material was also suitable for the single-step conversion of cellulose to EtOH using cocultures consisting of cellulolytic and EtOH-producing saccharolytic anaerobes; and without and after washing by the use of cellulolytic enzymes and ethanologenic anaerobes.

Characterization of Cellulolytic Bacterial Cultures Grown in Different Substrates

Directory of Open Access Journals (Sweden)

Mohamed Idris Alshelmani

2013-01-01

Full Text Available Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ and the American Type Culture Collection (ATCC. The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF of palm kernel cake (PKC. The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30∘C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC.

An innovative approach for hyperproduction of cellulolytic and ...

African Journals Online (AJOL)

The present work describes the production of cellulolytic enzymes and hemicellulolytic enzyme (xylanase) along with total extracellular protein by Aspergillus niger and Trichoderma viride using submerged fermentation. Among seven different kinds of experiments, secretion rate of protein and enzymes was investigated by ...

EVALUATION OF ENDOGLUCANASE, EXOGLUCANASE, LACCASE, AND LIGNIN PEROXIDASE ACTIVITIES ON TEN WHITE-ROT FUNGI

Directory of Open Access Journals (Sweden)

Sandra Montoya B

2014-12-01

Full Text Available This paper presents a way of tracking the production of lignocellulolytic enzymes in ten species of white rot fungi: Lentinula edodes, Schizophyllum commune, Trametes trogii, Coriolus versicolor, Pycnoporus sanguineus, Ganoderma applanatum, Ganoderma lucidum, Grifola frondosa, Pleurotus ostreatus and Auricularia delicata. These species were first screened on solid culture media containing carboxymethyl cellulose, crystalline cellulose, ABTS (2,2´-azino-bis(3-ethylbenzothiazoline-6-sulphonate and azure B, which showed the production of endoglucanase, exoglucanase, laccase and lignin peroxidase (LiP enzymes. Cellulolytic activities were detected after five days of incubation with congo red indicator, forming a clear-white halo in areas where cellulose was degraded. For ligninases, the tracking consisted of the monitoring in the formation of green halos due to ABTS oxidation for laccase, and decolorization halos on azure B for LiP during 14 days of incubation. From this qualitative screening, four strains were selected (G. lucidum, L. edodes, C. versicolor and T. trogii as the best producers of cellulolytic and ligninolytic enzymes. These four species were inoculated on a substrate of sawdust oak, yielding 51,8% of lignin degraded by L. edodes and 22% of cellulose degraded by C. versicolor.

Isolation and Identification of cellulolytic bacteria from mangrove sediment in Bangka Island

Science.gov (United States)

Kurniawan, A.; Prihanto, A. A.; Sari, S. P.; Febriyanti, D.; Kurniawan, A.; Sambah, A. B.; Asriani, E.

2018-04-01

Cellulolytic bacteria is bacteria which hydrolyze cellulose to reducing sugars. This research aims to obtain cellulolytic bacteria from the sediment of mangroves in Bangka island. Reasearch was conducted from March to August 2017. Sampling was conducted at Sungailiat, and Tukak Sadai, South of Bangka. Bacteria was isolated using 1% Carboxymetyl Cellulosa (CMC). The isolation resulted in four isolates from Sungailiat and nine isolates from Tukak Sadai. Total five isolates, namely Bacillus pumilus, Pseudomonas sp., Bacillus amyloliquefacien, Bacillus alvei, Bacillus coagulant were identified. The best isolates that produced cellulose was Pseudomonas aeruginosa.

Functional diversity of family 3 β-glucosidases from thermophilic cellulolytic fungus Humicola insolens Y1.

Science.gov (United States)

Xia, Wei; Bai, Yingguo; Cui, Ying; Xu, Xinxin; Qian, Lichun; Shi, Pengjun; Zhang, Wei; Luo, Huiying; Zhan, Xiuan; Yao, Bin

2016-06-08

The fungus Humicola insolens is one of the most powerful decomposers of crystalline cellulose. However, studies on the β-glucosidases from this fungus remain insufficient, especially on glycosyl hydrolase family 3 enzymes. In the present study, we analyzed the functional diversity of three distant family 3 β-glucosidases from Humicola insolens strain Y1, which belonged to different evolutionary clades, by heterogeneous expression in Pichia pastoris strain GS115. The recombinant enzymes shared similar enzymatic properties including thermophilic and neutral optima (50-60 °C and pH 5.5-6.0) and high glucose tolerance, but differed in substrate specificities and kinetics. HiBgl3B was solely active towards aryl β-glucosides while HiBgl3A and HiBgl3C showed broad substrate specificities including both disaccharides and aryl β-glucosides. Of the three enzymes, HiBgl3C exhibited the highest specific activity (158.8 U/mg on pNPG and 56.4 U/mg on cellobiose) and catalytic efficiency and had the capacity to promote cellulose degradation. Substitutions of three key residues Ile48, Ile278 and Thr484 of HiBgl3B to the corresponding residues of HiBgl3A conferred the enzyme activity towards sophorose, and vice versa. This study reveals the functional diversity of GH3 β-glucosidases as well as the key residues in recognizing +1 subsite of different substrates.

ENDOSPORES OF THERMOPHILIC FERMENTATIVE BACTERIA

DEFF Research Database (Denmark)

Volpi, Marta

2016-01-01

solely based on endospores of sulphate-reducing bacteria (SRB), which presumably constitute only a small fraction of the total thermophilic endospore community reaching cold environments. My PhD project developed an experimental framework for using thermophilic fermentative endospores (TFEs) to trace...

Hydrogen Production by Thermophilic Fermentation

NARCIS (Netherlands)

Niel, van E.W.J.; Willquist, K.; Zeidan, A.A.; Vrije, de T.; Mars, A.E.; Claassen, P.A.M.

2012-01-01

Of the many ways hydrogen can be produced, this chapter focuses on biological hydrogen production by thermophilic bacteria and archaea in dark fermentations. The thermophiles are held as promising candidates for a cost-effective fermentation process, because of their relatively high yields and broad

Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

Science.gov (United States)

Maiyuran, Suchindra; Kramer, Randall; Harris, Paul

2013-10-29

The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Myceliophthora thermophila syn. Sporotrichum thermophile: a thermophilic mould of biotechnological potential.

Science.gov (United States)

Singh, Bijender

2016-01-01

Myceliophthora thermophila syn. Sporotrichum thermophile is a ubiquitous thermophilic mould with a strong ability to degrade organic matter during optimal growth at 45 °C. Both genome analysis and experimental data have suggested that the mould is capable of hydrolyzing all major polysaccharides found in biomass. The mould is able to secrete a large number of hydrolytic enzymes (cellulases, laccases, xylanases, pectinases, lipases, phytases and some other miscellaneous enzymes) employed in various biotechnological applications. Characterization of the biomass-hydrolyzing activity of wild and recombinant enzymes suggests that this mould is highly efficient in biomass decomposition at both moderate and high temperatures. The native enzymes produced by the mould are more efficient in activity than their mesophilic counterparts beside their low enzyme titers. The mould is able to synthesize various biomolecules, which are used in multifarious applications. Genome sequence data of M. thermophila also supported the physiological data. This review describes the biotechnological potential of thermophilic mould, M. thermophila supported by genomic and experimental evidences.

Thermophilic microorganisms in biomining.

Science.gov (United States)

Donati, Edgardo Rubén; Castro, Camila; Urbieta, María Sofía

2016-11-01

Biomining is an applied biotechnology for mineral processing and metal extraction from ores and concentrates. This alternative technology for recovering metals involves the hydrometallurgical processes known as bioleaching and biooxidation where the metal is directly solubilized or released from the matrix for further solubilization, respectively. Several commercial applications of biomining can be found around the world to recover mainly copper and gold but also other metals; most of them are operating at temperatures below 40-50 °C using mesophilic and moderate thermophilic microorganisms. Although biomining offers an economically viable and cleaner option, its share of the world´s production of metals has not grown as much as it was expected, mainly considering that due to environmental restrictions in many countries smelting and roasting technologies are being eliminated. The slow rate of biomining processes is for sure the main reason of their poor implementation. In this scenario the use of thermophiles could be advantageous because higher operational temperature would increase the rate of the process and in addition it would eliminate the energy input for cooling the system (bioleaching reactions are exothermic causing a serious temperature increase in bioreactors and inside heaps that adversely affects most of the mesophilic microorganisms) and it would decrease the passivation of mineral surfaces. In the last few years many thermophilic bacteria and archaea have been isolated, characterized, and even used for extracting metals. This paper reviews the current status of biomining using thermophiles, describes the main characteristics of thermophilic biominers and discusses the future for this biotechnology.

Alcohol dehydrogenases from thermophilic and hyperthermophilic archaea and bacteria.

Science.gov (United States)

Radianingtyas, Helia; Wright, Phillip C

2003-12-01

Many studies have been undertaken to characterise alcohol dehydrogenases (ADHs) from thermophiles and hyperthermophiles, mainly to better understand their activities and thermostability. To date, there are 20 thermophilic archaeal and 17 thermophilic bacterial strains known to have ADHs or similar enzymes, including the hypothetical proteins. Some of these thermophiles are found to have multiple ADHs, sometimes of different types. A rigid delineation of amino acid sequences amongst currently elucidated thermophilic ADHs and similar proteins is phylogenetically apparent. All are NAD(P)-dependent, with one exception that utilises the cofactor F(420) instead. Within the NAD(P)-dependent group, the thermophilic ADHs are orderly clustered as zinc-dependent ADHs, short-chain ADHs, and iron-containing/activated ADHs. Distance matrix calculations reveal that thermophilic ADHs within one type are homologous, with those derived from a single genus often showing high similarities. Elucidation of the enzyme activity and stability, coupled with structure analysis, provides excellent information to explain the relationship between them, and thermophilic ADHs diversity.

Anaerobic Thermophiles

Directory of Open Access Journals (Sweden)

Francesco Canganella

2014-02-01

Full Text Available The term “extremophile” was introduced to describe any organism capable of living and growing under extreme conditions. With the further development of studies on microbial ecology and taxonomy, a variety of “extreme” environments have been found and an increasing number of extremophiles are being described. Extremophiles have also been investigated as far as regarding the search for life on other planets and even evaluating the hypothesis that life on Earth originally came from space. The first extreme environments to be largely investigated were those characterized by elevated temperatures. The naturally “hot environments” on Earth range from solar heated surface soils and water with temperatures up to 65 °C, subterranean sites such as oil reserves and terrestrial geothermal with temperatures ranging from slightly above ambient to above 100 °C, to submarine hydrothermal systems with temperatures exceeding 300 °C. There are also human-made environments with elevated temperatures such as compost piles, slag heaps, industrial processes and water heaters. Thermophilic anaerobic microorganisms have been known for a long time, but scientists have often resisted the belief that some organisms do not only survive at high temperatures, but actually thrive under those hot conditions. They are perhaps one of the most interesting varieties of extremophilic organisms. These microorganisms can thrive at temperatures over 50 °C and, based on their optimal temperature, anaerobic thermophiles can be subdivided into three main groups: thermophiles with an optimal temperature between 50 °C and 64 °C and a maximum at 70 °C, extreme thermophiles with an optimal temperature between 65 °C and 80 °C, and finally hyperthermophiles with an optimal temperature above 80 °C and a maximum above 90 °C. The finding of novel extremely thermophilic and hyperthermophilic anaerobic bacteria in recent years, and the fact that a large fraction of them belong

Space agriculture for habitation on Mars with hyper-thermophilic aerobic composting bacteria

Science.gov (United States)

Space Agriculture Task Force; Ishikawa, Y.; Tomita-Yokotani, K.; Hashimoto, H.; Kitaya, Y.; Yamashita, M.; Nagatomo, M.; Oshima, T.; Wada, H.

Manned Mars exploration, especially for extended periods of time, will require recycle of materials to support human life. Here, a conceptual design is developed for a Martian agricultural system driven by biologically regenerative functions. One of the core biotechnologies function is the use of hyper-thermophilic aerobic composting bacterial ecology. These thermophilic bacteria can play an important role in increasing the effectiveness of the processing of human metabolic waste and inedible biomass and of converting them to fertilizer for the cultivation of plants. This microbial technology has been already well established for the purpose of processing sewage and waste materials for small local communities in Japan. One of the characteristics of the technology is that the metabolic heat release that occurs during bacterial fermentation raises the processing temperature sufficiently high at 80 100 °C to support hyper-thermophilic bacteria. Such a hyper-thermophilic system is found to have great capability of decomposing wastes including even their normally recalcitrant components, in a reasonably short period of time and of providing a better quality of fertilizer as an end-product. High quality compost has been shown to be a key element in creating a healthy regenerative food production system. In ground-based studies, the soil microbial ecology after the addition of high quality compost was shown to improve plant growth and promote a healthy symbiosis of arbuscular mycorrhizal fungi. Another advantage of such high processing temperature is the ability to sterilize the pathogenic organisms through the fermentation process and thus to secure the hygienic safety of the system. Plant cultivation is one of the other major systems. It should fully utilize solar energy received on the Martian surface for supplying energy for photosynthesis. Subsurface water and atmospheric carbon dioxide mined on Mars should be also used in the plant cultivation system. Oxygen and

In Silico Identification of Microbial Partners to Form Consortia with Anaerobic Fungi

Directory of Open Access Journals (Sweden)

St. Elmo Wilken

2018-01-01

Full Text Available Lignocellulose is an abundant and renewable resource that holds great promise for sustainable bioprocessing. However, unpretreated lignocellulose is recalcitrant to direct utilization by most microbes. Current methods to overcome this barrier include expensive pretreatment steps to liberate cellulose and hemicellulose from lignin. Anaerobic gut fungi possess complex cellulolytic machinery specifically evolved to decompose crude lignocellulose, but they are not yet genetically tractable and have not been employed in industrial bioprocesses. Here, we aim to exploit the biomass-degrading abilities of anaerobic fungi by pairing them with another organism that can convert the fermentable sugars generated from hydrolysis into bioproducts. By combining experiments measuring the amount of excess fermentable sugars released by the fungal enzymes acting on crude lignocellulose, and a novel dynamic flux balance analysis algorithm, we screened potential consortia partners by qualitative suitability. Microbial growth simulations reveal that the fungus Anaeromyces robustus is most suited to pair with either the bacterium Clostridia ljungdahlii or the methanogen Methanosarcina barkeri—both organisms also found in the rumen microbiome. By capitalizing on simulations to screen six alternative organisms, valuable experimental time is saved towards identifying stable consortium members. This approach is also readily generalizable to larger systems and allows one to rationally select partner microbes for formation of stable consortia with non-model microbes like anaerobic fungi.

Status on Science and Application of Thermophilic Anaerobic Digestion

DEFF Research Database (Denmark)

Ahring, Birgitte Kiær

1994-01-01

Thermophilic anaerobic processes are often regarded as less stable than mesophilic processes. In the paper this postulate is examined and disproved based on real operational data from of full-scale mesophilic and thermophilic biogas plants. The start-up produce for the thermophilic plants was...... for thermophilic digestion along with the implications for the methanogenic bacteria active at these temperatures....

Evaluation in Cellulolytic Activity of Stenotrophomonas sp. in Cellulose Nitrogen Free Mineral Medium

International Nuclear Information System (INIS)

Honey Thet Paing Htway; San San Yu; Zaw Ko Latt

2011-12-01

Three bacterial strains were isolated from rice rhizospheric soil and their nitrogen fixing activity was determined in nitrogen free mineral medium and broth with glucose and cellulose as carbon sources and they produced ammonium concentration (above 3ppm) in G-NFFMM and (2-3ppm) in C-NFMM. Moreover, their cellulolytic activity was determined by DNS mothod and strain H3 having the cellulolytic activity was selected. Then, cellulose, carboxymethyl cellulose, baggasse, pea haulm, corn stem, rice straw were used as substrates and determined its reducing sugar concentration. After detection of the cellulolytic activity, the bacteria produced the highest concentration of reducing sugar on cellulose substrate at 12 day incubation period with the reducing sugar amount of 0.12mg/ml and 0.298mg/ml on CMC substrates. In the study of argicultral wastes as substrates, the selected strain, H3, produced in the reducing sugar concentration with 0.12, 0.116,0.103 and 0.098mg/ml respectively. The selected strain was identified by biochemical characterists and 16s ribosomal DNA analysis and it was Stenotrophomonas sp.

Rapid establishment of thermophilic anaerobic microbial community during the one-step startup of thermophilic anaerobic digestion from a mesophilic digester.

Science.gov (United States)

Tian, Zhe; Zhang, Yu; Li, Yuyou; Chi, Yongzhi; Yang, Min

2015-02-01

The purpose of this study was to explore how fast the thermophilic anaerobic microbial community could be established during the one-step startup of thermophilic anaerobic digestion from a mesophilic digester. Stable thermophilic anaerobic digestion was achieved within 20 days from a mesophilic digester treating sewage sludge by adopting the one-step startup strategy. The succession of archaeal and bacterial populations over a period of 60 days after the temperature increment was followed by using 454-pyrosequencing and quantitative PCR. After the increase of temperature, thermophilic methanogenic community was established within 11 days, which was characterized by the fast colonization of Methanosarcina thermophila and two hydrogenotrophic methanogens (Methanothermobacter spp. and Methanoculleus spp.). At the same time, the bacterial community was dominated by Fervidobacterium, whose relative abundance rapidly increased from 0 to 28.52 % in 18 days, followed by other potential thermophilic genera, such as Clostridium, Coprothermobacter, Anaerobaculum and EM3. The above result demonstrated that the one-step startup strategy could allow the rapid establishment of the thermophilic anaerobic microbial community. Copyright © 2014 Elsevier Ltd. All rights reserved.

Drechslerella stenobrocha genome illustrates the mechanism of constricting rings and the origin of nematode predation in fungi

Science.gov (United States)

2014-01-01

Background Nematode-trapping fungi are a unique group of organisms that can capture nematodes using sophisticated trapping structures. The genome of Drechslerella stenobrocha, a constricting-ring-forming fungus, has been sequenced and reported, and provided new insights into the evolutionary origins of nematode predation in fungi, the trapping mechanisms, and the dual lifestyles of saprophagy and predation. Results The genome of the fungus Drechslerella stenobrocha, which mechanically traps nematodes using a constricting ring, was sequenced. The genome was 29.02 Mb in size and was found rare instances of transposons and repeat induced point mutations, than that of Arthrobotrys oligospora. The functional proteins involved in nematode-infection, such as chitinases, subtilisins, and adhesive proteins, underwent a significant expansion in the A. oligospora genome, while there were fewer lectin genes that mediate fungus-nematode recognition in the D. stenobrocha genome. The carbohydrate-degrading enzyme catalogs in both species were similar to those of efficient cellulolytic fungi, suggesting a saprophytic origin of nematode-trapping fungi. In D. stenobrocha, the down-regulation of saprophytic enzyme genes and the up-regulation of infection-related genes during the capture of nematodes indicated a transition between dual life strategies of saprophagy and predation. The transcriptional profiles also indicated that trap formation was related to the protein kinase C (PKC) signal pathway and regulated by Zn(2)–C6 type transcription factors. Conclusions The genome of D. stenobrocha provides support for the hypothesis that nematode trapping fungi evolved from saprophytic fungi in a high carbon and low nitrogen environment. It reveals the transition between saprophagy and predation of these fungi and also proves new insights into the mechanisms of mechanical trapping. PMID:24507587

Ras GTPases Modulate Morphogenesis, Sporulation and Cellulase Gene Expression in the Cellulolytic Fungus Trichoderma reesei

Science.gov (United States)

Zhang, Jiwei; Zhang, Yanmei; Zhong, Yaohua; Qu, Yinbo; Wang, Tianhong

2012-01-01

Background The model cellulolytic fungus Trichoderma reesei (teleomorph Hypocrea jecorina) is capable of responding to environmental cues to compete for nutrients in its natural saprophytic habitat despite its genome encodes fewer degradative enzymes. Efficient signalling pathways in perception and interpretation of environmental signals are indispensable in this process. Ras GTPases represent a kind of critical signal proteins involved in signal transduction and regulation of gene expression. In T. reesei the genome contains two Ras subfamily small GTPases TrRas1 and TrRas2 homologous to Ras1 and Ras2 from S. cerevisiae, but their functions remain unknown. Methodology/Principal Findings Here, we have investigated the roles of GTPases TrRas1 and TrRas2 during fungal morphogenesis and cellulase gene expression. We show that both TrRas1 and TrRas2 play important roles in some cellular processes such as polarized apical growth, hyphal branch formation, sporulation and cAMP level adjustment, while TrRas1 is more dominant in these processes. Strikingly, we find that TrRas2 is involved in modulation of cellulase gene expression. Deletion of TrRas2 results in considerably decreased transcription of cellulolytic genes upon growth on cellulose. Although the strain carrying a constitutively activated TrRas2G16V allele exhibits increased cellulase gene transcription, the cbh1 and cbh2 expression in this mutant still strictly depends on cellulose, indicating TrRas2 does not directly mediate the transmission of the cellulose signal. In addition, our data suggest that the effect of TrRas2 on cellulase gene is exerted through regulation of transcript abundance of cellulase transcription factors such as Xyr1, but the influence is independent of cAMP signalling pathway. Conclusions/Significance Together, these findings elucidate the functions for Ras signalling of T. reesei in cellular morphogenesis, especially in cellulase gene expression, which contribute to deciphering the

Use of Cellulolytic Marine Bacteria for Enzymatic Pretreatment in Microalgal Biogas Production

Science.gov (United States)

Muñoz, Camilo; Hidalgo, Catalina; Zapata, Manuel; Jeison, David; Riquelme, Carlos

2014-01-01

In this study, we designed and evaluated a microalgal pretreatment method using cellulolytic bacteria that naturally degrades microalgae in their native habitat. Bacterial strains were isolated from each of two mollusk species in a medium containing 1% carboxymethyl cellulose agar. We selected nine bacterial strains that had endoglucanase activity: five strains from Mytilus chilensis, a Chilean mussel, and four strains from Mesodesma donacium, a clam found in the Southern Pacific. These strains were identified phylogenetically as belonging to the genera Aeromonas, Pseudomonas, Chryseobacterium, and Raoultella. The cellulase-producing capacities of these strains were characterized, and the degradation of cell walls in Botryococcus braunii and Nannochloropsis gaditana was tested with “whole-cell” cellulolytic experiments. Aeromonas bivalvium MA2, Raoultella ornithinolytica MA5, and Aeromonas salmonicida MC25 degraded B. braunii, and R. ornithinolytica MC3 and MA5 degraded N. gaditana. In addition, N. gaditana was pretreated with R. ornithinolytica strains MC3 and MA5 and was then subjected to an anaerobic digestion process, which increased the yield of methane by 140.32% and 158.68%, respectively, over that from nonpretreated microalgae. Therefore, a “whole-cell” cellulolytic pretreatment can increase the performance and efficiency of biogas production. PMID:24795376

Thermophilic slurry-phase treatment of petroleum hydrocarbon waste sludges

International Nuclear Information System (INIS)

Castaldi, F.J.; Bombaugh, K.J.; McFarland, B.

1995-01-01

Chemoheterotrophic thermophilic bacteria were used to achieve enhanced hydrocarbon degradation during slurry-phase treatment of oily waste sludges from petroleum refinery operations. Aerobic and anaerobic bacterial cultures were examined under thermophilic conditions to assess the effects of mode of metabolism on the potential for petroleum hydrocarbon degradation. The study determined that both aerobic and anaerobic thermophilic bacteria are capable of growth on petroleum hydrocarbons. Thermophilic methanogenesis is feasible during the degradation of hydrocarbons when a strict anaerobic condition is achieved in a slurry bioreactor. Aerobic thermophilic bacteria achieved the largest apparent reduction in chemical oxygen demand, freon extractable oil, total and volatile solid,s and polycyclic aromatic hydrocarbons (PAHs) when treating oily waste sludges. The observed shift with time in the molecular weight distribution of hydrocarbon material was more pronounced under aerobic metabolic conditions than under strict anaerobic conditions. The changes in the hydrocarbon molecular weight distribution, infrared spectra, and PAH concentrations during slurry-phase treatment indicate that the aerobic thermophilic bioslurry achieved a higher degree of hydrocarbon degradation than the anaerobic thermophilic bioslurry during the same time period

Chimeric polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

Science.gov (United States)

Wogulis, Mark; Sweeney, Matthew; Heu, Tia

2017-06-14

The present invention relates to chimeric GH61 polypeptides having cellulolytic enhancing activity. The present invention also relates to polynucleotides encoding the chimeric GH61 polypeptides; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the chimeric GH61 polypeptides.

Thermotoga lettingae sp. nov. : a novel thermophilic, methanol-degrading bacterium isolated from a thermophilic anaerobic reactor

NARCIS (Netherlands)

Balk, M.; Weijma, J.; Stams, A.J.M.

2002-01-01

A novel, anaerobic, non-spore-forming, mobile, Gram-negative, thermophilic bacterium, strain TMO(T), was isolated from a thermophilic sulfate-reducing bioreactor operated at 65 degrees C with methanol as the sole substrate. The G C content of the DNA of strain TMO(T) was 39.2 molÐThe optimum pH,

Screening of highly cellulolytic fungi and the action of their cellulase enzyme systems

Energy Technology Data Exchange (ETDEWEB)

Saddler, J N

1982-11-01

Over 100 strains of wood-rotting fungi were compared for their ability to degrade wood blocks. Some of these strains were then assayed for extracellular cellulase (1,4-(1,3;1,4)-beta-D-glucan 4- glucanohydrolase, EC 3.2.1.4) activity using a variety of different solid media containing carboxymethyl cellulose or acid swollen cellulose. The diameter of clearing on these plates gave an approximate indication of the order of cellulase activities obtained from culture filtrates of these strains. Trichoderma strains grown on Vogels medium gave the highest cellulase yields. The cellulase enzyme production of T. reesei C30 and QM9414 was compared with that of eight other Trichoderma strains. Trichoderma strain E58 had comparable endoglucanase and filter paper activities with the mutant strains while the beta-D-glucosidase (beta-D-glucoside glucohydrolase, EC 3.2.1.21) activity was approximately six to nine times greater. (Refs. 26).

Thermophilic biofiltration of benzene and toluene.

Science.gov (United States)

Cho, Kyung-Suk; Yoo, Sun-Kyung; Ryu, Hee Wook

2007-12-01

In the current studies, we characterized the degradation of a hot mixture of benzene and toluene (BT) gases by a thermophilic biofilter using polyurethane as packing material and high-temperature compost as a microbial source. We also examined the effect of supplementing the biofilter with yeast extract (YE). We found that YE substantially enhanced microbial activity in the thermophilic biofilter. The degrading activity of the biofilter supplied with YE was stable during long-term operation (approximately 100 d) without accumulating excess biomass. The maximum elimination capacity (1,650 g x m(-3) h(-1)) in the biofilter supplemented with YE was 3.5 times higher than that in the biofilter without YE (470 g g x m(-3) h(-1)). At similar retention times, the capacity to eliminate BT for the YE-supplemented biofilter was higher than for previously reported mesophilic biofilters. Thus, thermophilic biofiltration can be used to degrade hydrophobic compounds such as a BT mixture. Finally, 16S rDNA polymerase chain reaction-DGGE (PCR-DGGE) fingerprinting revealed that the thermophilic bacteria in the biofilter included Rubrobacter sp. and Mycobacterium sp.

Improvement of ethanol production from crystalline cellulose via optimizing cellulase ratios in cellulolytic Saccharomyces cerevisiae.

Science.gov (United States)

Liu, Zhuo; Inokuma, Kentaro; Ho, Shih-Hsin; den Haan, Riaan; van Zyl, Willem H; Hasunuma, Tomohisa; Kondo, Akihiko

2017-06-01

Crystalline cellulose is one of the major contributors to the recalcitrance of lignocellulose to degradation, necessitating high dosages of cellulase to digest, thereby impeding the economic feasibility of cellulosic biofuels. Several recombinant cellulolytic yeast strains have been developed to reduce the cost of enzyme addition, but few of these strains are able to efficiently degrade crystalline cellulose due to their low cellulolytic activities. Here, by combining the cellulase ratio optimization with a novel screening strategy, we successfully improved the cellulolytic activity of a Saccharomyces cerevisiae strain displaying four different synergistic cellulases on the cell surface. The optimized strain exhibited an ethanol yield from Avicel of 57% of the theoretical maximum, and a 60% increase of ethanol titer from rice straw. To our knowledge, this work is the first optimization of the degradation of crystalline cellulose by tuning the cellulase ratio in a cellulase cell-surface display system. This work provides key insights in engineering the cellulase cocktail in a consolidated bioprocessing yeast strain. Biotechnol. Bioeng. 2017;114: 1201-1207. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Interactions of fungi from fermented sausage with regenerated cellulose casings.

Science.gov (United States)

Sreenath, Hassan K; Jeffries, Thomas W

2011-11-01

This research examined cellulolytic effects of fungi and other microbes present in cured sausages on the strength and stability of regenerated cellulose casings (RCC) used in the sausage industry. Occasionally during the curing process, RCC would split or fail, thereby leading to loss of product. The fungus Penicillium sp. BT-F-1, which was isolated from fermented sausages, and other fungi, which were introduced to enable the curing process, produced small amounts of cellulases on RCC in both liquid and solid cultivations. During continued incubation for 15-60 days in solid substrate cultivation (SSC) on RCC support, the fungus Penicillium sp isolate BT-F-1 degraded the casings' dry weights by 15-50% and decreased their tensile strengths by ~75%. Similarly commercial cellulase(s) resulted in 20-50% degradation of RCC in 48 h. During incubation with Penicillium sp BT-F-1, the surface structure of RCC collapsed, resulting in loss of strength and stability of casings. The matrix of industrial RCC comprised 88-93% glucose polymer residues with 0.8-4% xylan impurities. Premature casing failure appeared to result from operating conditions in the manufacturing process that allowed xylan to build up in the extrusion bath. The sausage fungus Penicillium sp BT-F-1 produced xylanases to break down soft xylan pockets prior to slow cellulosic dissolution of RCC.

Survival of thermophilic and hyper-thermophilic microorganisms after exposure to UV-C, ionizing radiation and desiccation

International Nuclear Information System (INIS)

Beblo, K.; Wirth, R.; Huber, H.; Douki, T.; Schmalz, G.; Rachel, R.

2011-01-01

In this study, we investigated the ability of several (hyper-) thermophilic Archaea and phylo-genetically deep-branching thermophilic Bacteria to survive high fluences of monochromatic UV-C (254 nm) and high doses of ionizing radiation, respectively. Nine out of fourteen tested microorganisms showed a surprisingly high tolerance against ionizing radiation, and two species (Aquifex pyrophilus and Ignicoccus hospitalis) were even able to survive 20 kGy. Therefore, these species had a comparable survivability after exposure to ionizing radiation such as Deinococcus radiodurans. In contrast, there was nearly no difference in survival of the tested strains after exposure to UV-C under anoxic conditions. If the cells had been dried in advance of UV-C irradiation, they were more sensitive to UV-C radiation compared with cells irradiated in liquid suspension; this effect could be reversed by the addition of protective material like sulfidic ores before irradiation. By exposure to UV-C, photoproducts were formed in the DNA of irradiated Archaea and Bacteria. The distribution of the main photoproducts was species specific, but the amount of the photoproducts was only partly dependent on the applied fluence. Overall, our results show that tolerance to radiation seems to be a common phenomenon among thermophilic and hyper-thermophilic microorganisms. (authors)

Thermophilic Sulfate-Reducing Bacteria in Cold Marine Sediment

DEFF Research Database (Denmark)

ISAKSEN, MF; BAK, F.; JØRGENSEN, BB

1994-01-01

sulfate-reducing bacteria was detected. Time course experiments showed constant sulfate reduction rates at 4 degrees C and 30 degrees C, whereas the activity at 60 degrees C increased exponentially after a lag period of one day. Thermophilic, endospore-forming sulfate-reducing bacteria, designated strain...... C to search for presence of psychrophilic, mesophilic and thermophilic sulfate-reducing bacteria. Detectable activity was initially only in the mesophilic range, but after a lag phase sulfate reduction by thermophilic sulfate-reducing bacteria were observed. No distinct activity of psychrophilic...... P60, were isolated and characterized as Desulfotomaculum kuznetsovii. The temperature response of growth and respiration of strain P60 agreed well with the measured sulfate reduction at 50 degrees-70 degrees C. Bacteria similar to strain P60 could thus be responsible for the measured thermophilic...

Thermophilic Sulfate-Reducing Bacteria in Cold Marine Sediment

DEFF Research Database (Denmark)

ISAKSEN, MF; BAK, F.; JØRGENSEN, BB

1994-01-01

C to search for presence of psychrophilic, mesophilic and thermophilic sulfate-reducing bacteria. Detectable activity was initially only in the mesophilic range, but after a lag phase sulfate reduction by thermophilic sulfate-reducing bacteria were observed. No distinct activity of psychrophilic...... sulfate-reducing bacteria was detected. Time course experiments showed constant sulfate reduction rates at 4 degrees C and 30 degrees C, whereas the activity at 60 degrees C increased exponentially after a lag period of one day. Thermophilic, endospore-forming sulfate-reducing bacteria, designated strain...... P60, were isolated and characterized as Desulfotomaculum kuznetsovii. The temperature response of growth and respiration of strain P60 agreed well with the measured sulfate reduction at 50 degrees-70 degrees C. Bacteria similar to strain P60 could thus be responsible for the measured thermophilic...

Utilization of steam- and explosion-decompressed aspen wood by some anaerobes. [Acetivibrio cellulolyticus, Clostridium saccharolyticum, Zymomonas anaerobia

Energy Technology Data Exchange (ETDEWEB)

Khan, A W; Asther, M; Giuliano, C

1984-01-01

Tests made to study the suitability of using steam- and explosion-decompressed aspen wood as substrate in anaerobic fermentations indicated that after washing with dilute NaOH it becomes over 80% accessible to both mesophilic and thermophilic cellulolytic anaerobes and cellulases, compared with delignified, ball-milled pulp. After washing, this material was also found to be suitable for the single-step conversion of cellulose to ethanol using cocultures consisting of cellylolytic and ethanol-producing saccharolytic anaerobes; and without and after washing by the use of cellulolytic enzymes and ethanologenic anaerobes. 16 references, 3 tables.

Degradation of cellulosic substances by Thermomonospora curvata

Energy Technology Data Exchange (ETDEWEB)

Stutzenberger, F J

1979-05-01

Research is reported on the cellulolytic activity of Thermomonospora curvata, a thermophilic cellulolytic actinomycete prevalent in municipal solid waste compost. Various cellulosic wastes were evaluated for their potential for the induction of cellulase synthesis by Th. curvata and the extent of cellulose degradation under optimal culture conditions. All the substrates tested showed significant degradation of their cellulose content with the exception of sawdust and barley straw. In contrast to Trichoderma viride, cotton fibers were the best substrates for both C/sub 1/ and C/sub x/ cellulase production. Further research is recommended. (JSR)

The mutagenic action of UV-light irradiation on aspergillus terreus in relation to antibacterial activity

International Nuclear Information System (INIS)

Ouda, S.M.

2009-01-01

Four strains of cellulolytic fungi (i.e Penicillium oxalicum, Aspergillus niger, Aspergillus terreus and Trichoderma longibrachiatum) were tested for the production of cellulolytic enzymes and antibiotic action. these fungi were cultured on Czapek Dox's media with different cellulosic substrates. A. terreus. exhibited the highest cellulolytic activity and the highest level of anti-bacterial activity against Staphylococcus aureus. and Escherichia Coli, Ultraviolet light as a mutagenic treatment for A.terreus was investigated. Two treated isolates U.30.12 and U.60.10 were obtained after the treatment at dose 30 and 60 min., respectively with a higher antibiotic activity in comparison with the wild isolate. A compound of fifteen carbon atom of terrecylic acid was isolated from ethyl acetate extract using spectroscopic analysis

One-step production of biocommodities from lignocellulosic biomass by recombinant cellulolytic bacillus subtilis: opportunities and challenges

Energy Technology Data Exchange (ETDEWEB)

Zhang, Xiao-Zhou [Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA (United States); Zhang, Yi-Heng P. [Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA (United States); Institute for Critical Technology and Applied Science, Virginia Polytechnic Institute and State University, Blacksburg, VA (United States); BioEnergy Science Center of Department of Energy, Oak Ridge, TN (United States)

2010-10-15

One-step consolidated bioprocessing that integrates cellulase production, cellulose hydrolysis, and product fermentation into a single step for decreasing costly cellulase use, increasing volumetric productivity, and reducing capital investment is widely accepted for low-cost production of biofuels or other value-added biochemicals. Considering the narrow margins between biomass and low-value biocommodities, good physiological performance of industrial microbes is crucial for economically viable production. Bacillus subtilis, the best-characterized Gram-positive microorganism, is a major industrial microorganism with numerous valuable features such as hexose and pentose utilization, low-nutrient needs, fast growth rate, high protein secretion capacity, industrial safety, etc. As compared with other potential consolidated bioprocessing microorganisms such as Clostridium spp., Escherichia coli, and the yeast Saccharomyces cerevisiae, recombinant cellulolytic B. subtilis strains would be a potential platform for biocommodity production from nonfood biomass. Here, we review the advances in recombinant cellulolytic B. subtilis development and metabolic engineering for biocommodity production, and discuss the opportunities and challenges of cellulolytic B. subtilis for biocommodity production. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

Cloning and recombinant expression of a cellulase from the cellulolytic strain Streptomyces sp. G12 isolated from compost

Directory of Open Access Journals (Sweden)

Amore Antonella

2012-12-01

Full Text Available Abstract Background The use of lignocellulosic materials for second generation ethanol production would give several advantages such as minimizing the conflict between land use for food and fuel production, providing less expensive raw materials than conventional agricultural feedstock, allowing lower greenhouse gas emissions than those of first generation ethanol. However, cellulosic biofuels are not produced at a competitive level yet, mainly because of the high production costs of the cellulolytic enzymes. Therefore, this study was aimed at discovering new cellulolytic microorganisms and enzymes. Results Different bacteria isolated from raw composting materials obtained from vegetable processing industry wastes were screened for their cellulolytic activity on solid medium containing carboxymethylcellulose. Four strains belonging to the actinomycetes group were selected on the basis of their phenotypic traits and cellulolytic activity on solid medium containing carboxymethylcellulose. The strain showing the highest cellulolytic activity was identified by 16S rRNA sequencing as belonging to Streptomyces genus and it was designated as Streptomyces sp. strain G12. Investigating the enzymes responsible for cellulase activity produced by Streptomyces G12 by proteomic analyses, two endoglucanases were identified. Gene coding for one of these enzymes, named CelStrep, was cloned and sequenced. Molecular analysis showed that the celstrep gene has an open reading frame encoding a protein of 379 amino acid residues, including a signal peptide of 37 amino acid residues. Comparison of deduced aminoacidic sequence to the other cellulases indicated that the enzyme CelStrep can be classified as a family 12 glycoside hydrolase. Heterologous recombinant expression of CelStrep was carried out in Escherichia coli, and the active recombinant enzyme was purified from culture supernatant and characterized. It catalyzes the hydrolysis of carboxymethylcellulose

Cloning and recombinant expression of a cellulase from the cellulolytic strain Streptomyces sp. G12 isolated from compost

Science.gov (United States)

2012-01-01

Background The use of lignocellulosic materials for second generation ethanol production would give several advantages such as minimizing the conflict between land use for food and fuel production, providing less expensive raw materials than conventional agricultural feedstock, allowing lower greenhouse gas emissions than those of first generation ethanol. However, cellulosic biofuels are not produced at a competitive level yet, mainly because of the high production costs of the cellulolytic enzymes. Therefore, this study was aimed at discovering new cellulolytic microorganisms and enzymes. Results Different bacteria isolated from raw composting materials obtained from vegetable processing industry wastes were screened for their cellulolytic activity on solid medium containing carboxymethylcellulose. Four strains belonging to the actinomycetes group were selected on the basis of their phenotypic traits and cellulolytic activity on solid medium containing carboxymethylcellulose. The strain showing the highest cellulolytic activity was identified by 16S rRNA sequencing as belonging to Streptomyces genus and it was designated as Streptomyces sp. strain G12. Investigating the enzymes responsible for cellulase activity produced by Streptomyces G12 by proteomic analyses, two endoglucanases were identified. Gene coding for one of these enzymes, named CelStrep, was cloned and sequenced. Molecular analysis showed that the celstrep gene has an open reading frame encoding a protein of 379 amino acid residues, including a signal peptide of 37 amino acid residues. Comparison of deduced aminoacidic sequence to the other cellulases indicated that the enzyme CelStrep can be classified as a family 12 glycoside hydrolase. Heterologous recombinant expression of CelStrep was carried out in Escherichia coli, and the active recombinant enzyme was purified from culture supernatant and characterized. It catalyzes the hydrolysis of carboxymethylcellulose following a Michaelis�

Energy transduction and transport processes in thermophilic bacteria

NARCIS (Netherlands)

Konings, W. N.; Tolner, B.; Speelmans, G.; Elferink, M. G. L.; de Wit, J. G.; Driessen, A. J. M.

1992-01-01

Bacterial growth at the extremes of temperature has remained a fascinating aspect in the study of membrane function and structure. The stability of the integral membrane proteins of thermophiles make them particularly amenable to study. Respiratory enzymes of thermophiles appear to be functionally

Interactions between Cellulolytic Enzymes with Native, Autohydrolysis, and Technical Lignins and the Effect of a Polysorbate Amphiphile in Reducing Nonproductive Binding.

Science.gov (United States)

Fritz, Consuelo; Ferrer, Ana; Salas, Carlos; Jameel, Hasan; Rojas, Orlando J

2015-12-14

Understanding enzyme-substrate interactions is critical in designing strategies for bioconversion of lignocellulosic biomass. In this study we monitored molecular events, in situ and in real time, including the adsorption and desorption of cellulolytic enzymes on lignins and cellulose, by using quartz crystal microgravimetry and surface plasmon resonance. The effect of a nonionic surface active molecule was also elucidated. Three lignin substrates relevant to the sugar platform in biorefinery efforts were considered, namely, hardwood autohydrolysis cellulolytic (HWAH), hardwood native cellulolytic (MPCEL), and nonwood native cellulolytic (WSCEL) lignin. In addition, Kraft lignins derived from softwoods (SWK) and hardwoods (HWK) were used as references. The results indicated a high affinity between the lignins with both, monocomponent and multicomponent enzymes. More importantly, the addition of nonionic surfactants at concentrations above their critical micelle concentration reduced remarkably (by over 90%) the nonproductive interactions between the cellulolytic enzymes and the lignins. This effect was hypothesized to be a consequence of the balance of hydrophobic and hydrogen bonding interactions. Moreover, the reduction of surface roughness and increased wettability of lignin surfaces upon surfactant treatment contributed to a lower affinity with the enzymes. Conformational changes of cellulases were observed upon their adsorption on lignin carrying preadsorbed surfactant. Weak electrostatic interactions were determined in aqueous media at pH between 4.8 and 5.5 for the native cellulolytic lignins (MPCEL and WSCEL), whereby a ∼20% reduction in the enzyme affinity was observed. This was mainly explained by electrostatic interactions (osmotic pressure effects) between charged lignins and cellulases. Noteworthy, adsorption of nonionic surfactants onto cellulose, in the form cellulose nanofibrils, did not affect its hydrolytic conversion. Overall, our results

Presence and transcriptional activity of anaerobic fungi in agricultural biogas plants.

Science.gov (United States)

Dollhofer, Veronika; Callaghan, Tony M; Griffith, Gareth W; Lebuhn, Michael; Bauer, Johann

2017-07-01

Bioaugmentation with anaerobic fungi (AF) is promising for improved biogas generation from lignocelluloses-rich substrates. However, before implementing AF into biogas processes it is necessary to investigate their natural occurrence, community structure and transcriptional activity in agricultural biogas plants. Thus, AF were detected with three specific PCR based methods: (i) Copies of their 18S genes were found in 7 of 10 biogas plants. (ii) Transcripts of a GH5 endoglucanase gene were present at low level in two digesters, indicating transcriptional cellulolytic activity of AF. (iii) Phylogeny of the AF-community was inferred with the 28S gene. A new Piromyces species was isolated from a PCR-positive digester. Evidence for AF was only found in biogas plants operated with high proportions of animal feces. Thus, AF were most likely transferred into digesters with animal derived substrates. Additionally, high process temperatures in combination with long retention times seemed to impede AF survival and activity. Copyright © 2017 Elsevier Ltd. All rights reserved.

Turning cellulose waste into electricity: hydrogen conversion by a hydrogenase electrode.

Directory of Open Access Journals (Sweden)

Sergey M Abramov

Full Text Available Hydrogen-producing thermophilic cellulolytic microorganisms were isolated from cow faeces. Rates of cellulose hydrolysis and hydrogen formation were 0.2 mM L(-1 h(-1 and 1 mM L(-1 h(-1, respectively. An enzymatic fuel cell (EFC with a hydrogenase anode was used to oxidise hydrogen produced in a microbial bioreactor. The hydrogenase electrode was exposed for 38 days (912 h to a thermophilic fermentation medium. The hydrogenase activity remaining after continuous operation under load was 73% of the initial value.

Turning Cellulose Waste Into Electricity: Hydrogen Conversion by a Hydrogenase Electrode

Science.gov (United States)

Abramov, Sergey M.; Sadraddinova, Elmira R.; Shestakov, Andrey I.; Voronin, Oleg G.; Karyakin, Arkadiy A.; Zorin, Nikolay A.; Netrusov, Alexander I.

2013-01-01

Hydrogen-producing thermophilic cellulolytic microorganisms were isolated from cow faeces. Rates of cellulose hydrolysis and hydrogen formation were 0.2 mM L-1 h-1 and 1 mM L-1 h-1, respectively. An enzymatic fuel cell (EFC) with a hydrogenase anode was used to oxidise hydrogen produced in a microbial bioreactor. The hydrogenase electrode was exposed for 38 days (912 h) to a thermophilic fermentation medium. The hydrogenase activity remaining after continuous operation under load was 73% of the initial value. PMID:24312437

The improvement of eggs quality of Mojosari duck (Anas javanica with soybean husk fermentation using cellulolytic bacteria of Spodoptera litura

Directory of Open Access Journals (Sweden)

Sri Hidanah

2018-05-01

Full Text Available Aim: This study was aimed to improve the quality of the eggs of Mojosari duck (Anas javanica through complete feeding containing soybean husk was fermented using cellulolytic bacteria of Spodoptera litura. Materials and Methods: This study consisted of three stages: The first stages, isolation and identification of cellulolytic bacteria from S. litura; the second stage, the fermentation of soybean husk through the application of bacterial cellulolytic isolate from the first stage; and the third stage, the application of the best complete feed formulation from the second stage to Mojosari duck. Results: There are four dominant bacteria: Bacillus sp., Cellulomonas sp., Pseudomonas sp., and Cytophaga sp. Furthermore, the best reduction of the crude fiber of soybean husks is the use of Cellulomonas sp. bacteria. The final of the study, the quality of the eggs of Anas javanica, was improved, as indicated by cholesterol decrease from the yolk without the decrease of egg weight and eggshell thickness, although the decrease in egg yolk color was inevitable. Conclusion: Soy husk fermentation using cellulolytic bacteria of S. litura was added to complete feeding can be performed to improve the quality of the eggs of Mojosari duck.

Mesophilic and thermophilic anaerobic digestion of sulphate-containing wastewaters.

Science.gov (United States)

Colleran, E; Pender, S

2002-01-01

The effect of sulphate at an influent chemical oxygen demand (COD):sulphate ratio of 4 on the operational performance of anaerobic hybrid reactors treating molasses wastewater was investigated under mesophilic and thermophilic conditions in a long-term laboratory-scale study over a 1,081 day period. The presence of sulphate reduced the COD removal efficiency under both mesophilic and thermophilic conditions. At 55 degrees C, effluent acetate levels were consistently greater than 4000 mg l(-1) indicating that thermophilic acetate-utilising methane-producing bacteria (MPB) or sulphate-reducing bacteria (SRB) had not developed in the reactor under the conditions applied. At 37 degrees C, acetate was exclusively utilised by acetoclastic methanogens, whereas H2-utilising SRB predominated over H2-utilising MPB in the competition for hydrogen. By contrast, hydrogenotrophic MPB were shown to outcompete H2-utilising SRB during long-term thermophilic operation. 16SrDNA analysis of the seed sludge and reactor biomass on conclusion of the 37 degrees C and 55 degrees C trials illustrated that the dominant methanogen present on conclusion of the thermophilic trial in the absence of influent sulphate was related to Methanocorpusculum parvuum, and was capable of growth on both acetate and hydrogen. By contrast, an organism closely related to Methanobacterium thermoautotrophicum was the dominant methanogen present in the sulphate-fed reactor on completion of the thermophilic trial.

Comparative economic assessment of ethanol production under mesophilic and thermophilic conditions

International Nuclear Information System (INIS)

Mistry, P.B.

1991-01-01

Key technical factors affecting the economics of bioethanol production are critically analyzed with special reference to the relative merits of thermophilic and mesophilic fermentation. A number of novel process schemes to take advantage of thermophilic operation are discussed. Analysis of the capital and operating costs for a range of flowsheets then provides a basis for critical study. Estimates for thermophilic production are compared with those for a sugar cane based mesophilic process (using S. cerevisiae). For the thermophilic fermentation, the basic kinetic and yield constants are based on projected values for a strain of B. stearothermophilus. Compared to mesophilic operation, thermophilic operation results in reduced capital, operating and feed costs. The feed cost still accounts for a large proportion (75%) of the total production cost. However, on a feed-cost-free basis, a reduction in production cost of up to 32% could be realized by changing to thermophilic operation from existing yeast-based processes, after minor process modifications. 20 refs., 10 figs., 8 tabs

Thermophilic (55 - 65°C) and extreme thermophilic (70 - 80°C) sulfate reduction in methanol and formate-fed UASB reactors

NARCIS (Netherlands)

Vallero, M.V.G.; Camarero, E.; Lettinga, G.; Lens, P.N.L.

2004-01-01

The feasibility of thermophilic (55-65 degreesC) and extreme thermophilic (70-80 degreesC) sulfate-reducing processes was investigated in three lab-scale upflow anaerobic sludge bed (UASB) reactors fed with either methanol or formate as the sole substrates and inoculated with mesophilic granular

Isolation of aerobic cultivable cellulolytic bacteria from different regions of the gastrointestinal tract of giant land snail Achatina fulica

OpenAIRE

Guilherme L. Pinheiro; Guilherme L. Pinheiro; Raquel eCorrea; Raquel eSoares; Alexander eCardoso; Catia eChaia; Mayssa M Clementino; Eloi eGarcia; Wanderley eDe Souza; Wanderley eDe Souza; Susana eFrases; Susana eFrases

2015-01-01

The enzymatic hydrolysis of cellulose by cellulases is one of the major limiting steps in the conversion of lignocellulosic biomass to yield bioethanol. To overcome this hindrance, significant efforts are underway to identify novel cellulases. The snail Achatina fulica is a gastropod with high cellulolytic activity, mainly due to the abundance of glycoside hydrolases produced by both the animal and its resident microbiota. In this study, we partially assessed the cellulolytic bacterial divers...

Vertical zonation and seed germination indices of chromium resistant cellulolytic and nitrogen fixing bacteria from a chronically metal exposed land area

International Nuclear Information System (INIS)

Aslam, S.; Qazi, J.I.

2014-01-01

Twenty eight cellulolytic and 25 nitrogen fixing bacteria were isolated from 20, 40 and 60 cm depths of the chromium contaminated land area. The cellulolytic as well as nitrogen fixing microbial communities in soil profiles were dominated by genus Bacillus. More diverse nitrogen fixing bacterial isolates belonging to different genera Paenibacillus, Corynebacterium and Pseudomonas were observed as compared to cellulolytic bacterial community. Majority of the cellulolytic bacteria were found inhabitants of 20 cm soil layer while 40 cm depth was the preferred zone for the nitrogen fixing bacteria. Screening of the bacterial isolates for chromium resistance showed that isolates designated as ASK15 and ASK16 were able to resist up to 1800 mg/l of chromium while the nitrogen fixing isolates which offered a maximum resistant level up to 1650 mg/l of chromium were ASNt10 and ASNS13. Nitrogen fixing isolates enhanced seed germination by 33% and expressed efficient nitrogenase activity up to 0.80 (C/sub 2/H/sub 2/ nmol/ml/hr). Growth promoting assay proved ASNt10 a potential isolate which produced 90 meu g/ml of indoleacetic acid (IAA). Though cellulolytic isolates did not affect seed germination, a significant influence on root length similar to that of ASNt10 and ASNS13 with nearly 5-fold increase in comparison with uninoculated control was observed. The isolates ASK15, ASK16 were identified as Bacillus cereus while ASNt10 and ASNS13 as Paenibacillus barcinonensis and Bacillus megaterium, respectively. (author)

Coculture fermentation of banana agro-waste to ethanol by ...

African Journals Online (AJOL)

Banana is a major cash crop of many regions generating good amount of waste after harvest. This agro waste which is left for natural degradation is used as substrate for single step ethanol fermentation by thermophilic, cellulolytic, ethanologenic Clostridium thermocellum CT2, a new culture isolated from elephant ...

Improving biogas production from anaerobic co-digestion of Thickened Waste Activated Sludge (TWAS) and fat, oil and grease (FOG) using a dual-stage hyper-thermophilic/thermophilic semi-continuous reactor.

Science.gov (United States)

Alqaralleh, Rania Mona; Kennedy, Kevin; Delatolla, Robert

2018-07-01

This paper investigates the feasibility and advantages of using a dual-stage hyper-thermophilic/thermophilic semi-continuous reactor system for the co-digestion of Thickened Waste Activated Sludge (TWAS) and Fat, Oil and Grease (FOG) to produce biogas in high quantity and quality. The performance of the dual-stage hyper-thermophilic (70°C)/thermophilic (55°C) anaerobic co-digestion system is evaluated and compared to the performance of a single-stage thermophilic (55°C) reactor that was used to co-digest the same FOG-TWAS mixtures. Both co-digestion reactors were compared to a control reactor (the control reactor was a single-stage thermophilic reactor that only digested TWAS). The effect of FOG% in the co-digestion mixture (based on total volatile solids) and the reactor hydraulic retention time (HRT) on the biogas/methane production and the reactors' performance were thoroughly investigated. The FOG% that led to the maximum methane yield with a stable reactor performance was determined for both reactors. The maximum FOG% obtained for the single-stage thermophilic reactor at 15 days HRT was found to be 65%. This 65% FOG resulted in 88.3% higher methane yield compared to the control reactor. However, the dual-stage hyper-thermophilic/thermophilic co-digestion reactor proved to be more efficient than the single-stage thermophilic co-digestion reactor, as it was able to digest up to 70% FOG with a stable reactor performance. The 70% FOG in the co-digestion mixture resulted in 148.2% higher methane yield compared to the control at 15 days HRT. 70% FOG (based on total volatile solids) is so far the highest FOG% that has been proved to be useful and safe for semi-continuous reactor application in the open literature. Finally, the dual-stage hyper-thermophilic/thermophilic co-digestion reactor also proved to be efficient and stable in co-digesting 40% FOG mixtures at lower HRTs (i.e., 9 and 12 days) and still produce high methane yields and Class A effluents

Production of cellulolytic enzymes from ascomycetes

DEFF Research Database (Denmark)

Hansen, Gustav Hammerich; Lübeck, Mette; Frisvad, Jens Christian

2015-01-01

Optimizing production of cellulose degrading enzymes is of great interest in order to increase the feasibility of constructing biorefinery facilities for a sustainable supply of energy and chemical products. The ascomycete phylum has a large potential for the production of cellulolytic enzymes....... Although numerous enzymatic profiles have already been unraveled, the research has been covering only a limited number of species and genera, thus leaving many ascomycetes to be analyzed. Such analysis requires choosing appropriate media and cultivation methods that ensure enzyme profiles with high...... specificities and activities. However, the choice of media, cultivation methods and enzyme assays highly affect the enzyme activity profile observed. This review provides an overview of enzymatic profiles for several ascomycetes covering phylogenetically distinct genera and species. The profiles of cellulose...

Clostridium thermocellum: adhesion and sporulation while adhered to cellulose and hemicellulose

Energy Technology Data Exchange (ETDEWEB)

Wiegel, J.; Dykstra, M.

1984-01-01

During growth in the presence of fibers composed of cellulose or hemicellulose, various strains of the thermophilic soil bacterium Clostridium thermocellum and several newly isolated thermophilic anaerobic soil bacteria adhered to the fibers. Attachment occurred via a fibrous ruthenium red-staining material. C. thermocellum sporulated while attached to the fibers when the pH dropped below 6.4. It is postulated that the attachment is involved in cellulose breakdown and that C. thermocellum gaines an advantage by remaining attached to its insoluble substrates when the environment is not suitable for rapid growth. The tendency to adhere to cellulose fibers was used in the purification of thermophilic cellulolytic anaerobes. 27 references, 7 figures.

Thermophillic Sidestream Anaerobic Membrane Bioreactors: The Shear Rate Dilemma

NARCIS (Netherlands)

Jeison, D.A.; Telkamp, P.; Lier, van J.B.

2009-01-01

Anaerobic biomass retention under thermophilic conditions has proven difficult. Membrane filtration can be used as alternative way to achieve high sludge concentrations. This research studied the feasibility of anaerobic membrane bioreactors (AnMBRs) under thermophilic conditions. A sidestream MBR

Hydrophobic environment is a key factor for the stability of thermophilic proteins.

Science.gov (United States)

Gromiha, M Michael; Pathak, Manish C; Saraboji, Kadhirvel; Ortlund, Eric A; Gaucher, Eric A

2013-04-01

The stability of thermophilic proteins has been viewed from different perspectives and there is yet no unified principle to understand this stability. It would be valuable to reveal the most important interactions for designing thermostable proteins for such applications as industrial protein engineering. In this work, we have systematically analyzed the importance of various interactions by computing different parameters such as surrounding hydrophobicity, inter-residue interactions, ion-pairs and hydrogen bonds. The importance of each interaction has been determined by its predicted relative contribution in thermophiles versus the same contribution in mesophilic homologues based on a dataset of 373 protein families. We predict that hydrophobic environment is the major factor for the stability of thermophilic proteins and found that 80% of thermophilic proteins analyzed showed higher hydrophobicity than their mesophilic counterparts. Ion pairs, hydrogen bonds, and interaction energy are also important and favored in 68%, 50%, and 62% of thermophilic proteins, respectively. Interestingly, thermophilic proteins with decreased hydrophobic environments display a greater number of hydrogen bonds and/or ion pairs. The systematic elimination of mesophilic proteins based on surrounding hydrophobicity, interaction energy, and ion pairs/hydrogen bonds, led to correctly identifying 95% of the thermophilic proteins in our analyses. Our analysis was also applied to another, more refined set of 102 thermophilic-mesophilic pairs, which again identified hydrophobicity as a dominant property in 71% of the thermophilic proteins. Further, the notion of surrounding hydrophobicity, which characterizes the hydrophobic behavior of residues in a protein environment, has been applied to the three-dimensional structures of elongation factor-Tu proteins and we found that the thermophilic proteins are enriched with a hydrophobic environment. The results obtained in this work highlight the

Assessment of cellulolytic microorganisms in soils of Nevados Park, Colombia

OpenAIRE

Avellaneda-Torres,Lizeth Manuela; Pulido,Claudia Patricia Guevara; Rojas,Esperanza Torres

2014-01-01

A systematized survey was conducted to find soil-borne microbes that degrade cellulose in soils from unique ecosystems, such as the Superpáramo, Páramo, and the High Andean Forest in the Nevados National Natural Park (NNNP), Colombia. These high mountain ecosystems represent extreme environments, such as high levels of solar radiation, low atmospheric pressure, and extreme daily changes in temperature. Cellulolytic activity of the microorganisms was evaluated using qualitative tests, such as ...

Bioprospecting thermophiles for cellulase production: a review.

Science.gov (United States)

Acharya, Somen; Chaudhary, Anita

2012-07-01

Most of the potential bioprospecting is currently related to the study of the extremophiles and their potential use in industrial processes. Recently microbial cellulases find applications in various industries and constitute a major group of industrial enzymes. Considerable amount of work has been done on microbial cellulases, especially with resurgence of interest in biomass ethanol production employing cellulases and use of cellulases in textile and paper industry. Most efficient method of lignocellulosic biomass hydrolysis is through enzymatic saccharification using cellulases. Significant information has also been gained about the physiology of thermophilic cellulases producers and process development for enzyme production and biomass saccharification. The review discusses the current knowledge on cellulase producing thermophilic microorganisms, their physiological adaptations and control of cellulase gene expression. It discusses the industrial applications of thermophilic cellulases, their cost of production and challenges in cellulase research especially in the area of improving process economics of enzyme production.

Relationship between microbial community dynamics and process performance during thermophilic sludge bioleaching.

Science.gov (United States)

Chen, Shen-Yi; Chou, Li-Chieh

2016-08-01

Heavy metals can be removed from the sludge using bioleaching technologies at thermophilic condition, thereby providing an option for biotreatment of wasted sludge generated from wastewater treatment. The purposes of this study were to establish a molecular biology technique, real-time PCR, for the detection and enumeration of the sulfur-oxidizing bacteria during the thermophilic sludge bioleaching. The 16S rRNA gene for real-time PCR quantification targeted the bioleaching bacteria: Sulfobacillus thermosulfidooxidans, Sulfobacillus acidophilus, and Acidithiobacillus caldus. The specificity and stringency for thermophilic sulfur-oxidizing bacteria were tested before the experiments of monitoring the bacterial community, bacterial number during the thermophilic sludge bioleaching and the future application on testing various environmental samples. The results showed that S. acidophilus was identified as the dominant sulfur-oxidizing bacteria, while A. caldus and S. thermosulfidooxidans occurred in relatively low numbers. The total number of the sulfur-oxidizing bacteria increased during the thermophilic bioleaching process. Meanwhile, the decrease of pH, production of sulfate, degradation of SS/VSS, and solubilization of heavy metal were found to correlate well with the population of thermophilic sulfur-oxidizing bacteria during the bioleaching process. The real-time PCR used in this study is a suitable method to monitor numbers of thermophilic sulfur-oxidizing bacteria during the bioleaching process.

Comparing Residue Clusters from Thermophilic and Mesophilic Enzymes Reveals Adaptive Mechanisms.

Science.gov (United States)

Sammond, Deanne W; Kastelowitz, Noah; Himmel, Michael E; Yin, Hang; Crowley, Michael F; Bomble, Yannick J

2016-01-01

Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research. Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. Thus the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions.

State of the art and future perspectives of thermophilic anaerobic digestion

DEFF Research Database (Denmark)

Ahring, Birgitte Kiær; Mladenovska, Zuzana; Iranpour, R.

2002-01-01

The slate of the art of thermophilic digestion is discussed. Thermophilic digestion is a well established technology in Europe for treatment of mixtures of waste in common large scale biogas plants or for treatment of the organic fraction of municipal solid waste. Due to a large number of failures...... over time with thermophilic digestion of sewage sludge this process has lost its appeal in the USA. New demands on sanitation of biosolids before land use will, however, bring the attention back to the use of elevated temperatures during sludge stabilization. In the paper we show how the use of a start......-up strategy based on the actual activity of key microbes can be used to ensure proper and fast transfer of mesophilic digesters into thermophilic operation. Extreme thermophilic temperatures of 65degreesC or more may be necessary in the future to meet the demands for full sanitation of the waste material...

Relationship between soil cellulolytic activity and suppression of seedling blight of barley in arable soils

DEFF Research Database (Denmark)

Rasmussen, Peter Have; Knudsen, I.; Elmholt, S.

2002-01-01

the Hanes-Wolf transformation of the Michaelis-Menten equation. Soil samples from 6 to 13 cm depth were collected in the early spring as undisturbed blocks from 10 arable soils with different physico-chemical properties and cultivation history. Significant correlations were found between soil suppresiveness......The objective was to investigate the relationship between soil suppression of seedling blight of barley caused by Fusarium culmorum (W.G. Smith) Sacc. and the soil cellulolytic activity of beta-glucosidase, cellobiohydrolase and endocellulase. Disease suppression was investigated in bioassays...... with test soils mixed with sand, and barley seeds inoculated with F. culmorum. After 19 days, disease severity was evaluated on the barley seedlings. Soil cellulolytic activities were measured using 4-methylumbelliferyl-labelled fluorogenic substrates, and were expressed as V-max values obtained by using...

Cellulolytic and Butyrivibrio fibrisolvens bacteria population density, after supplementing fodder diets (Pennisetum clandestinum

Directory of Open Access Journals (Sweden)

Licet Molina G

2015-11-01

Full Text Available Objective. Determine the population density of cellulolytic bacteria, Butyrivibrio fibrisolvens and the concentration of vaccenic acid, by supplementing diets consisting of kikuyu grass (Pennisetum clandestinum Hoechst. Ex Chiov. as base ingredient, together with cassava flour and biomass (effluent from ethanol production in rumen simulator-Rusitec. Materials and methods. Four treatments (T were evaluated, these were composed as: T1/Control 1: 100% kikuyu grass with a total protein intake of 23.9%, T2: a mixture of 70% kikuyu grass, 20% biomass and 10% cassava flour with a total protein intake of 19.4%; T3/Control 2: 100% kikuyu grass, with a 17.8% protein intake and T4: 70% kikuyu grass, 20% biomass and 10% cassava flour with a 15.3% protein intake. One and two-way variance analysis was made and the Pearson correlation coefficient was determined. Results. An increase was observed in the population density of viable cellulolytic bacteria (CFU/ml and B. fibrisolvens statistically significant (p<0.005 with treatment T2, in contrast to T1, T3 and T4 treatments. In addition, there was a significant increase in the concentration of vaccenic acid (mg/L in the ruminal content in Rusitec with the same treatment (T2. Conclusions. Results obtained in this ruminal simulation study are evidence to the benefits of kikuyu grass together with cassava flour and biomass diet implementation on the growth of ruminal cellulolytic and B. fibrisolvens bacteria, as well as on the production of vaccenic acid. The study also suggests the nutritional potential that such supplements could provide to grazing bovine feeding.

Thermophilic subseafloor microorganisms from the 1996 North Gorda Ridge eruption

Science.gov (United States)

Summit, Melanie; Baross, John A.

1998-12-01

High-temperature microbes were present in two hydrothermal event plumes (EP96A and B) resulting from the February-March 1996 eruptions along the North Gorda Ridge. Anaerobic thermophiles were cultured from 17 of 22 plume samples at levels exceeding 200 organisms per liter; no thermophiles were cultured from any of 12 samples of background seawater. As these microorganisms grow at temperatures of 50-90°C, they could not have grown in the event plume and instead most probably derived from a subseafloor environment tapped by the event plume source fluids. Event plumes are thought to derive from a pre-existing subseafloor fluid reservoir, which implies that these thermophiles are members of a native subseafloor community that was present before the eruptive event. Thermophiles also were cultured from continuous chronic-style hydrothermal plumes in April 1996; these plumes may have formed from cooling lava piles. To better understand the nutritional, chemical, and physical constraints of pre-eruptive crustal environments, seven coccoidal isolates from the two event plumes were partially characterized. Results from nutritional and phylogenetic studies indicate that these thermophiles are heterotrophic archaea that represent new species, and probably a new genus, within the Thermococcales.

Bioprospecting thermophiles for cellulase production: a review

Directory of Open Access Journals (Sweden)

Somen Acharya

2012-09-01

Full Text Available Most of the potential bioprospecting is currently related to the study of the extremophiles and their potential use in industrial processes. Recently microbial cellulases find applications in various industries and constitute a major group of industrial enzymes. Considerable amount of work has been done on microbial cellulases, especially with resurgence of interest in biomass ethanol production employing cellulases and use of cellulases in textile and paper industry. Most efficient method of lignocellulosic biomass hydrolysis is through enzymatic saccharification using cellulases. Significant information has also been gained about the physiology of thermophilic cellulases producers and process development for enzyme production and biomass saccharification. The review discusses the current knowledge on cellulase producing thermophilic microorganisms, their physiological adaptations and control of cellulase gene expression. It discusses the industrial applications of thermophilic cellulases, their cost of production and challenges in cellulase research especially in the area of improving process economics of enzyme production.

Comparing Residue Clusters from Thermophilic and Mesophilic Enzymes Reveals Adaptive Mechanisms.

Directory of Open Access Journals (Sweden)

Deanne W Sammond

Full Text Available Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research. Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. Thus the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions.

Relative Numbers of Certain Microbial Groups Present in Compost Used for Mushroom (Agaricus bisporus) Propagation

Science.gov (United States)

Fordyce, C.

1970-01-01

The relative numbers of microorganisms associated with compost during mushroom production were studied by the dilution plate method. Thermophilic actinomycetes and fungi were isolated with a very high frequency early in the growing season. Although numbers of thermophilic bacteria diminished slowly during the season, the thermophilic fungi and actinomycetes diminished rapidly with the latter disappearing after 6 weeks. Mesophilic fungi other than Agaricus or Trichoderma remained relatively stable throughout the growing period. Agaricus could be isolated between the first and third break. Trichoderma became dominant after the fourth break. The mesophilic bacterial counts diminished during the most productive portion of the mushroom cropping season and then increased to much higher numbers toward the end of the season. PMID:5529631

Microbiota Dynamics Associated with Environmental Conditions and Potential Roles of Cellulolytic Communities in Traditional Chinese Cereal Starter Solid-State Fermentation.

Science.gov (United States)

Li, Pan; Liang, Hebin; Lin, Wei-Tie; Feng, Feng; Luo, Lixin

2015-08-01

Traditional Chinese solid-state fermented cereal starters contain highly complex microbial communities and enzymes. Very little is known, however, about the microbial dynamics related to environmental conditions, and cellulolytic communities have never been proposed to exist during cereal starter fermentation. In this study, we performed Illumina MiSeq sequencing combined with PCR-denaturing gradient gel electrophoresis to investigate microbiota, coupled with clone library construction to trace cellulolytic communities in both fermentation stages. A succession of microbial assemblages was observed during the fermentation of starters. Lactobacillales and Saccharomycetales dominated the initial stages, with a continuous decline in relative abundance. However, thermotolerant and drought-resistant Bacillales, Eurotiales, and Mucorales were considerably accelerated during the heating stages, and these organisms dominated until the end of fermentation. Enterobacteriales were consistently ubiquitous throughout the process. For the cellulolytic communities, only the genera Sanguibacter, Beutenbergia, Agrobacterium, and Erwinia dominated the initial fermentation stages. In contrast, stages at high incubation temperature induced the appearance and dominance of Bacillus, Aspergillus, and Mucor. The enzymatic dynamics of amylase and glucoamylase also showed a similar trend, with the activities clearly increased in the first 7 days and subsequently decreased until the end of fermentation. Furthermore, β-glucosidase activity continuously and significantly increased during the fermentation process. Evidently, cellulolytic potential can adapt to environmental conditions by changes in the community structure during the fermentation of starters. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Conductive iron oxides accelerate thermophilic methanogenesis from acetate and propionate.

Science.gov (United States)

Yamada, Chihaya; Kato, Souichiro; Ueno, Yoshiyuki; Ishii, Masaharu; Igarashi, Yasuo

2015-06-01

Anaerobic digester is one of the attractive technologies for treatment of organic wastes and wastewater, while continuous development and improvements on their stable operation with efficient organic removal are required. Particles of conductive iron oxides (e.g., magnetite) are known to facilitate microbial interspecies electron transfer (termed as electric syntrophy). Electric syntrophy has been reported to enhance methanogenic degradation of organic acids by mesophilic communities in soil and anaerobic digester. Here we investigated the effects of supplementation of conductive iron oxides (magnetite) on thermophilic methanogenic microbial communities derived from a thermophilic anaerobic digester. Supplementation of magnetite accelerated methanogenesis from acetate and propionate under thermophilic conditions, while supplementation of ferrihydrite also accelerated methanogenesis from propionate. Microbial community analysis revealed that supplementation of magnetite drastically changed bacterial populations in the methanogenic acetate-degrading cultures, in which Tepidoanaerobacter sp. and Coprothermobacter sp. dominated. These results suggest that supplementation of magnetite induce electric syntrophy between organic acid-oxidizing bacteria and methanogenic archaea and accelerate methanogenesis even under thermophilic conditions. Findings from this study would provide a possibility for the achievement of stably operating thermophilic anaerobic digestion systems with high efficiency for removal of organics and generation of CH4. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass.

Science.gov (United States)

Gladden, John M; Allgaier, Martin; Miller, Christopher S; Hazen, Terry C; VanderGheynst, Jean S; Hugenholtz, Philip; Simmons, Blake A; Singer, Steven W

2011-08-15

Industrial-scale biofuel production requires robust enzymatic cocktails to produce fermentable sugars from lignocellulosic biomass. Thermophilic bacterial consortia are a potential source of cellulases and hemicellulases adapted to harsher reaction conditions than commercial fungal enzymes. Compost-derived microbial consortia were adapted to switchgrass at 60°C to develop thermophilic biomass-degrading consortia for detailed studies. Microbial community analysis using small-subunit rRNA gene amplicon pyrosequencing and short-read metagenomic sequencing demonstrated that thermophilic adaptation to switchgrass resulted in low-diversity bacterial consortia with a high abundance of bacteria related to thermophilic paenibacilli, Rhodothermus marinus, and Thermus thermophilus. At lower abundance, thermophilic Chloroflexi and an uncultivated lineage of the Gemmatimonadetes phylum were observed. Supernatants isolated from these consortia had high levels of xylanase and endoglucanase activities. Compared to commercial enzyme preparations, the endoglucanase enzymes had a higher thermotolerance and were more stable in the presence of 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), an ionic liquid used for biomass pretreatment. The supernatants were used to saccharify [C2mim][OAc]-pretreated switchgrass at elevated temperatures (up to 80°C), demonstrating that these consortia are an excellent source of enzymes for the development of enzymatic cocktails tailored to more extreme reaction conditions.

Ecology and biotechnological potential of the thermophilic fermentative Coprothermobacter spp.

Science.gov (United States)

Gagliano, M C; Braguglia, C M; Petruccioli, M; Rossetti, S

2015-05-01

Thermophilic bacteria have been isolated from several terrestrial, marine and industrial environments. Anaerobic digesters treating organic wastes are often an important source of these microorganisms, which catalyze a wide array of metabolic processes. Moreover, organic wastes are primarily composed of proteins, whose degradation is often incomplete. Coprothermobacter spp. are proteolytic anaerobic thermophilic microbes identified in several studies focused on the analysis of the microbial community structure in anaerobic thermophilic reactors. They are currently classified in the phylum Firmicutes; nevertheless, several authors showed that the Coprothermobacter group is most closely related to the phyla Dictyoglomi and Thermotoga. Since only a few proteolytic anaerobic thermophiles have been characterized so far, this microorganism has attracted the attention of researchers for its potential applications with high-temperature environments. In addition to proteolysis, Coprothermobacter spp. showed several metabolic abilities and may have a biotechnological application either as source of thermostable enzymes or as inoculum in anaerobic processes. Moreover, they can improve protein degradation by establishing a syntrophy with hydrogenotrophic archaea. To gain a better understanding of the phylogenesis, metabolic capabilities and adaptations of these microorganisms, it is of importance to better define the role in thermophilic environments and to disclose properties not yet investigated. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Browse Title Index

African Journals Online (AJOL)

Items 401 - 450 of 524 ... O.C. Ojelade, W.O. Abdul, I Abdulraheem, N.B. Ikenweiwe, A.A. Salaudeen, M.I. Akaeze. Vol 7, No 1 (2005), Review: Microbial Corrosion Of Metals, Abstract. SB Oyeleke, OA Oyewole, OP Abioye. Vol 13, No 2 (2011), Screening And Isolation Of Thermophilic Cellulolytic Bacteria From Cocoa Pod And ...

Biomass production and energy source of thermophiles in a Japanese alkaline geothermal pool.

Science.gov (United States)

Kimura, Hiroyuki; Mori, Kousuke; Nashimoto, Hiroaki; Hattori, Shohei; Yamada, Keita; Koba, Keisuke; Yoshida, Naohiro; Kato, Kenji

2010-02-01

Microbial biomass production has been measured to investigate the contribution of planktonic bacteria to fluxations in dissolved organic matter in marine and freshwater environments, but little is known about biomass production of thermophiles inhabiting geothermal and hydrothermal regions. The biomass production of thermophiles inhabiting an 85 degrees C geothermal pool was measured by in situ cultivation using diffusion chambers. The thermophiles' growth rates ranged from 0.43 to 0.82 day(-1), similar to those of planktonic bacteria in marine and freshwater habitats. Biomass production was estimated based on cellular carbon content measured directly from the thermophiles inhabiting the geothermal pool, which ranged from 5.0 to 6.1 microg C l(-1) h(-1). This production was 2-75 times higher than that of planktonic bacteria in other habitats, because the cellular carbon content of the thermophiles was much higher. Quantitative PCR and phylogenetic analysis targeting 16S rRNA genes revealed that thermophilic H2-oxidizing bacteria closely related to Calderobacterium and Geothermobacterium were dominant in the geothermal pool. Chemical analysis showed the presence of H2 in gases bubbling from the bottom of the geothermal pool. These results strongly suggested that H2 plays an important role as a primary energy source of thermophiles in the geothermal pool.

Cellulolytic enzymes, nucleic acids encoding them and methods for making and using them

Science.gov (United States)

Gray, Kevin A [San Diego, CA; Zhao, Lishan [Emeryville, CA; Cayouette, Michelle H [San Diego, CA

2012-01-24

The invention provides polypeptides having any cellulolytic activity, e.g., a cellulase activity, a endoglucanase, a cellobiohydrolase, a beta-glucosidase, a xylanase, a mannanse, a .beta.-xylosidase, an arabinofuranosidase, and/or an oligomerase activity, polynucleotides encoding these polypeptides, and methods of making and using these polynucleotides and polypeptides. In one aspect, the invention is directed to polypeptides having any cellulolytic activity, e.g., a cellulase activity, e.g., endoglucanase, cellobiohydrolase, beta-glucosidase, xylanase, mannanse, .beta.-xylosidase, arabinofuranosidase, and/or oligomerase activity, including thermostable and thermotolerant activity, and polynucleotides encoding these enzymes, and making and using these polynucleotides and polypeptides. In one aspect, the invention provides polypeptides having an oligomerase activity, e.g., enzymes that convert recalcitrant soluble oligomers to fermentable sugars in the saccharification of biomass. The polypeptides of the invention can be used in a variety of pharmaceutical, agricultural, food and feed processing and industrial contexts. The invention also provides compositions or products of manufacture comprising mixtures of enzymes comprising at least one enzyme of this invention.

Diversity of thermophilic archaeal isolates from hot springs in Japan

Science.gov (United States)

Itoh, Takashi; Yoshikawa, Naoto; Takashina, Tomonori

2005-09-01

In the light of the significance of extremophiles as model organisms to access possible extraterrestiral life, we provide a short review of the systematics of thermophilic Archaea, and introduce our exploratory research of novel thermophilic Archaea from hot springs in Japan. Up to date, we have isolated 162 strains of the thermophilic Archaea from hot springs in Japan by the enrichment method or the most probable number/PCR method, and the 16S rRNA gene sequences were determined to reveal their phylogenetic diversity. The sequence comparison illustrated that the isolates belonged to the orders Sulfolobales (117 isolates) , Thermoproteales (29 isolates), Desulfurococcales (8 isolates) and Thermoplasmatales (8 isolates), and there were six separate lineages representing new genera, and at least seven new species as predicted by the phylogenetic distance to known species. The collection of isolates not only included novel taxa but would give some implication for a necessity to reevaluate the current taxonomy of the thermophilic Archaea.

Diversity of plant oil seed-associated fungi isolated from seven oil-bearing seeds and their potential for the production of lipolytic enzymes.

Science.gov (United States)

Venkatesagowda, Balaji; Ponugupaty, Ebenezer; Barbosa, Aneli M; Dekker, Robert F H

2012-01-01

Commercial oil-yielding seeds (castor, coconut, neem, peanut, pongamia, rubber and sesame) were collected from different places in the state of Tamil Nadu (India) from which 1279 endophytic fungi were isolated. The oil-bearing seeds exhibited rich fungal diversity. High Shannon-Index H' was observed with pongamia seeds (2.847) while a low Index occurred for coconut kernel-associated mycoflora (1.018). Maximum Colonization Frequency (%) was observed for Lasiodiplodia theobromae (176). Dominance Index (expressed in terms of the Simpson's Index D) was high (0.581) for coconut kernel-associated fungi, and low for pongamia seed-borne fungi. Species Richness (Chao) of the fungal isolates was high (47.09) in the case of neem seeds, and low (16.6) for peanut seeds. All 1279 fungal isolates were screened for lipolytic activity employing a zymogram method using Tween-20 in agar. Forty isolates showed strong lipolytic activity, and were morphologically identified as belonging to 19 taxa (Alternaria, Aspergillus, Chalaropsis, Cladosporium, Colletotrichum, Curvularia, Drechslera, Fusarium, Lasiodiplodia, Mucor, Penicillium, Pestalotiopsis, Phoma, Phomopsis, Phyllosticta, Rhizopus, Sclerotinia, Stachybotrys and Trichoderma). These isolates also exhibited amylolytic, proteolytic and cellulolytic activities. Five fungal isolates (Aspergillus niger, Chalaropsis thielavioides, Colletotrichum gloeosporioides, Lasiodiplodia theobromae and Phoma glomerata) exhibited highest lipase activities, and the best producer was Lasiodiplodia theobromae (108 U/mL), which was characterized by genomic sequence analysis of the ITS region of 18S rDNA.

Production of cellulolytic enzymes by Aspergillus fumigatus

Energy Technology Data Exchange (ETDEWEB)

Trivedi, L S; Rao, K K

1979-01-01

Production of extracellular cellulase by A. fumigatus was studied in liquid shake culture. The effect of culture conditions such as C and N source and pH on cellulase was examined. Sequential appearance of cellulase components, beta-glucosidase on the 2nd day, followed by endo-beta-glucanase on the 4th day and exobeta-gluconase on the 6th day of growth was observed. Maximum production of all cellulase components was achieved on the 12th day of growth in basal medium containing cellulose as C source and a combination of (NH4)2S04 and NH4H2PO4 as N source. High cellulolytic activities were observed only in the presence of insoluble cellulose as C source, while no significant activities were observed in the presence of simple sugars.

Force-dependent melting of supercoiled DNA at thermophilic temperatures.

Science.gov (United States)

Galburt, E A; Tomko, E J; Stump, W T; Ruiz Manzano, A

2014-01-01

Local DNA opening plays an important role in DNA metabolism as the double-helix must be melted before the information contained within may be accessed. Cells finely tune the torsional state of their genomes to strike a balance between stability and accessibility. For example, while mesophilic life forms maintain negatively superhelical genomes, thermophilic life forms use unique mechanisms to maintain relaxed or even positively supercoiled genomes. Here, we use a single-molecule magnetic tweezers approach to quantify the force-dependent equilibrium between DNA melting and supercoiling at high temperatures populated by Thermophiles. We show that negatively supercoiled DNA denatures at 0.5 pN lower tension at thermophilic vs. mesophilic temperatures. This work demonstrates the ability to monitor DNA supercoiling at high temperature and opens the possibility to perform magnetic tweezers assays on thermophilic systems. The data allow for an estimation of the relative energies of base-pairing and DNA bending as a function of temperature and support speculation as to different general mechanisms of DNA opening in different environments. Lastly, our results imply that average in vivo DNA tensions range between 0.3 and 1.1 pN. Copyright © 2014 Elsevier B.V. All rights reserved.

The Cellulolytic Activity And Volatile Fatty Acid Product Of Rumen Bacteria Of Buffalo And Cattle On Rice Straw, Elephant Grass, and Sesbania Leaves Substrates

Directory of Open Access Journals (Sweden)

Caribu Hadi Prayitno

1999-01-01

Full Text Available Experiment on The Cellulolytic Activity and Volatile Fatty Acid Product of Rumen Bacteria of Buffalo and Cattle on Rice Straw, Elephant Grass, and Sesbania Leaves Substrates had been conducted at Feedstuff Laboratory of Animal Science Soedirman University. The basic design  that was used in this experiment was Completely Randomized Design (CRD with factorial pattern of 6 x 3, three replications. The bacteria isolate as the factors were cellulolytic rumen bacteria isolate of buffalo (A1, A2, and A3 and cattle (A4, A5 and A6 while the substrates (second factor  were NDF rice straw (S1, elephant grass (S2, and sesbania leaves (S3 Cell walls. The result of this experiment showed that the interaction between bacteria isolate and substrate  type were significant on pH, NDF digestibility, cellulase activity, pH was  6.28 until 6.43.  The NDF digestibility range was 12.27 until 55.61 percent. The lowers of cellulase activity was 5.11 IU/ml and the higher was 24.47 IU/ml. The range of acetic acid yield was 63.37 to 307.467 mg/100 ml. Range of  propionic production was 15.17 to 352.20 mg/ 100 ml. The production of butiric acid was 8.77 to 40.87 mg/ 100 ml. The cellulase activity  of cellulolytic rumen bacteria of buffalo was higher than cattle, and also their effect on NDF digestibility of rice straw, elephant grass, and sesbania leaves cell walls. The A3 of cellulolytic rumen bacteria isolate of  buffalo changed cell walls substrat to volatile fatty  acid was more effective than cattle, especially on cell elephant grass. Propionic and butiric  acid that was produced by cellulolytic rumen bacteria isolate of buffalo more higher than cattle (Animal Production 1 (1 : 1-9 (1999 Key Words: Cellulolytic, VFA, Rumen Bacteria, Buffalo, Cattle.

Mitigation of Membrane Biofouling in MBR Using a Cellulolytic Bacterium, Undibacterium sp. DM-1, Isolated from Activated Sludge.

Science.gov (United States)

Nahm, Chang Hyun; Lee, Seonki; Lee, Sang Hyun; Lee, Kibaek; Lee, Jaewoo; Kwon, Hyeokpil; Choo, Kwang-Ho; Lee, Jung-Kee; Jang, Jae Young; Lee, Chung-Hak; Park, Pyung-Kyu

2017-03-28

Biofilm formation on the membrane surface results in the loss of permeability in membrane bioreactors (MBRs) for wastewater treatment. Studies have revealed that cellulose is not only produced by a number of bacterial species but also plays a key role during formation of their biofilm. Hence, in this study, cellulase was introduced to a MBR as a cellulose-induced biofilm control strategy. For practical application of cellulase to MBR, a cellulolytic ( i.e ., cellulase-producing) bacterium, Undibacterium sp. DM-1, was isolated from a lab-scale MBR for wastewater treatment. Prior to its application to MBR, it was confirmed that the cell-free supernatant of DM-1 was capable of inhibiting biofilm formation and of detaching the mature biofilm of activated sludge and cellulose-producing bacteria. This suggested that cellulase could be an effective anti-biofouling agent for MBRs used in wastewater treatment. Undibacterium sp. DM-1-entrapping beads ( i.e ., cellulolytic-beads) were applied to a continuous MBR to mitigate membrane biofouling 2.2-fold, compared with an MBR with vacant-beads as a control. Subsequent analysis of the cellulose content in the biofilm formed on the membrane surface revealed that this mitigation was associated with an approximately 30% reduction in cellulose by cellulolytic-beads in MBR.

Sample handling factors affecting the enumeration of lactobacilli and cellulolytic bacteria in equine feces

Science.gov (United States)

The objectives were to compare media types and evaluate the effects of fecal storage time and temperature on the enumeration of cellulolytic bacteria and lactobacilli from horses. Fecal samples were collected from horses (n = 3) and transported to the lab (CO2, 37 ºC, 0.5 h). The samples were assign...

Cellulase and Xylanase Production from Three Isolates of Indigenous Endophytic Fungi

Science.gov (United States)

Yopi; Tasia, W.; Melliawati, R.

2017-12-01

Cellulases and hemicellulases have good potential to be used in energy production, in pulp, paper, textile industries, as well as in animal feed industries. Moreover, its utilization in food industries also cannot be ignored, among others, cellulase and xylanase roles in bakery, wine, and fruit and vegetables juice production. One of the potential enzyme source is endophytic fungi. Object of this study is to explore the potency of endophytic fungi isolated from medicinal plants as source of cellulolytic and xylanolytic enzymes. HL.47F.216 is endophytic fungi isolated from traditional medicinal plants ironwood tree was determined as xylanase producer. HL.51F.235 from pin-flower tree is cellulase producer, while CBN.6F.29 which produces both xylanase and cellulase is originated from Madagascar periwinkle. HL.47F.216 showed 2.5 cm in clear zone diameter and its xylanase activity was 0.262 U/mL with optimum condition pH 7 at 50°C. HL.51F.235 showed 2.4 cm clear zone diameter and 0.239 U/mL of cellulase activity at pH 5 and 70°C. CBN.6F.29 showed 2.8 cm and 0.394 U/mL (pH 5, 40°C) for its cellulase activity, while 2.3 cm and 0.439 U/mL (pH 8, 70°C) for its xylanase activity. Xylanase from HL.47F.216 and CBN.6F.29 showed low molecular masses of 20 kDa and 37-50 kDa, respectively. Molecular masses for cellulases from HL.51F.235 and CBN.6F.29 were 25 and 50 kDa for HL.51F.235 and 100 kDa for CBN.6F.29. Based on macroscopic and microscopic identification, fungal isolate CBN.6F.29 is a member of Class Coelomycetes, while HL.47F.216 was Acremonium sp. and HL.51F.235 was Aspergillus nigri.

Exogenous cellulases of thermophilic micromycetes. Pt. 1. Selection of producers

Energy Technology Data Exchange (ETDEWEB)

Kvesitadze, G; Kvachadze, L; Aleksidze, T; Chartishvili, D K

1986-01-01

More than 600 micromycetes - representatives of different genera have been investigated for their ability to produce exogenous cellulases. Most of the investigated cultures were found to produce these enzymes, 24 cultures being thermophilic, and 18 thermotolerant. Cellulase or its derivatives proved to be the most favourable carbon source for cellulase secretion. None of the thermophilic cultures studied manifested the ability of exogenous exoglucanase biosynthesis. Using UV-rays as mutagen, a mutant strain A. terreus T-49 has been obtained being characterized by an increased endo-glucanase and cellobiase activity, as compared to the initial strains. The cellulase preparations of thermophilic micromycetes contain one cellulasic component: endo-glucanase, or two: endo-glucanase and cellobiase.

Isolation and characterization of aerobic microorganisms with cellulolytic activity in the gut of endogeic earthworms.

Science.gov (United States)

Fujii, Katsuhiko; Ikeda, Kana; Yoshida, Seo

2012-09-01

The ability of earthworms to decompose lignocellulose involves the assistance of microorganisms in their digestive system. While many studies have revealed a diverse microbiota in the earthworm gut, including aerobic and anaerobic microorganisms, it remains unclear which of these species contribute to lignocellulose digestion. In this study, aerobic microorganisms with cellulolytic activity isolated from the gut of two endogeic earthworms, Amynthas heteropoda (Megascolecidae) and Eisenia fetida (Lumbricidae) were isolated by solid culture of gut homogenates using filter paper as a carbon source. A total of 48 strains, including four bacterial and four fungal genera, were isolated from two earthworm species. Characterization of these strains using enzyme assays showed that the most representative ones had exocellulase and xylanase activities, while some had weak laccase activity. These findings suggest that earthworms digest lignocellulose by exploiting microbial exocellulase and xylanase besides their own endocellulase. Phylogenetic analysis showed that among the cellulolytic isolates in both earthworm species Burkholderia and Chaetomium were the dominant bacterial and fungal members.

Utilization of mixed cellulolytic microbes from termite extract, elephant faecal solution and buffalo ruminal fluid to increase in vitro digestibility of King Grass

Directory of Open Access Journals (Sweden)

Agung Prabowo

2007-06-01

Full Text Available Cellulose is a compound of plant cell walls which is difficult to be degraded because it composed of glucose monomers linked by β-(1.4-bound. It will be hydrolysed by cellulase enzyme secreted by cellulolytic microbes. The effective digestion of cellulose needs high activity of cellulase enzyme. This research aims to increase in vitro king grass digestibility utilizing mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid. Twelve syringes contained gas test media were randomly divided into four treatments based on sources of microbe (SM, namely: S (SM: cattle ruminal fluid [S], RGK (SM: mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid [RGK], with composition 1 : 1 : 1, S-RGK (SM: S + RGK, with composition 1:1, and TM (without given treatment microbe. Digestibility was measured using gas test method. Average of gas production treatment of S-RGK (70.2 + 0.6 ml was higher and significantly different (P<0.01 compared to treatment of S (60.3 + 0.8 ml, RGK (40.8 + 2.3 ml, and TM (13.3 + 2.0 ml. Utilization of mixed cellulolytic microbes of termite extract, elephant faecal solution, and buffalo ruminal fluid (RGK that combined with microbes of cattle ruminal fluid (S could increase in vitro digestibility of king grass.

Experimental assessment of factors influencing dewatering properties of thermophilically digested biosolids

Energy Technology Data Exchange (ETDEWEB)

Zhou, Jianpeng; Mavinic, Donald S.; Kelly, Harlan G.; Ramey, William D.

2003-07-01

Beneficial land application of processed wastewater sludges (biosolids) is a cost-effective, and environmentally sustainable option for the final disposal of sludges, because nutrients and organic matters in the sludge are recovered and reused as a resource. Thermophilic sludge digestion produces Class A biosolids, which can be reused without restrictions. Recent experience from full-scale thermophilic sludge digestion facilities in North America revealed that, dewatering thermophilically digested biosolids required more polymers to condition than mesophilically digested biosolids. This paper reports a laboratory study that investigated factors having significant impacts on dewatering properties of digested biosolids, and assessed the relationship among digestion, dewatering properties, and characteristics of thermophilically digested biosolids. The experimental work used batch-operated, bench-scale aerobic sludge digesters. Dewaterability was measured as Capillary Suction Time (CST). The study found that feed sludge composition significantly affected dewaterability of digested sludge. Higher percentage of the secondary sludge in the feed sludge corresponded to more significant deterioration in dewaterability. The effect of thermophilic digestion temperatures on dewaterabilty was rapid, occurred within 3-hour of digestion, indicting a heat shock effect, rather than a microbiological effect. When a high shear was applied to the sludge in digesters, it resulted In a significant deterioration in dewaterability in the digested sludge. It appears there was a strong correlation between dewaterability and extracellular biopolymers. Enzymes (protease) treatment confirmed that role of extracellular proteins in affecting the dewatering properties of thermophilic biosolids, also revealed the complex nature of biopolymers' effect on dewaterability. Such effects might be due to protein-polysaccharides interactions, hydrogen bonding, or hydrophilic and hydrophobic

Diversity of Cultured Thermophilic Anaerobes in Hot Springs of Yunnan Province, China

Science.gov (United States)

Lin, L.; Lu, Y.; Dong, X.; Liu, X.; Wei, Y.; Ji, X.; Zhang, C.

2010-12-01

Thermophilic anaerobes including Archaea and Bacteria refer to those growing optimally at temperatures above 50°C and do not use oxygen as the terminal electron acceptor for growth. Study on thermophilic anaerobes will help to understand how life thrives under extreme conditions. Meanwhile thermophilic anaerobes are of importance in potential application and development of thermophilic biotechnology. We have surveyed culturable thermophilic anaerobes in hot springs (pH6.5-7.5; 70 - 94°C) in Rehai of Tengchong, Bangnazhang of Longlin, Eryuan of Dali,Yunnan, China. 50 strains in total were cultured from the hot springs water using Hungate anaerobic technique, and 30 strains were selected based on phenotypic diversity for analysis of 16S rDNA sequences. Phylogenetic analysis showed that 28 strains belonged to the members of five genera: Caldanaerobacter, Calaramator, Thermoanaerobacter, Dictyoglomus and Fervidobacterium, which formed five branches on the phylogenetic tree. Besides, 2 strains of methanogenic archaea were obtained. The majority of the isolates were the known species, however, seven strains were identified as novel species affiliated to the five genera based on the lower 16S rDNA sequence similarities (less than 93 - 97%) with the described species. This work would provide the future study on their diversity, distribution among different regions and the potential application of thermophilic enzyme. Supported by State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences(SKLMR-080605)and the Foundation of State Natural Science (30660009, 30960022, 31081220175).

Reduction of Fe(III) oxides by phylogenetically and physiologically diverse thermophilic methanogens.

Science.gov (United States)

Yamada, Chihaya; Kato, Souichiro; Kimura, Satoshi; Ishii, Masaharu; Igarashi, Yasuo

2014-09-01

Three thermophilic methanogens (Methanothermobacter thermautotrophicus, Methanosaeta thermophila, and Methanosarcina thermophila) were investigated for their ability to reduce poorly crystalline Fe(III) oxides (ferrihydrite) and the inhibitory effects of ferrihydrite on their methanogenesis. This study demonstrated that Fe(II) generation from ferrihydrite occurs in the cultures of the three thermophilic methanogens only when H2 was supplied as the source of reducing equivalents, even in the cultures of Mst. thermophila that do not grow on and produce CH4 from H2/CO2. While supplementation of ferrihydrite resulted in complete inhibition or suppression of methanogenesis by the thermophilic methanogens, ferrihydrite reduction by the methanogens at least partially alleviates the inhibitory effects. Microscopic and crystallographic analyses on the ferrihydrite-reducing Msr. thermophila cultures exhibited generation of magnetite on its cell surfaces through partial reduction of ferrihydrite. These findings suggest that at least certain thermophilic methanogens have the ability to extracellularly transfer electrons to insoluble Fe(III) compounds, affecting their methanogenic activities, which would in turn have significant impacts on materials and energy cycles in thermophilic anoxic environments. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

A family 5 β-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme features.

Science.gov (United States)

Lu, Haiqiang; Zhang, Huitu; Shi, Pengjun; Luo, Huiying; Wang, Yaru; Yang, Peilong; Yao, Bin

2013-09-01

A novel β-mannanase gene, man5XZ7, was cloned from thermophilic fungus Thielavia arenaria XZ7, and successfully expressed in Pichia pastoris. The gene (1,110 bp) encodes a 369-amino acid polypeptide with a molecular mass of approximately 40.8 kDa. The deduced sequence of Man5XZ7 consists of a putative 17-residue signal peptide and a catalytic module belonging to glycoside hydrolase (GH) family 5, and displays 76 % identity with the experimentally verified GH 5 endo-β-1,4-mannanase from Podospora anserina. Recombinant Man5XZ7 was optimally active at 75 °C and pH 5.0 and exhibited high activity at a wide temperature range (>50.0 % activity at 50-85 °C). Moreover, it had good adaptability to acidic to basic pH (>74.1 % activity at pH 4.0-7.0 and 25.6 % even at pH 9.0) and good stability from pH 3.0 to 10.0. These enzymatic properties showed that Man5XZ7 was a new thermophilic and alkali-tolerant β-mannanase. Further amino acid composition analysis indicated that Man5XZ7 has several characteristic features of thermophilic enzymes.

Cellulolytic enzyme expression and simultaneous conversion of lignocellulosic sugars into ethanol and xylitol by a new Candida tropicalis strain.

Science.gov (United States)

Mattam, Anu Jose; Kuila, Arindam; Suralikerimath, Niranjan; Choudary, Nettem; Rao, Peddy V C; Velankar, Harshad Ravindra

2016-01-01

Lignocellulosic ethanol production involves major steps such as thermochemical pretreatment of biomass, enzymatic hydrolysis of pre-treated biomass and the fermentation of released sugars into ethanol. At least two different organisms are conventionally utilized for producing cellulolytic enzymes and for ethanol production through fermentation, whereas in the present study a single yeast isolate with the capacity to simultaneously produce cellulases and xylanases and ferment the released sugars into ethanol and xylitol has been described. A yeast strain isolated from soil samples and identified as Candida tropicalis MTCC 25057 expressed cellulases and xylanases over a wide range of temperatures (32 and 42 °C) and in the presence of different cellulosic substrates [carboxymethylcellulose and wheat straw (WS)]. The studies indicated that the cultivation of yeast at 42 °C in pre-treated hydrolysate containing 0.5 % WS resulted in proportional expression of cellulases (exoglucanases and endoglucanases) at concentrations of 114.1 and 97.8 U g(-1) ds, respectively. A high xylanase activity (689.3 U g(-1) ds) was also exhibited by the yeast under similar growth conditions. Maximum expression of cellulolytic enzymes by the yeast occurred within 24 h of incubation. Of the sugars released from biomass after pretreatment, 49 g L(-1) xylose was aerobically converted into 15.8 g L(-1) of xylitol. In addition, 25.4 g L(-1) glucose released after the enzymatic hydrolysis of biomass was fermented by the same yeast to obtain an ethanol titer of 7.3 g L(-1). During the present study, a new strain of C. tropicalis was isolated and found to have potential for consolidated bioprocessing (CBP) applications. The strain could grow in a wide range of process conditions (temperature, pH) and in the presence of lignocellulosic inhibitors such as furfural, HMF and acetic acid. The new yeast produced cellulolytic enzymes over a wide temperature range and in the presence of

Community composition and cellulase activity of cellulolytic bacteria from forest soils planted with broad-leaved deciduous and evergreen trees.

Science.gov (United States)

Yang, Jiang-Ke; Zhang, Jing-Jing; Yu, Heng-Yu; Cheng, Jian-Wen; Miao, Li-Hong

2014-02-01

Cellulolytic bacteria in forest soil provide carbon sources to improve the soil fertility and sustain the nutrient balance of the forest ecological system through the decomposition of cellulosic remains. These bacteria can also be utilized for the biological conversion of biomass into renewable biofuels. In this study, the community compositions and activities of cellulolytic bacteria in the soils of forests planted with broad-leaved deciduous (Chang Qing Garden, CQG) and broad-leaved evergreen (Forest Park, FP) trees in Wuhan, China were resolved through restriction fragment length polymorphism (RFLP) and sequencing analysis of the 16S rRNA gene. All of the isolates exhibited 35 RFLP fingerprint patterns and were clustered into six groups at a similarity level of 50 %. The phylogeny analysis based on the 16S rRNA gene sequence revealed that these RFLP groups could be clustered into three phylogenetic groups and further divided into six subgroups at a higher resolution. Group I consists of isolates from Bacillus cereus, Bacillus subtilis complex (I-A) and from Paenibacillus amylolyticus-related complex (I-B) and exhibited the highest cellulase activity among all of the cellulolytic bacteria isolates. Cluster II consists of isolates belonging to Microbacterium testaceum (II-A), Chryseobacterium indoltheticum (II-B), and Flavobacterium pectinovorum and the related complex (II-C). Cluster III consists of isolates belonging to Pseudomonas putida-related species. The community shift with respect to the plant species and the soil properties was evidenced by the phylogenetic composition of the communities. Groups I-A and I-B, which account for 36.0 % of the cellulolytic communities in the CQG site, are the dominant groups (88.4 %) in the FP site. Alternatively, the ratio of the bacteria belonging to group III (P. putida-related isolates) shifted from 28.0 % in CQG to 4.0 % in FP. The soil nutrient analysis revealed that the CQG site planted with deciduous broad

Mesophilic and thermophilic activated sludge post-treatment of paper mill process water

NARCIS (Netherlands)

Vogelaar, J.C.T.; Bouwhuis, E.; Klapwijk, A.; Spanjers, H.; Lier, van J.B.

2002-01-01

Increasing system closure in paper mills and higher process water temperatures make the applicability of thermophilic treatment systems increasingly important. The use of activated sludge as a suitable thermophilic post-treatment system for anaerobically pre-treated paper process water from a paper

Thermophilic fermentative hydrogen production from starch-wastewater with bio-granules

Energy Technology Data Exchange (ETDEWEB)

Akutsu, Yohei; Harada, Hideki [Department of Civil and Environmental Engineering, Tohoku University, 6-6-06 Aoba, Sendai, Miyagi 980-8579 (Japan); Lee, Dong-Yeol [Research Center for Material Cycles and Waste Management, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506 (Japan); Chi, Yong-Zhi [Department of Environmental and Municipal Engineering, Tianjin Institute of Urban Construction, Jinjinggonglu 26, Tianjin 300384 (China); Li, Yu-You [Department of Environmental and Municipal Engineering, Tianjin Institute of Urban Construction, Jinjinggonglu 26, Tianjin 300384 (China); Department of Environmental Science, Tohoku University, 6-6-06 Aoba, Sendai, Miyagi 980-8579 (Japan); Yu, Han-Qing [School of Chemistry, University of Science and Technology of China, Hefei 230026 (China)

2009-06-15

In this study, the effects of the hydraulic retention time (HRT), pH and substrate concentration on the thermophilic hydrogen production of starch with an upflow anaerobic sludge bed (UASB) reactor were investigated. Starch was used as a sole substrate. Continuous hydrogen production was stably attained with a maximum H{sub 2} yield of 1.7 mol H{sub 2}/mol glucose. A H{sub 2}-producing thermophilic granule was successfully formed with diameter in the range of 0.5-4.0 mm with thermally pretreated methanogenic granules as the nuclei. The metabolic pathway of the granules was drastically changed at each operational parameter. The production of formic or lactic acids is an indication of the deterioration of hydrogen production for H{sub 2}-producing thermophilic granular sludge. (author)

Xylanase, CM-cellulase and avicelase production by the thermophilic fungus Sporotrichum thermophile

Energy Technology Data Exchange (ETDEWEB)

Margaritis, A; Merchant, R; Yaguchi, M

1983-01-01

When wheat straw was used as C source, S. thermophile produced large amounts of xylanase extracellularly in addition to CM-cellulase and Avicelase. These enzymes were isolated by alcohol precipitation, desalting, and column chromatography. The molecular weights were estimated to be 25,0065,000 and 84,000 for xylanase, CM-cellulase, and Avicelase, respectively. Serine and threonine were the most abundant amino acids and these enzymes are very acidic proteins.

Efficient Genome Editing of a Facultative Thermophile Using Mesophilic spCas9

NARCIS (Netherlands)

Mougiakos, Ioannis; Bosma, Elleke F.; Weenink, Koen; Vossen, Eric; Goijvaerts, Kirsten; Oost, van der John; Kranenburg, van Richard

2017-01-01

Well-developed genetic tools for thermophilic microorganisms are scarce, despite their industrial and scientific relevance. Whereas highly efficient CRISPR/Cas9-based genome editing is on the rise in prokaryotes, it has never been employed in a thermophile. Here, we apply Streptococcus pyogenes

Enrichment and identification of cellulolytic bacteria from the gastrointestinal tract of Giant African snail, Achatina fulica.

Science.gov (United States)

Pawar, Kiran D; Dar, Mudasir A; Rajput, Bharati P; Kulkarni, Girish J

2015-02-01

The cellulolytic bacterial community structure in gastrointestinal (GI) tract of Achatina fulica was studied using culture-independent and -dependent methods by enrichment in carboxymethyl cellulose (CMC). Culture-dependent method indicated that GI tract of snail was dominated by Enterobacteriaceae members. When tested for cellulase activities, all isolates obtained by culture-dependent method showed both or either of CMCase or avicelase activity. Isolate identified as Citrobacter freundii showed highest CMCase and medium avicelase activity. Sequencing of clones from the 16S rRNA gene clone library identified ten operational taxonomic units (OTUs), which were affiliated to Enterobacteriaceae of phylum Gammaproteobacteria. Of these ten OTUs, eight OTUs closely matched with Enterobacter and Klebsiella genera. The most abundant OTU allied to Klebsiella oxytoca accounted for 70 % of the total sequences. The members of Klebsiella and Enterobacter were observed by both methods indicating their dominance among the cellulolytic bacterial community in the GI tract of the snail.

Enrichment of Thermophilic Propionate-Oxidizing Bacteria in Syntrophy with Methanobacterium thermoautotrophicum or Methanobacterium thermoformicicum

OpenAIRE

Stams, Alfons J. M.; Grolle, Katja C. F.; Frijters, Carla T. M.; Van Lier, Jules B.

1992-01-01

Thermophilic propionate-oxidizing, proton-reducing bacteria were enriched from the granular methanogenic sludge of a bench-scale upflow anaerobic sludge bed reactor operated at 55°C with a mixture of volatile fatty acids as feed. Thermophilic hydrogenotrophic methanogens had a high decay rate. Therefore, stable, thermophilic propionate-oxidizing cultures could not be obtained by using the usual enrichment procedures. Stable and reproducible cultivation was possible by enrichment in hydrogen-p...

Dewaterability of thermophilically digested biosolids: effects of temperature and cellular polymeric substances

International Nuclear Information System (INIS)

Zhou, J.; Mavinic, D.S.; Kelly, H.G.; Ramey, W.D.

2002-01-01

Thermophilic processes digest sludge at high temperatures to produce Class A biosolids.Recent research work revealed that digestion temperature is the predominant factor affecting dewaterability of thermophilic biosolids. This paper presents findings of a laboratory study that investigated how various digestion temperatures affect dewaterability of digested biosolids, studied the phase partition of the substances affecting dewaterability in digested biosolids, and tested the role of cellular polymeric substances in affecting dewaterability.Secondary sludges were digested at 40-70 o C or 22 o C for up to 12 days. Centrate from thermophilically digested biosolids were treated with protease and boiling. This study found that, during the first few hours of digestion, higher temperatures resulted in more rapid and more significant deterioration in dewaterability than lower digestion temperatures. Continued digestion resulted in either improved (60 o C or 70 o C), or unchanged (40 o C or 50 o C), or gradually deteriorated dewaterability (22 o C). The substances affecting dewaterability were primarily located in the liquid phase of thermophilically digested biosolids. Boiling treatment did not result in significant changes in dewaterability. Protease treatment of the liquid phase of thermophilic biosolids improved dewaterability by 13-19%. Such an improvement confirmed the role of proteins in affecting dewaterability. (author)

Anaerobic thermophilic culture-system

Energy Technology Data Exchange (ETDEWEB)

Ljungdahl, L G; Wiegel, J K.W.

1981-04-14

A mixed culture system of Thermoanaerobacter ethanolicus and Clostridium thermocellum is employed for anaerobic, thermophilic ethanol fermentation of cellulose. By cellulase action, monosaccharides are formed which inhibit the growth of C. thermocellum, but are fermented by T. ethanolicus. Thus, at a regulated pH-value of 7.5, this mixed culture system of micro organisms results in a cellulose fermentation with a considerably higher ethanol yield.

Prospecting Agro-waste Cocktail: Supplementation for Cellulase Production by a Newly Isolated Thermophilic B. licheniformis 2D55.

Science.gov (United States)

Kazeem, Muinat Olanike; Shah, Umi Kalsom Md; Baharuddin, Azhari Samsu; AbdulRahman, Nor' Aini

2017-08-01

Bacteria isolated from thermophilic environment that can produce cellulase as well as utilise agro-waste biomass have a high potential for developing thermostable cellulase required in the biofuel industry. The cost for cellulase represents a significant challenge in converting lignocellulose to fermentable sugars for biofuel production. Among three potential bacteria examined, Bacillus licheniformis 2D55 (accession no. KT799651) was found to produce the highest cellulolytic activity (CMCase 0.33 U/mL and FPase 0.09 U/mL) at 18-24 h fermentation when grown on microcrystalline cellulose (MCC) as a carbon source in shake flask at 50 °C. Cellulase production process was further conducted on the untreated and NaOH pretreated rice straw (RS), rice husk (RH), sugarcane bagasse (BAG) and empty fruit bunch (EFB). Untreated BAG produced the highest FPase (0.160 U/mL), while the highest CMCase (0.150 U/mL) was supported on the pretreated RH. The mixture of untreated BAG and pretreated RH as agro-waste cocktail has remarkably improved CMCase (3.7- and 1.4-fold) and FPase (2.5- and 11.5-fold) compared to the untreated BAG and pretreated RH, respectively. The mechanism of cellulase production explored through SEM analysis and the location of cellulase enzymes of the isolate was also presented. Agro-waste cocktail supplementation provides an alternative method for an efficient production of cellulase.

Particle size variations of activated carbon on biofilm formation in thermophilic biohydrogen production from palm oil mill effluent

International Nuclear Information System (INIS)

Jamali, Nur Syakina; Jahim, Jamaliah Md; Isahak, Wan Nor Roslam Wan; Abdul, Peer Mohamed

2017-01-01

Highlights: • Biofilm was developed on GAC by self-attachment immobilisation. • Effect of various micropore size of GAC was evaluated in H_2 production. • Capability of attached-biofilm to degrade cellulosic fibre in POME was evaluated. • Microbial community colonized on GAC was characterised. - Abstract: In this study, we examined the formation of thermophilic microbial biofilm by self-attachment on microbial carrier of granular activated carbon (GAC) in five different micro-pore volumes 0.31, 0.41, 0.44, 0.48, and 0.50 cm"3/g. It was found that the highest hydrogen production rate of 100.8 ± 3.7 mmol H_2/l.d and yield of 1.01 ± 0.07 mol H_2/mol sugar were obtained at 0.44 cm"3/g volume size of GAC. The cellulolytic activity of attached-biofilm was further investigated using POME as a feedstock. The results showed that in all diluted POME substrate, the total sugar consumed by the microbes was found higher than that the amount of soluble monomeric sugar present in the POME medium. It is believe that the microbial biofilm was able to hydrolyse polymeric sugar of cellulosic fibre in the POME by performing enzymatic hydrolysis into simple monomeric sugar. The isolated biofilm bacteria that subjected to 16S rRNA gene analysis presented 99% high homology to the species of Thermoanaerobacterium thermosaccharolyticum which were guaranteed to perform a cellulosic degradation activity.

Thermophilic anaerobic digestion of Lurgi coal gasification wastewater in a UASB reactor

Energy Technology Data Exchange (ETDEWEB)

Wang, W.; Ma, W.C.; Han, H.J.; Li, H.Q.; Yuan, M. [Harbin Institute of Technology, Harbin (China)

2011-02-15

Lurgi coal gasification wastewater (LCGW) is a refractory wastewater, whose anaerobic treatment has been a severe problem due to its toxicity and poor biodegradability. Using a mesophilic (35 {+-} 2{sup o}C) reactor as a control, thermophilic anaerobic digestion (55 {+-} 2{sup o}C) of LCGW was investigated in a UASB reactor. After 120 days of operation, the removal of COD and total phenols by the thermophilic reactor could reach 50-55% and 50-60% respectively, at an organic loading rate of 2.5 kg COD/(m{sup 3} d) and HRT of 24h: the corresponding efficiencies were both only 20-30% in the mesophilic reactor. After thermophilic digestion, the wastewater concentrations of the aerobic effluent COD could reach below 200 mg/L compared with around 294 mg/L if mesophilic digestion was done and around 375 mg/L if sole aerobic pre-treatment was done. The results suggested that thermophilic anaerobic digestion improved significantly both anaerobic and aerobic biodegradation of LCGW.

(Hyper)thermophilic Enzymes: Production and Purification

NARCIS (Netherlands)

Falcicchio, P.; Levisson, M.; Kengen, S.W.M.; Koutsopoulos, S.

2014-01-01

The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our

Screening Cellulolytic Bacteria from the Digestive Tract Snail (Achatina fulica and Test the Ability of Cellulase Activity

Directory of Open Access Journals (Sweden)

Wijanarka Wijanarka

2016-11-01

Full Text Available On the research of enzyme production levels observed cellulase produced by bacteria in the digestive tract of the isolation of the Snail (Achatina fulica. Isolation of bacteria based on the ability of bacteria to grow on CMC media. The purpose of this study was to determine cellulase activity by cellulolytic bacteria. Some bacterial isolates were identified as cellulolytic bacteria, they were KE-B1, KE-B2, KE-B3, KE-B4, KE-B5, and KE-B6. Isolates KE-B6 was the best isolates. Furthermore KE-B6 isolates were grown on media production to determine the pattern of growth and enzyme activity. Measurement of cell growth was conducted by inoculating starter aged 22 hours at CMC production of liquid medium. Cellulase enzyme activity measurements was performed by the DNS method. The results showed that the highest activity by new isolate bacteria KE-B6 and its value of the activity of 0.4539 U/mL, growth rate (µ 0.377/hour and generation time (g 1.84 hour. This research expected cellulase of producing bacteria were easy, inexpensive and efficient. This enzyme can be used as an enzyme biolytic once expected to replace expensive commercial enzyme. The biotylic enzyme can be applied to strains improvement (protoplast fusion.How to CiteWijanarka, W., Kusdiyantini, E. & Parman, S. (2016. Screening Cellulolytic Bacteria from the Digestive Tract Snail (Achatina fulica and Test the Ability of Cellulase Activity. Biosaintifika: Journal of Biology & Biology Education, 8(3, 386-392. 

Enzyme activity screening of thermophilic bacteria isolated from Dusun Tua Hot Spring, Malaysia

Science.gov (United States)

Msarah, Marwan; Ibrahim, Izyanti; Aqma, Wan Syaidatul

2018-04-01

Thermophilic bacteria have biotechnological importance due to the availability of unique enzymes which are stable in extreme circumstances. The aim of this study includes to isolate thermophilic bacteria from hot spring and screen for important enzyme activities. Water samples from the Dusun Tua Hot Spring were collected and the physiochemical characterisation of water was measured. Eight thermophilic bacteria were isolated and determined to have at least three strong enzyme activity including protease, lipase, amylase, cellulase, pectinase and xylanase. The results showed that HuluC2 displayed all the enzyme activities and can be further studied.

Thermophilic composting of municipal solid waste

International Nuclear Information System (INIS)

Elango, D.; Thinakaran, N.; Panneerselvam, P.; Sivanesan, S.

2009-01-01

Process of composting has been developed for recycling of organic fraction of municipal solid waste (MSW). The bioreactor design was modified to reduce the composting process time. The main goal of this investigation was to find the optimal value of time period for composting of MSW in thermophilic bioreactor under aerobic condition. The temperature profiles correlated well with experimental data obtained during the maturation process. During this period biological degraders are introduced in to the reactor to accelerate the composting process. The compost materials were analyzed at various stages and the environmental parameters were considered. The final composting materials contained large organic content with in a short duration of 40 days. The quantity of volume reduction of raw MSW was 78%. The test result shows that the final compost material from the thermophilic reactor provides good humus to build up soil characteristics and some basic plant nutrients

Recent developments in the thermophilic microbiology of deep-sea hydrothermal vents.

Science.gov (United States)

Miroshnichenko, Margarita L; Bonch-Osmolovskaya, Elizaveta A

2006-04-01

The diversity of thermophilic prokaryotes inhabiting deep-sea hot vents was actively studied over the last two decades. The ever growing interest is reflected in the exponentially increasing number of novel thermophilic genera described. The goal of this paper is to survey the progress in this field made in the years 2000-2005. In this period, representatives of several new taxa of hyperthermophilic archaea were obtained from deep-sea environments. Two of these isolates had phenotypic features new for this group of organisms: the presence of an outer cell membrane (the genus Ignicoccus) and the ability to grow anaerobically with acetate and ferric iron (the genus Geoglobus). Also, our knowledge on the diversity of thermophilic bacteria from deep-sea thermal environments extended significantly. The new bacterial isolates represented diverse bacterial divisions: the phylum Aquificae, the subclass Epsilonproteobacteria, the order Thermotogales, the families Thermodesulfobacteriaceae, Deferribacteraceae, and Thermaceae, and a novel bacterial phylum represented by the genus Caldithrix. Most of these isolates are obligate or facultative lithotrophs, oxidizing molecular hydrogen in the course of different types of anaerobic respiration or microaerobic growth. The existence and significant ecological role of some of new bacterial thermophilic isolates was initially established by molecular methods.

Inactivation of Clostridium difficile in sewage sludge by anaerobic thermophilic digestion.

Science.gov (United States)

Xu, Changyun; Salsali, Hamidreza; Weese, Scott; Warriner, Keith

2016-01-01

There has been an increase in community-associated Clostridium difficile infections with biosolids derived from wastewater treatment being identified as one potential source. The current study evaluated the efficacy of thermophilic digestion in decreasing levels of C. difficile ribotype 078 associated with sewage sludge. Five isolates of C. difficile 078 were introduced (final density of 5 log CFU/g) into digested sludge and subjected to anaerobic digestion at mesophilic (36 or 42 °C) or thermophilic (55 °C) temperatures for up to 60 days. It was found that mesophilic digestion at 36 °C did not result in a significant reduction in C. difficile spore levels. In contrast, thermophilic sludge digestion reduced endospore levels at a rate of 0.19-2.68 log CFU/day, depending on the strain tested. The mechanism of lethality was indirect - by stimulating germination then inactivating the resultant vegetative cells. Acidification of sludge by adding acetic acid (6 g/L) inhibited the germination of spores regardless of the sludge digestion temperature. In conclusion, thermophilic digestion can be applied to reduce C. difficile in biosolids, thereby reducing the environmental burden of the enteric pathogen.

Characterisation of community structure of bacteria in parallel mesophilic and thermophilic pilot scale anaerobe sludge digesters.

Science.gov (United States)

Tauber, T; Berta, Brigitta; Székely, Anna J; Gyarmati, I; Kékesi, Katalin; Márialigeti, K; Tóth, Erika M

2007-03-01

The aim of the present work was to compare the microbial communities of a mesophilic and a thermophilic pilot scale anaerobe sludge digester. For studying the communities cultivation independent chemotaxonomical methods (RQ and PLFA analyses) and T-RFLP were applied. Microbial communities of the mesophilic and thermophilic pilot digesters showed considerable differences, both concerning the species present, and their abundance. A Methanosarcina sp. dominated the thermophilic, while a Methanosaeta sp. the mesophilic digester among Archaea. Species diversity of Bacteria was reduced in the thermophilic digester. Based on the quinone patterns in both digesters the dominance of sulphate reducing respiratory bacteria could be detected. The PLFA profiles of the digester communities were similar though in minor components characteristic differences were shown. Level of branched chain fatty acids is slightly lower in the thermophilic digester that reports less Gram positive bacteria. The relative ratio of fatty acids characteristic to Enterobacteriaceae, Bacteroidetes and Clostridia shows differences between the two digesters: their importance generally decreased under thermophilic conditions. The sulphate reducer marker (15:1 and 17:1) fatty acids are present in low quantity in both digesters.

Performance and microbial community analysis of two-stage process with extreme thermophilic hydrogen and thermophilic methane production from hydrolysate in UASB reactors

DEFF Research Database (Denmark)

Kongjan, Prawit; O-Thong, Sompong; Angelidaki, Irini

2011-01-01

The two-stage process for extreme thermophilic hydrogen and thermophilic methane production from wheat straw hydrolysate was investigated in up-flow anaerobic sludge bed (UASB) reactors. Specific hydrogen and methane yields of 89ml-H2/g-VS (190ml-H2/g-sugars) and 307ml-CH4/g-VS, respectively were...... energy of 13.4kJ/g-VS. Dominant hydrogen-producing bacteria in the H2-UASB reactor were Thermoanaerobacter wiegelii, Caldanaerobacter subteraneus, and Caloramator fervidus. Meanwhile, the CH4-UASB reactor was dominated with methanogens of Methanosarcina mazei and Methanothermobacter defluvii. The results...

Supplement to thermophilic hydrolysis of liquid manures. Bilag til termofil hydrolyse af gylle

Energy Technology Data Exchange (ETDEWEB)

1990-07-01

A supplement to ''Thermophilic hydrolysis of liquid manures'' which contains descriptions of testing methods and results for determining the influence of additives such as propionic acid or triolein on chemical reactions in connection with the decomposition of liquid manures under thermophilic conditions. (AB).

Extremely Thermophilic Microorganisms as Metabolic Engineering Platforms for Production of Fuels and Industrial Chemicals

Directory of Open Access Journals (Sweden)

Benjamin M Zeldes

2015-11-01

Full Text Available Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point that the use of these microorganisms as metabolic engineering platforms has become possible. While in its early days, complex metabolic pathways have been altered or engineered into recombinant extreme thermophiles, such that the production of fuels and chemicals at elevated temperatures has become possible. Not only does this expand the thermal range for industrial biotechnology, it also potentially provides biodiverse options for specific biotransformations unique to these microorganisms. The list of extreme thermophiles growing optimally between 70 and 100°C with genetic toolkits currently available includes archaea and bacteria, aerobes and anaerobes, coming from genera such as Caldicellulosiruptor, Sulfolobus, Thermotoga, Thermococcus and Pyrococcus. These organisms exhibit unusual and potentially useful native metabolic capabilities, including cellulose degradation, metal solubilization, and RuBisCO-free carbon fixation. Those looking to design a thermal bioprocess now have a host of potential candidates to choose from, each with its own advantages and challenges that will influence its appropriateness for specific applications. Here, the issues and opportunities for extremely thermophilic metabolic engineering platforms are considered with an eye towards potential technological

Extremely thermophilic microorganisms as metabolic engineering platforms for production of fuels and industrial chemicals

Science.gov (United States)

Zeldes, Benjamin M.; Keller, Matthew W.; Loder, Andrew J.; Straub, Christopher T.; Adams, Michael W. W.; Kelly, Robert M.

2015-01-01

Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point that the use of these microorganisms as metabolic engineering platforms has become possible. While in its early days, complex metabolic pathways have been altered or engineered into recombinant extreme thermophiles, such that the production of fuels and chemicals at elevated temperatures has become possible. Not only does this expand the thermal range for industrial biotechnology, it also potentially provides biodiverse options for specific biotransformations unique to these microorganisms. The list of extreme thermophiles growing optimally between 70 and 100°C with genetic toolkits currently available includes archaea and bacteria, aerobes and anaerobes, coming from genera such as Caldicellulosiruptor, Sulfolobus, Thermotoga, Thermococcus, and Pyrococcus. These organisms exhibit unusual and potentially useful native metabolic capabilities, including cellulose degradation, metal solubilization, and RuBisCO-free carbon fixation. Those looking to design a thermal bioprocess now have a host of potential candidates to choose from, each with its own advantages and challenges that will influence its appropriateness for specific applications. Here, the issues and opportunities for extremely thermophilic metabolic engineering platforms are considered with an eye toward potential technological advantages for high

Effect of storage time and temperature of equine feces on the subsequent enumeration of lactobacilli and cellulolytic bacteria

Science.gov (United States)

Cellulolytic bacteria and lactobacilli are beneficial microbes in the equine hindgut. There are several existing methodologies for the enumeration of these bacteria, which vary based on selective and differential media and sample handling procedures including storage time and temperature. The object...

Efficient utilization of xylanase and lipase producing thermophilic ...

African Journals Online (AJOL)

Efficient utilization of xylanase and lipase producing thermophilic marine actinomycetes ( Streptomyces albus and Streptomyces hygroscopicus ) in the production of ecofriendly alternative energy from waste.

Anaerobic Degradation of Marine Algae, Seagrass and Tropical Climbing Vines to Produce a Renewable Energy Source and the Analysis of Their Anaerobic Microbial Communities

Science.gov (United States)

2013-01-01

specific mechanisms and enzymes as a result of their complex structure. For these reasons there have been searches for other biomasses that may...Te’o, V., Saul, D., Morgan, H. 1999. Molecular diversity of thermophilic cellulolytic and hemicellulolytic bacteria. Microbiology Ecology, 28:99-110...lyase genes from deep-sea strains of Vibrio and Agarivorans and characterization of a new Vibrio enzyme . Marine Biotechnology, 12:526-533. Ugwu, C

Community dynamics and glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass

Energy Technology Data Exchange (ETDEWEB)

Gladden, J.M.; Allgaier, M.; Miller, C.S.; Hazen, T.C.; VanderGheynst, J.S.; Hugenholtz, P.; Simmons, B.A.; Singer, S.W.

2011-05-01

Industrial-scale biofuel production requires robust enzymatic cocktails to produce fermentable sugars from lignocellulosic biomass. Thermophilic bacterial consortia are a potential source of cellulases and hemicellulases adapted to harsher reaction conditions than commercial fungal enzymes. Compost-derived microbial consortia were adapted to switchgrass at 60 C to develop thermophilic biomass-degrading consortia for detailed studies. Microbial community analysis using small-subunit rRNA gene amplicon pyrosequencing and short-read metagenomic sequencing demonstrated that thermophilic adaptation to switchgrass resulted in low-diversity bacterial consortia with a high abundance of bacteria related to thermophilic paenibacilli, Rhodothermus marinus, and Thermus thermophilus. At lower abundance, thermophilic Chloroflexi and an uncultivated lineage of the Gemmatimonadetes phylum were observed. Supernatants isolated from these consortia had high levels of xylanase and endoglucanase activities. Compared to commercial enzyme preparations, the endoglucanase enzymes had a higher thermotolerance and were more stable in the presence of 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), an ionic liquid used for biomass pretreatment. The supernatants were used to saccharify [C2mim][OAc]-pretreated switchgrass at elevated temperatures (up to 80 C), demonstrating that these consortia are an excellent source of enzymes for the development of enzymatic cocktails tailored to more extreme reaction conditions.

Production of thermophilic and acidophilic endoglucanases by ...

African Journals Online (AJOL)

Production of thermophilic and acidophilic endoglucanases by mutant Trichoderma atroviride 102C1 using agro-industrial by-products. ... The effect of the carbon (sugarcane bagasse: SCB) and nitrogen (corn steep liquor: CSL) sources on ...

Magnesium requirement of some of the principal rumen cellulolytic bacteria.

Science.gov (United States)

Morales, M S; Dehority, B A

2014-09-01

Information available on the role of Mg for growth and cellulose degradation by rumen bacteria is both limited and inconsistent. In this study, the Mg requirements for two strains each of the cellulolytic rumen species Fibrobacter succinogenes (A3c and S85), Ruminococcus albus (7 and 8) and Ruminococcus flavefaciens (B34b and C94) were investigated. Maximum growth, rate of growth and lag time were all measured using a complete factorial design, 2(3)×6; factors were: strains (2), within species (3) and Mg concentrations (6). R. flavefaciens was the only species that did not grow when Mg was singly deleted from the media, and both strains exhibited a linear growth response to increasing Mg concentrations (PR. flavefaciens B34b was estimated as 0.54 mM; whereas the requirement for R. flavefaciens C94 was >0.82 as there was no plateau in growth. Although not an absolute requirement for growth, strains of the two other species of cellulolytic bacteria all responded to increasing Mg concentrations. For F. succinogenes S85, R. albus 7 and R. albus 8, their requirement estimated from maximum growth was 0.56, 0.52 and 0.51, respectively. A requirement for F. succinogenes A3c could not be calculated because there was no solution for contrasts. Whether R. flavefaciens had a Mg requirement for cellulose degradation was determined in NH3-free cellulose media, using a 2×4 factorial design, 2 strains and 4 treatments. Both strains of R. flavefaciens were found to have an absolute Mg requirement for cellulose degradation. Based on reported concentrations of Mg in the rumen, 1.0 to 10.1 mM, it seems unlikely that an in vivo deficiency of this element would occur.

Discrimination of thermophilic and mesophilic proteins

Directory of Open Access Journals (Sweden)

Vaisman Iosif I

2010-05-01

Full Text Available Abstract Background There is a considerable literature on the source of the thermostability of proteins from thermophilic organisms. Understanding the mechanisms for this thermostability would provide insights into proteins generally and permit the design of synthetic hyperstable biocatalysts. Results We have systematically tested a large number of sequence and structure derived quantities for their ability to discriminate thermostable proteins from their non-thermostable orthologs using sets of mesophile-thermophile ortholog pairs. Most of the quantities tested correspond to properties previously reported to be associated with thermostability. Many of the structure related properties were derived from the Delaunay tessellation of protein structures. Conclusions Carefully selected sequence based indices discriminate better than purely structure based indices. Combined sequence and structure based indices improve performance somewhat further. Based on our analysis, the strongest contributors to thermostability are an increase in ion pairs on the protein surface and a more strongly hydrophobic interior.

[Conversion of acetic acid to methane by thermophiles

Energy Technology Data Exchange (ETDEWEB)

Zinder, S.H.

1993-01-01

The primary goal of this project is to obtain a better understanding of thermophilic microorganisms which convert acetic acid to CH[sub 4]. The previous funding period represents a departure from earlier research in this laboratory, which was more physiological and ecological. The present work is centered on the biochemistry of the thermophile Methanothrix sp. strain CALS-1. this organism presents a unique opportunity, with its purity and relatively rapid growth, to do comparative biochemical studies with the other major acetotrophic genus Methanosarcina. We previously found that Methanothrix is capable of using acetate at concentrations 100 fold lower than Methanosarcina. This finding suggests that there are significant differences in the pathways of methanogenesis from acetate in the two genera.

Potential and utilization of thermophiles and thermostable enzymes in biorefining

Directory of Open Access Journals (Sweden)

Karlsson Eva

2007-03-01

Full Text Available Abstract In today's world, there is an increasing trend towards the use of renewable, cheap and readily available biomass in the production of a wide variety of fine and bulk chemicals in different biorefineries. Biorefineries utilize the activities of microbial cells and their enzymes to convert biomass into target products. Many of these processes require enzymes which are operationally stable at high temperature thus allowing e.g. easy mixing, better substrate solubility, high mass transfer rate, and lowered risk of contamination. Thermophiles have often been proposed as sources of industrially relevant thermostable enzymes. Here we discuss existing and potential applications of thermophiles and thermostable enzymes with focus on conversion of carbohydrate containing raw materials. Their importance in biorefineries is explained using examples of lignocellulose and starch conversions to desired products. Strategies that enhance thermostablity of enzymes both in vivo and in vitro are also assessed. Moreover, this review deals with efforts made on developing vectors for expressing recombinant enzymes in thermophilic hosts.

In vitro Cellulose Rich Organic Material Degradation by Cellulolytic Streptomyces albospinus (MTCC 8768

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Pinky Prasad

2012-09-01

Full Text Available Aims: Cellulosic biomass is the only foreseeable sustainable source of fuels and is also one of the dominating waste materials in nature resulting from human activities. Keeping in view the environmental problems like disposal of large volumes of cellulosic wastes and shortage of fossil fuel in the world, the main aim of the present investigation was to characterize and study the cellulolytic activity of Streptomyces albospinus (MTCC 8768, isolated from municipal wastes, on natural cellulosic substrates viz. straw powder, wood powder and finely grated vegetable peels.Methodology and Result: Stanier’s Basal broth with 100 mg of each of the substrates was inoculated separately with S. albospinus (MTCC No. 8768 and incubated at 37 °C for 8 days. The cellulosic substrates were re-weighed at an interval of 2 days and the difference between the initial weight and the final weight gave the amount of substratesdegraded by the isolate. It was observed that maximum degradation was observed in the grated vegetable peels (64 mg followed by straw powder (38 mg and wood powder (28 mg over a period of 8 days.Conclusion, significance and impact of study: By the selection of efficient cellulolytic microorganisms and cost-effective operational techniques, the production of useful end products from the biodegradation of the low cost enormous stock of cellulose in nature can be very beneficial.

NREL Explains the Higher Cellulolytic Activity of a Vital Microorganism

Energy Technology Data Exchange (ETDEWEB)

2016-06-01

The discovery of a new mode of action by C. thermocellum to convert biomass to biofuels is significant because the bacterium is already recognized as one of the most effective in the biosphere. Researchers found that, in addition to using common cellulase degradation mechanisms attached to cells, C. thermocellum also uses a new category of cell-free scaffolded enzymes. The new discovery will influence the strategies used to improve the cellulolytic activity of biomass degrading microbes going forward. Better understanding of this bacterium could lead to cheaper production of ethanol and drop-in fuels. Also, this discovery demonstrates that nature's biomass conversion behaviors are not fully understood and remain as opportunities for future microbial/enzyme engineering efforts.

Diversity and ubiquity of thermophilic methanogenic archaea in temperate anoxic soils.

Science.gov (United States)

Wu, Xiao-Lei; Friedrich, Michael W; Conrad, Ralf

2006-03-01

Temperate rice field soil from Vercelli (Italy) contains moderately thermophilic methanogens of the yet uncultivated rice cluster I (RC-I), which become prevalent upon incubation at temperatures of 45-50 degrees C. We studied whether such thermophilic methanogens were ubiquitously present in anoxic soils. Incubation of different rice field soils (from Italy, China and the Philippines) and flooded riparian soils (from the Netherlands) at 45 degrees C resulted in vigorous CH(4) production after a lag phase of about 10 days. The archaeal community structure in the soils was analysed by terminal restriction fragment length polymorphism (T-RFLP) targeting the SSU rRNA genes retrieved from the soil, and by cloning and sequencing. Clones of RC-I methanogens mostly exhibited T-RF of 393 bp, but also terminal restriction fragment (T-RF) of 158 and 258 bp length, indicating a larger diversity than previously assumed. No RC-I methanogens were initially found in flooded riparian soils. However, these archaea became abundant upon incubation of the soil at 45 degrees C. Thermophilic RC-I methanogens were also found in the rice field soils from Pavia, Pila and Gapan. However, the archaeal communities in these soils also contained other methanogenic archaea at high temperature. Rice field soil from Buggalon, on the other hand, only contained thermophilic Methanomicrobiales rather than RC-I methanogens, and rice field soil from Jurong mostly Methanomicrobiales and only a few RC-I methanogens. The archaeal community of rice field soil from Zhenjiang almost exclusively consisted of Methanosarcinaceae when incubated at high temperature. Our results show that moderately thermophilic methanogens are common in temperate soils. However, RC-I methanogens are not always dominating or ubiquitous.

Exceptional thermal stability and organic solvent tolerance of an esterase expressed from a thermophilic host

DEFF Research Database (Denmark)

Mei, Yuxia; Peng, Nan; Zhao, Shumiao

2012-01-01

, giving SisEstA. Upon Escherichia coli expression, only the thioredoxin-tagged EstA recombinant protein was soluble. The fusion protein was then purified, and removing the protein tag yielded EcSisEstA. Both forms of the thermophilic EstA enzyme were characterized. We found that SisEstA formed dimer...... that of EcSisEstA at 90°C. This indicated that thermophilic enzymes yielded from homologous expression should be better biocatalysts than those obtained from mesophilic expression.......A protein expression system recently developed for the thermophilic crenarchaeon Sulfolobus islandicus was employed to produce recombinant protein for EstA, a thermophilic esterase encoded in the same organism. Large amounts of protein were readily obtained by an affinity protein purification...

Dispersal of thermophilic Desulfotomaculum endospores into Baltic Sea sediments over thousands of years.

Science.gov (United States)

de Rezende, Júlia Rosa; Kjeldsen, Kasper Urup; Hubert, Casey R J; Finster, Kai; Loy, Alexander; Jørgensen, Bo Barker

2013-01-01

Patterns of microbial biogeography result from a combination of dispersal, speciation and extinction, yet individual contributions exerted by each of these mechanisms are difficult to isolate and distinguish. The influx of endospores of thermophilic microorganisms to cold marine sediments offers a natural model for investigating passive dispersal in the ocean. We investigated the activity, diversity and abundance of thermophilic endospore-forming sulfate-reducing bacteria (SRB) in Aarhus Bay by incubating pasteurized sediment between 28 and 85 °C, and by subsequent molecular diversity analyses of 16S rRNA and of the dissimilatory (bi)sulfite reductase (dsrAB) genes within the endospore-forming SRB genus Desulfotomaculum. The thermophilic Desulfotomaculum community in Aarhus Bay sediments consisted of at least 23 species-level 16S rRNA sequence phylotypes. In two cases, pairs of identical 16S rRNA and dsrAB sequences in Arctic surface sediment 3000 km away showed that the same phylotypes are present in both locations. Radiotracer-enhanced most probable number analysis revealed that the abundance of endospores of thermophilic SRB in Aarhus Bay sediment was ca. 10(4) per cm(3) at the surface and decreased exponentially to 10(0) per cm(3) at 6.5 m depth, corresponding to 4500 years of sediment age. Thus, a half-life of ca. 300 years was estimated for the thermophilic SRB endospores deposited in Aarhus Bay sediments. These endospores were similarly detected in the overlying water column, indicative of passive dispersal in water masses preceding sedimentation. The sources of these thermophiles remain enigmatic, but at least one source may be common to both Aarhus Bay and Arctic sediments.

γ-irradiation resistance and UV-sensitivity of extremely thermophilic archebacteria and eubacteria

International Nuclear Information System (INIS)

Kopylov, V.M.; Bonch-Osmolovskaya, E.A.; Svetlichnyi, V.A.; Miroshnichenko, M.L.; Skobkin, V.S.

1993-01-01

Cells of extremely thermophilic sulfur-dependent archebacteria Desulfurococcus amylolyticus Z533 and Thermococcus stelleri K15 are resistant to γ-irradiation. These archebacteria survive γ-irradiation at a dose of up to 5 kGy but are no longer viable after 8-9 kGy. Comparison of the survival profiles showed that archebacteria are 12 to 25 times more resistant to γ-irradiation at moderate doses (LD 50 and LD 90 ) than E. coli K12 but are 2 to 2.5 times more sensitive than D. radiodurans. γ-irradiation at a dose of 1 to 2.5 kGy killed extremely thermophilic anaerobic eubacteria Thermotoga maritima 2706 and Thermodesulfobacterium P. All extreme thermophiles studied were more sensitive to UV-irradiation than E. coli

The isolation and characterization of new C. thermocellum strains and the evaluation of multiple anaerobic digestion systems

Science.gov (United States)

Lv, Wen

The overall objective of my research was to improve the efficiencies of bioconversions that produce renewable energy from lignocellulosic biomass. To this end, my studies addressed issues important to two promising strategies: consolidated bioprocessing (CBP) and anaerobic digestion (AD). CBP achieves saccharolytic enzyme production, hydrolysis, and fermentation in a single step and is considered to be the most cost-effective model. Anaerobic bacteria that can be used in CBP are highly desirable. To that end, two thermophilic and cellulolytic bacterial strains were isolated and characterized (Chapter 3). Based on 16S rRNA gene sequence analysis, both strains CS7 and CS8 are closely related to Clostridium thermocellum ATCC 27405. However, they had significantly higher specific cellulase activities and ethanol/acetate ratios than C. thermocellum ATCC 27405. As a result, CS7 and CS8 are two new highly cellulolytic and ethanologenic C. thermocellum strains, with application potentials in research and development of CBP. As some of the most promising AD processes, two temperature-phased AD (TPAD) systems, in comparison with a thermophilic single-stage AD (TSAD) system and a mesophilic two-stage AD (MTAD) system, were studied in treating high-strength dairy cattle manure. The TPAD systems, with the thermophilic digesters acidified (AT-TPAD, Chapter 4) or operated at neutral pH (NT-TPAD, Chapter 5), were optimized at the thermophilic temperature of 50°C and a volume ratio between the thermophilic and the mesophilic digesters of 1:2. Despite similar methane productions, the NT-TPAD system achieved significantly higher volatile solid (VS) removal than the AT-TPAD system and needed no external pH adjustments (Chapter 6). At the same overall OLR, the TSAD system achieved the highest performance, followed by the NT-TPAD and the MTAD systems (Chapter 7). Each digester harbored distinct yet dynamic microbial populations, some of which were significantly correlated or associated

Microbial Consortium with High Cellulolytic Activity (MCHCA for enhanced biogas production.

Directory of Open Access Journals (Sweden)

Krzysztof ePoszytek

2016-03-01

Full Text Available The use of lignocellulosic biomass as a substrate in agricultural biogas plants is very popular and yields good results. However, the efficiency of anaerobic digestion, and thus biogas production, is not always satisfactory due to the slow or incomplete degradation (hydrolysis of plant matter. To enhance the solubilization of the lignocellulosic biomass various physical, chemical and biological pretreatment methods are used.The aim of this study was to select and characterize cellulose-degrading bacteria, and to construct a microbial consortium, dedicated for degradation of maize silage and enhancing biogas production from this substrate.Over one hundred strains of cellulose-degrading bacteria were isolated from: sewage sludge, hydrolyzer from an agricultural biogas plant, cattle slurry and manure. After physiological characterization of the isolates, sixteen strains (representatives of Bacillus, Providencia and Ochrobactrum genera were chosen for the construction of a Microbial Consortium with High Cellulolytic Activity, called MCHCA. The selected strains had a high endoglucanase activity (exceeding 0.21 IU/mL CMCase activity and a wide range of tolerance to various physical and chemical conditions. Lab-scale simulation of biogas production using the selected strains for degradation of maize silage was carried out in a two-bioreactor system, similar to those used in agricultural biogas plants.The obtained results showed that the constructed MCHCA consortium is capable of efficient hydrolysis of maize silage, and increases biogas production by even 38%, depending on the inoculum used for methane fermentation. The results in this work indicate that the mesophilic Microbial Consortium with High Cellulolytic Activity has a great potential for application on industrial scale in agricultural biogas plants.

Molecular characterization of thermophilic Campylobacter species ...

African Journals Online (AJOL)

We identified two species of thermophilic Campylobacter in companion dogs in Jos. Majority of C. jejuni were isolated from mucoid faeces while mixed infections of the two species were more common among diarrhoeic dogs. Pet owners should observe strict hand hygiene especially after handling dogs or their faeces to ...

Temporal variations in microbial biomass C and cellulolytic enzyme activity in arable soils: effects of organic matter input

DEFF Research Database (Denmark)

Debosz, K.; Rasmussen, Peter Have; Pedersen, A. R.

1999-01-01

Temporal variations in soil microbial biomass C concentration and in activity of extracellular enzymes of the cellulolytic complex were investigated in a field experiment after eight years of cultivation with either low organic matter input (low-OM) or high organic matter input (high-OM). The cul......Temporal variations in soil microbial biomass C concentration and in activity of extracellular enzymes of the cellulolytic complex were investigated in a field experiment after eight years of cultivation with either low organic matter input (low-OM) or high organic matter input (high......-OM). The cultivation systems differed in whether their source of fertiliser was mainly mineral or organic, in whether a winter cover crop was grown, and whether straw was mulched or removed. Sampling occurred at approximately monthly intervals, over a period of two years. Distinct temporal variations in microbial......) and an endocellulase activity of 44.2 +/- 1.1 nmol g(-1) h(-1). (C) 1999 Elsevier Science B.V. All rights reserved....

Biogas production and methanogenic archaeal community in mesophilic and thermophilic anaerobic co-digestion processes.

Science.gov (United States)

Yu, D; Kurola, J M; Lähde, K; Kymäläinen, M; Sinkkonen, A; Romantschuk, M

2014-10-01

Over 258 Mt of solid waste are generated annually in Europe, a large fraction of which is biowaste. Sewage sludge is another major waste fraction. In this study, biowaste and sewage sludge were co-digested in an anaerobic digestion reactor (30% and 70% of total wet weight, respectively). The purpose was to investigate the biogas production and methanogenic archaeal community composition in the anaerobic digestion reactor under meso- (35-37 °C) and thermophilic (55-57 °C) processes and an increasing organic loading rate (OLR, 1-10 kg VS m(-3) d(-1)), and also to find a feasible compromise between waste treatment capacity and biogas production without causing process instability. In summary, more biogas was produced with all OLRs by the thermophilic process. Both processes showed a limited diversity of the methanogenic archaeal community which was dominated by Methanobacteriales and Methanosarcinales (e.g. Methanosarcina) in both meso- and thermophilic processes. Methanothermobacter was detected as an additional dominant genus in the thermophilic process. In addition to operating temperatures, the OLRs, the acetate concentration, and the presence of key substrates like propionate also affected the methanogenic archaeal community composition. A bacterial cell count 6.25 times higher than archaeal cell count was observed throughout the thermophilic process, while the cell count ratio varied between 0.2 and 8.5 in the mesophilic process. This suggests that the thermophilic process is more stable, but also that the relative abundance between bacteria and archaea can vary without seriously affecting biogas production. Copyright © 2014 Elsevier Ltd. All rights reserved.

Novel thermophilic hemicellulases for the conversion of lignocellulose for second generation biorefineries.

Science.gov (United States)

Cobucci-Ponzano, Beatrice; Strazzulli, Andrea; Iacono, Roberta; Masturzo, Giuseppe; Giglio, Rosa; Rossi, Mosè; Moracci, Marco

2015-10-01

The biotransformation of lignocellulose biomasses into fermentable sugars is a very complex procedure including, as one of the most critical steps, the (hemi) cellulose hydrolysis by specific enzymatic cocktails. We explored here, the potential of stable glycoside hydrolases from thermophilic organisms, so far not used in commercial enzymatic preparations, for the conversion of glucuronoxylan, the major hemicellulose of several energy crops. Searches in the genomes of thermophilic bacteria led to the identification, efficient production, and detailed characterization of novel xylanase and α-glucuronidase from Alicyclobacillus acidocaldarius (GH10-XA and GH67-GA, respectively) and a α-glucuronidase from Caldicellulosiruptor saccharolyticus (GH67-GC). Remarkably, GH10-XA, if compared to other thermophilic xylanases from this family, coupled good specificity on beechwood xylan and the best stability at 65 °C (3.5 days). In addition, GH67-GC was the most stable α-glucuronidases from this family and the first able to hydrolyse both aldouronic acid and aryl-α-glucuronic acid substrates. These enzymes, led to the very efficient hydrolysis of beechwood xylan by using 7- to 9-fold less protein (concentrations thermophilic enzymes. In addition, remarkably, together with a thermophilic β-xylosidase, they catalyzed the production of xylose from the smart cooking pre-treated biomass of one of the most promising energy crops for second generation biorefineries. We demonstrated that search by the CAZy Data Bank of currently available genomes and detailed enzymatic characterization of recombinant enzymes allow the identification of glycoside hydrolases with novel and interesting properties and applications. Copyright © 2015 Elsevier Inc. All rights reserved.

Defluviitalea phaphyphila sp. nov., a Novel Thermophilic Bacterium That Degrades Brown Algae.

Science.gov (United States)

Ji, Shi-Qi; Wang, Bing; Lu, Ming; Li, Fu-Li

2016-02-01

Brown algae are one of the largest groups of oceanic primary producers for CO2 removal and carbon sinks for coastal regions. However, the mechanism for brown alga assimilation remains largely unknown in thermophilic microorganisms. In this work, a thermophilic alginolytic community was enriched from coastal sediment, from which an obligate anaerobic and thermophilic bacterial strain, designated Alg1, was isolated. Alg1 shared a 16S rRNA gene identity of 94.6% with Defluviitalea saccharophila LIND6LT2(T). Phenotypic, chemotaxonomic, and phylogenetic studies suggested strain Alg1 represented a novel species of the genus Defluviitalea, for which the name Defluviitalea phaphyphila sp. nov. is proposed. Alg1 exhibited an intriguing ability to convert carbohydrates of brown algae, including alginate, laminarin, and mannitol, to ethanol and acetic acid. Three gene clusters participating in this process were predicted to be in the genome, and candidate enzymes were successfully expressed, purified, and characterized. Six alginate lyases were demonstrated to synergistically deconstruct alginate into unsaturated monosaccharide, followed by one uronic acid reductase and two 2-keto-3-deoxy-d-gluconate (KDG) kinases to produce pyruvate. A nonclassical mannitol 1-phosphate dehydrogenase, catalyzing D-mannitol 1-phosphate to fructose 1-phosphate in the presence of NAD(+), and one laminarase also were disclosed. This work revealed that a thermophilic brown alga-decomposing system containing numerous novel thermophilic alginate lyases and a unique mannitol 1-phosphate dehydrogenase was adopted by the natural ethanologenic strain Alg1 during the process of evolution in hostile habitats. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Fungi and fungi-like Oomycetes isolated from affected leaves of rhododendron

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Maria Kowalik

2013-12-01

Full Text Available The aim of the work is to identify fungi and fungi-like Oomycetes occurring on affected leaves of rhododendron Rhododendron L. Mycological analyses were carried out on 200 leaves collected from green areas of Kraków from May till September 2005. Isolated fungi-like Oomycetes belonged to 67 taxa. The most frequently found fungi included: Alternaria alternata, Aspergillus niger, Botrytis cinerea, Coelophoma empetri, Nigrospora sphaerica, Pestalotia sydowiana, Phialophora cyclaminis, Phomopsis archeri, Septoria azalea and Sordaria fimicola. Among fungi-like organisms Phytophthora cinnamomi and P. citricola were isolated.

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

OpenAIRE

Aanniz,Tarik; Ouadghiri,Mouna; Melloul,Marouane; Swings,Jean; Elfahime,Elmostafa; Ibijbijen,Jamal; Ismaili,Mohamed; Amar,Mohamed

2015-01-01

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. ...

FungiDB: An Integrated Bioinformatic Resource for Fungi and Oomycetes

Directory of Open Access Journals (Sweden)

Evelina Y. Basenko

2018-03-01

Full Text Available FungiDB (fungidb.org is a free online resource for data mining and functional genomics analysis for fungal and oomycete species. FungiDB is part of the Eukaryotic Pathogen Genomics Database Resource (EuPathDB, eupathdb.org platform that integrates genomic, transcriptomic, proteomic, and phenotypic datasets, and other types of data for pathogenic and nonpathogenic, free-living and parasitic organisms. FungiDB is one of the largest EuPathDB databases containing nearly 100 genomes obtained from GenBank, Aspergillus Genome Database (AspGD, The Broad Institute, Joint Genome Institute (JGI, Ensembl, and other sources. FungiDB offers a user-friendly web interface with embedded bioinformatics tools that support custom in silico experiments that leverage FungiDB-integrated data. In addition, a Galaxy-based workspace enables users to generate custom pipelines for large-scale data analysis (e.g., RNA-Seq, variant calling, etc.. This review provides an introduction to the FungiDB resources and focuses on available features, tools, and queries and how they can be used to mine data across a diverse range of integrated FungiDB datasets and records.

Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past.

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Judit Ribera

Full Text Available A search for extremophile enzymes from ancient volcanic soils in El Hierro Island (Canary Islands, Spain allowed isolation of a microbial sporulated strain collection from which several enzymatic activities were tested. Isolates were obtained after sample cultivation under several conditions of nutrient contents and temperature. Among the bacterial isolates, supernatants from the strain designated JR3 displayed high esterase activity at temperatures ranging from 30 to 100°C, suggesting the presence of at least a hyper-thermophilic extracellular lipase. Sequence alignment of known thermophilic lipases allowed design of degenerated consensus primers for amplification and cloning of the corresponding lipase, named LipJ. However, the cloned enzyme displayed maximum activity at 30°C and pH 7, showing a different profile from that observed in supernatants of the parental strain. Sequence analysis of the cloned protein showed a pentapeptide motif -GHSMG- distinct from that of thermophilic lipases, and much closer to that of esterases. Nevertheless, the 3D structural model of LipJ displayed the same folding as that of thermophilic lipases, suggesting a common evolutionary origin. A phylogenetic study confirmed this possibility, positioning LipJ as a new member of the thermophilic family of bacterial lipases I.5. However, LipJ clusters in a clade close but separated from that of Geobacillus sp. thermophilic lipases. Comprehensive analysis of the cloned enzyme suggests a common origin of LipJ and other bacterial thermophilic lipases, and highlights the most probable divergent evolutionary pathway followed by LipJ, which during the harsh past times would have probably been a thermophilic enzyme, having lost these properties when the environment changed to more benign conditions.

Bacillus sp. JR3 esterase LipJ: A new mesophilic enzyme showing traces of a thermophilic past.

Science.gov (United States)

Ribera, Judit; Estupiñán, Mónica; Fuentes, Alba; Fillat, Amanda; Martínez, Josefina; Diaz, Pilar

2017-01-01

A search for extremophile enzymes from ancient volcanic soils in El Hierro Island (Canary Islands, Spain) allowed isolation of a microbial sporulated strain collection from which several enzymatic activities were tested. Isolates were obtained after sample cultivation under several conditions of nutrient contents and temperature. Among the bacterial isolates, supernatants from the strain designated JR3 displayed high esterase activity at temperatures ranging from 30 to 100°C, suggesting the presence of at least a hyper-thermophilic extracellular lipase. Sequence alignment of known thermophilic lipases allowed design of degenerated consensus primers for amplification and cloning of the corresponding lipase, named LipJ. However, the cloned enzyme displayed maximum activity at 30°C and pH 7, showing a different profile from that observed in supernatants of the parental strain. Sequence analysis of the cloned protein showed a pentapeptide motif -GHSMG- distinct from that of thermophilic lipases, and much closer to that of esterases. Nevertheless, the 3D structural model of LipJ displayed the same folding as that of thermophilic lipases, suggesting a common evolutionary origin. A phylogenetic study confirmed this possibility, positioning LipJ as a new member of the thermophilic family of bacterial lipases I.5. However, LipJ clusters in a clade close but separated from that of Geobacillus sp. thermophilic lipases. Comprehensive analysis of the cloned enzyme suggests a common origin of LipJ and other bacterial thermophilic lipases, and highlights the most probable divergent evolutionary pathway followed by LipJ, which during the harsh past times would have probably been a thermophilic enzyme, having lost these properties when the environment changed to more benign conditions.

Genetic Tools and Techniques for Recombinant Expression in Thermophilic Bacillaceae

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Eivind B. Drejer

2018-05-01

Full Text Available Although Escherichia coli and Bacillus subtilis are the most prominent bacterial hosts for recombinant protein production by far, additional species are being explored as alternatives for production of difficult-to-express proteins. In particular, for thermostable proteins, there is a need for hosts able to properly synthesize, fold, and excrete these in high yields, and thermophilic Bacillaceae represent one potentially interesting group of microorganisms for such purposes. A number of thermophilic Bacillaceae including B. methanolicus, B. coagulans, B. smithii, B. licheniformis, Geobacillus thermoglucosidasius, G. kaustophilus, and G. stearothermophilus are investigated concerning physiology, genomics, genetic tools, and technologies, altogether paving the way for their utilization as hosts for recombinant production of thermostable and other difficult-to-express proteins. Moreover, recent successful deployments of CRISPR/Cas9 in several of these species have accelerated the progress in their metabolic engineering, which should increase their attractiveness for future industrial-scale production of proteins. This review describes the biology of thermophilic Bacillaceae and in particular focuses on genetic tools and methods enabling use of these organisms as hosts for recombinant protein production.

Mesophilic and thermophilic anaerobic digestion of biologically pretreated abattoir wastewaters in an upflow anaerobic filter

International Nuclear Information System (INIS)

Gannoun, H.; Bouallagui, H.; Okbi, A.; Sayadi, S.; Hamdi, M.

2009-01-01

The hydrolysis pretreatment of abattoir wastewaters (AW), rich in organic suspended solids (fats and protein) was studied in static and stirred batch reactors without aeration in the presence of natural microbial population acclimated in a storage tank of AW. Microbial analysis showed that the major populations which contribute to the pretreatment of AW belong to the genera Bacillus. Contrary to the static pretreatment, the stirred conditions favoured the hydrolysis and solubilization of 80% of suspended matter into soluble pollution. The pretreated AW, in continuous stirred tank reactor (CSTR) at a hydraulic retention time (HRT) of 2 days, was fed to an upflow anaerobic filter (UAF) at an HRT of 2 days. The performance of anaerobic digestion of biologically pretreated AW was examined under mesophilic (37 deg. C) and thermophilic (55 deg. C) conditions. The shifting from a mesophilic to a thermophilic environment in the UAF was carried out with a short start-up of thermophilic condition. The UAF ran at organic loading rates (OLRs) ranging from 0.9 to 6 g COD/L d in mesophilic conditions and at OLRs from 0.9 to 9 g COD/L d in thermophilic conditions. COD removal efficiencies of 80-90% were achieved for OLRs up to 4.5 g COD/L d in mesophilic conditions, while the highest OLRs i.e. 9 g COD/L d led to efficiencies of 70-72% in thermophilic conditions. The biogas yield in thermophilic conditions was about 0.32-0.45 L biogas/g of COD removed for OLRs up to 4.5 g COD/L d. For similar OLR, the UAF in mesophilic conditions showed lower percentage of methanization. Mesophilic anaerobic digestion has been shown to destroy pathogens partially, whereas the thermophilic process was more efficient in the removal of indicator microorganisms and pathogenic bacteria at different organic loading rates.

Molecular diversity of thermophilic bacteria isolated from Pasinler hot spring (Erzurum, Turkey)

OpenAIRE

ADIGÜZEL, Ahmet; İNAN, Kadriye; ŞAHİN, Fikrettin; ARASOĞLU, Tulin; GÜLLÜCE, Medine

2011-01-01

The present study was conducted to determine the phenotypic and genotypic characterization of thermophilic bacteria isolated from Pasinler hot spring, Erzurum, Turkey. Fatty acid profiles, BOX PCR fingerprints, and 16S rDNA sequence data were used for the phenotypic and genotypic characterization of thermophilic bacteria. Totally 9 different bacterial strains were selected based on morphological, physiological, and biochemical tests. These strains were characterized by molecular tests includi...

Estimation of extracellular lipase enzyme produced by thermophilic bacillus sp. isolated from arid and semi-arid region of Rajasthan, India

OpenAIRE

Deeksha Gaur; Pankaj Kumar Jain; Yamini Singh Sisodia; Vivek Bajpai

2012-01-01

Thermophilic organisms can be defined as microorganisms which are adapted to live at high temperatures. The enzymes produce by thermophilic bacteria are capable of catalyzing biochemical reactions at high temperatures. Thermophilic bacteria are able to produce thermostable lipase enzymes capable of degradation of lipid at temperatures higher than those of mesophilic bacteria. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are quite useful in te...

Screening of complex thermophilic microbial community and ...

African Journals Online (AJOL)

Screening of complex thermophilic microbial community and application during municipal solid waste aerobic composting. ... African Journal of Biotechnology ... Complex microbial community HP83 and HC181 were applied during municipal solid waste aerobic composting that was carried out in a composting reactor under ...

Widespread Disulfide Bonding in Proteins from Thermophilic Archaea

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Julien Jorda

2011-01-01

Full Text Available Disulfide bonds are generally not used to stabilize proteins in the cytosolic compartments of bacteria or eukaryotic cells, owing to the chemically reducing nature of those environments. In contrast, certain thermophilic archaea use disulfide bonding as a major mechanism for protein stabilization. Here, we provide a current survey of completely sequenced genomes, applying computational methods to estimate the use of disulfide bonding across the Archaea. Microbes belonging to the Crenarchaeal branch, which are essentially all hyperthermophilic, are universally rich in disulfide bonding while lesser degrees of disulfide bonding are found among the thermophilic Euryarchaea, excluding those that are methanogenic. The results help clarify which parts of the archaeal lineage are likely to yield more examples and additional specific data on protein disulfide bonding, as increasing genomic sequencing efforts are brought to bear.

Widespread disulfide bonding in proteins from thermophilic archaea.

Science.gov (United States)

Jorda, Julien; Yeates, Todd O

2011-01-01

Disulfide bonds are generally not used to stabilize proteins in the cytosolic compartments of bacteria or eukaryotic cells, owing to the chemically reducing nature of those environments. In contrast, certain thermophilic archaea use disulfide bonding as a major mechanism for protein stabilization. Here, we provide a current survey of completely sequenced genomes, applying computational methods to estimate the use of disulfide bonding across the Archaea. Microbes belonging to the Crenarchaeal branch, which are essentially all hyperthermophilic, are universally rich in disulfide bonding while lesser degrees of disulfide bonding are found among the thermophilic Euryarchaea, excluding those that are methanogenic. The results help clarify which parts of the archaeal lineage are likely to yield more examples and additional specific data on protein disulfide bonding, as increasing genomic sequencing efforts are brought to bear.

Evolvability of thermophilic proteins from archaea and bacteria.

Science.gov (United States)

Takano, Kazufumi; Aoi, Atsushi; Koga, Yuichi; Kanaya, Shigenori

2013-07-16

Proteins from thermophiles possess high thermostability. The stabilization mechanisms differ between archaeal and bacterial proteins, whereby archaeal proteins are mainly stabilized via hydrophobic interactions and bacterial proteins by ion pairs. High stability is an important factor in promoting protein evolution, but the precise means by which different stabilization mechanisms affect the evolution process remain unclear. In this study, we investigated a random mutational drift of esterases from thermophilic archaea and bacteria at high temperatures. Our results indicate that mutations in archaeal proteins lead to improved function with no loss of stability, while mutant bacterial proteins are largely destabilized with decreased activity at high temperatures. On the basis of these findings, we suggest that archaeal proteins possess higher "evolvability" than bacterial proteins under temperature selection and are additionally able to evolve into eukaryotic proteins.

Fungi with multifunctional lifestyles: endophytic insect pathogenic fungi.

Science.gov (United States)

Barelli, Larissa; Moonjely, Soumya; Behie, Scott W; Bidochka, Michael J

2016-04-01

This review examines the symbiotic, evolutionary, proteomic and genetic basis for a group of fungi that occupy a specialized niche as insect pathogens as well as endophytes. We focus primarily on species in the genera Metarhizium and Beauveria, traditionally recognized as insect pathogenic fungi but are also found as plant symbionts. Phylogenetic evidence suggests that these fungi are more closely related to grass endophytes and diverged from that lineage ca. 100 MYA. We explore how the dual life cycles of these fungi as insect pathogens and endophytes are coupled. We discuss the evolution of insect pathogenesis while maintaining an endophytic lifestyle and provide examples of genes that may be involved in the transition toward insect pathogenicity. That is, some genes for insect pathogenesis may have been co-opted from genes involved in endophytic colonization. Other genes may be multifunctional and serve in both lifestyle capacities. We suggest that their evolution as insect pathogens allowed them to effectively barter a specialized nitrogen source (i.e. insects) with host plants for photosynthate. These ubiquitous fungi may play an important role as plant growth promoters and have a potential reservoir of secondary metabolites.

Chemical ecology of fungi.

Science.gov (United States)

Spiteller, Peter

2015-07-01

Fungi are widespread in nature and have conquered nearly every ecological niche. Fungi occur not only in terrestrial but also in freshwater and marine environments. Moreover, fungi are known as a rich source of secondary metabolites. Despite these facts, the ecological role of many of these metabolites is still unknown and the chemical ecology of fungi has not been investigated systematically so far. This review intends to present examples of the various chemical interactions of fungi with other fungi, plants, bacteria and animals and to give an overview of the current knowledge of fungal chemical ecology.

The genome sequences of Cellulomonas fimi and "Cellvibrio gilvus" reveal the cellulolytic strategies of two facultative anaerobes, transfer of "Cellvibrio gilvus" to the genus Cellulomonas, and proposal of Cellulomonas gilvus sp. nov.

Directory of Open Access Journals (Sweden)

Melissa R Christopherson

Full Text Available Actinobacteria in the genus Cellulomonas are the only known and reported cellulolytic facultative anaerobes. To better understand the cellulolytic strategy employed by these bacteria, we sequenced the genome of the Cellulomonas fimi ATCC 484(T. For comparative purposes, we also sequenced the genome of the aerobic cellulolytic "Cellvibrio gilvus" ATCC 13127(T. An initial analysis of these genomes using phylogenetic and whole-genome comparison revealed that "Cellvibrio gilvus" belongs to the genus Cellulomonas. We thus propose to assign "Cellvibrio gilvus" to the genus Cellulomonas. A comparative genomics analysis between these two Cellulomonas genome sequences and the recently completed genome for Cellulomonas flavigena ATCC 482(T showed that these cellulomonads do not encode cellulosomes but appear to degrade cellulose by secreting multi-domain glycoside hydrolases. Despite the minimal number of carbohydrate-active enzymes encoded by these genomes, as compared to other known cellulolytic organisms, these bacteria were found to be proficient at degrading and utilizing a diverse set of carbohydrates, including crystalline cellulose. Moreover, they also encode for proteins required for the fermentation of hexose and xylose sugars into products such as ethanol. Finally, we found relatively few significant differences between the predicted carbohydrate-active enzymes encoded by these Cellulomonas genomes, in contrast to previous studies reporting differences in physiological approaches for carbohydrate degradation. Our sequencing and analysis of these genomes sheds light onto the mechanism through which these facultative anaerobes degrade cellulose, suggesting that the sequenced cellulomonads use secreted, multidomain enzymes to degrade cellulose in a way that is distinct from known anaerobic cellulolytic strategies.

Process relevant screening of cellulolytic organisms for consolidated bioprocessing.

Science.gov (United States)

Antonov, Elena; Schlembach, Ivan; Regestein, Lars; Rosenbaum, Miriam A; Büchs, Jochen

2017-01-01

Although the biocatalytic conversion of cellulosic biomass could replace fossil oil for the production of various compounds, it is often not economically viable due to the high costs of cellulolytic enzymes. One possibility to reduce costs is consolidated bioprocessing (CBP), integrating cellulase production, hydrolysis of cellulose, and the fermentation of the released sugars to the desired product into one process step. To establish such a process, the most suitable cellulase-producing organism has to be identified. Thereby, it is crucial to evaluate the candidates under target process conditions. In this work, the chosen model process was the conversion of cellulose to the platform chemical itaconic acid by a mixed culture of a cellulolytic fungus with Aspergillus terreus as itaconic acid producer. Various cellulase producers were analyzed by the introduced freeze assay that measures the initial carbon release rate, quantifying initial cellulase activity under target process conditions. Promising candidates were then characterized online by monitoring their respiration activity metabolizing cellulose to assess the growth and enzyme production dynamics. The screening of five different cellulase producers with the freeze assay identified Trichoderma   reesei and Penicillium   verruculosum as most promising. The measurement of the respiration activity revealed a retarded induction of cellulase production for P.   verruculosum but a similar cellulase production rate afterwards, compared to T.   reesei . The freeze assay measurement depicted that P.   verruculosum reaches the highest initial carbon release rate among all investigated cellulase producers. After a modification of the cultivation procedure, these results were confirmed by the respiration activity measurement. To compare both methods, a correlation between the measured respiration activity and the initial carbon release rate of the freeze assay was introduced. The analysis revealed that the

Hydrogenomics of the extremely thermophilic bacterium Caldicellulosiruptor saccharolyticus

NARCIS (Netherlands)

Werken, van de H.J.G.; Verhaart, M.R.A.; Vanfossen, A.L.; Willquist, K.; Lewis, D.L.; Nichols, J.D.; Goorissen, H.P.; Mongodin, E.F.; Nelson, K.E.; Niel, van E.W.J.; Stams, A.J.M.; Ward, D.E.; Vos, de W.M.; Oost, van der J.; Kelly, R.M.; Kengen, S.W.M.

2008-01-01

Caldicellulosiruptor saccharolyticus is an extremely thermophilic, gram-positive anaerobe which ferments cellulose-, hemicellulose- and pectin-containing biomass to acetate, CO(2), and hydrogen. Its broad substrate range, high hydrogen-producing capacity, and ability to coutilize glucose and xylose

Cultivable microflora and endo mycorrhizas obtained in litter forest (Paramo Guerrero - Finca Puente de Tierra) Zipaquira, Colombia

International Nuclear Information System (INIS)

Bernal, Eleonora; Celis, Sebastian; Galindez, Ximena; Moratto, Claudia; Sanchez, Jimena; Garcia, Daniel

2006-01-01

The count of cultivable microorganisms (bacteria and fungi), cellulolytic microorganisms and endo mycorrhizas present in litter forest (property Puente de Tierra) in the Guerrero's moor, Colombia was made. The most frequently isolated microorganisms belonged to the staphylococcus, bacillus, pseudomonas, micrococcus, penicillium and rhodotorulagenus and cladosporium sp., which is a cellulolytic microorganism, was isolated in carboxymethyl cellulose agar. In addition eight morphotypes of endomycorrhizas were found, species of glomus and acaulospora predominated among them. This study contributes with the knowledge of the cultivable microorganisms of litter that have been little explored in moor ecosystems

Thermophilic anaerobic acetate-utilizing methanogens and their metabolism

DEFF Research Database (Denmark)

Mladenovska, Zuzana

Six strains of thermophilic anaerobic acetate-utilizing methanogens were isolated from different full-scale thermophilic biogas plants in China and Denmark. The strain isolated from the Chinese biogas plant was designated KN-6P and the isolates from the Danish full-scale biogas plants were......, utilizing the substrates acetate, methanol and methylamines but not hydrogen/carbon dioxide. Strain Methanosarcina sp. SO-2P was able to grow mixotrophically on methanol and hydrogen/carbon dioxide with methane formation from hydrogen and carbon dioxide occurring after methanol depletion. All six...... designated HG-1P, LVG-4P R1-1P, SO-2P and V-1P. The isolates were characterized morphologically and physiologically, and their immunological and phylogenetic relatedness to already known isolated strains were established. All isolated strains were identified as organisms belonging to genus Methanosarcina...

SACCHARIFICATION OF CORNCOB USING CELLULOLYTIC BACTERIA FOR BIOETHANOL PRODUCTION

Directory of Open Access Journals (Sweden)

TITI CANDRA SUNARTI

2010-08-01

Full Text Available The use of cellulose degrading enzyme (cellulases for hydrolysis of lignocellulosic material is a part of bioethanol production process. In this experiment, delignified corncob, its cellulose fraction and alpha cellulose were used as substrates to produce fermentable sugar by using three local isolates of celluloytic bacteria (C5-1, C4-4, C11-1 and Cmix ; mixed cultures of three isolates, and Saccharomyces cereviseae to produce ethanol. The results showed that all isolates of cellulolytic bacteria can grow on cellulose fraction better than on delignified corncob, and alpha cellulose. The highest hydrolytic activity produced from cellulose fraction was by isolate C4-4, which liberated 3.50 g/l of total sugar. Ethanol can be produced by mixed culture of bacteria and yeast, but because of competitive growth, the fermentation only produced 0.39-0.47 g/l of ethanol.

Structural adaptation of the subunit interface of oligomeric thermophilic and hyperthermophilic enzymes.

Science.gov (United States)

Maugini, Elisa; Tronelli, Daniele; Bossa, Francesco; Pascarella, Stefano

2009-04-01

Enzymes from thermophilic and, particularly, from hyperthermophilic organisms are surprisingly stable. Understanding of the molecular origin of protein thermostability and thermoactivity attracted the interest of many scientist both for the perspective comprehension of the principles of protein structure and for the possible biotechnological applications through application of protein engineering. Comparative studies at sequence and structure levels were aimed at detecting significant differences of structural parameters related to protein stability between thermophilic and hyperhermophilic structures and their mesophilic homologs. Comparative studies were useful in the identification of a few recurrent themes which the evolution utilized in different combinations in different protein families. These studies were mostly carried out at the monomer level. However, maintenance of a proper quaternary structure is an essential prerequisite for a functional macromolecule. At the environmental temperatures experienced typically by hyper- and thermophiles, the subunit interactions mediated by the interface must be sufficiently stable. Our analysis was therefore aimed at the identification of the molecular strategies adopted by evolution to enhance interface thermostability of oligomeric enzymes. The variation of several structural properties related to protein stability were tested at the subunit interfaces of thermophilic and hyperthermophilic oligomers. The differences of the interface structural features observed between the hyperthermophilic and thermophilic enzymes were compared with the differences of the same properties calculated from pairwise comparisons of oligomeric mesophilic proteins contained in a reference dataset. The significance of the observed differences of structural properties was measured by a t-test. Ion pairs and hydrogen bonds do not vary significantly while hydrophobic contact area increases specially in hyperthermophilic interfaces. Interface

Dispersal of thermophilic Desulfotomaculum endospores into Baltic Sea sediments over thousands of years

DEFF Research Database (Denmark)

Rezende, Julia Rosa de; Kjeldsen, Kasper Urup; Hubert, Casey RJ

2013-01-01

S rRNA and of the dissimilatory (bi)sulfite reductase (dsrAB) genes within the endospore-forming SRB genus Desulfotomaculum. The thermophilic Desulfotomaculum community in Aarhus Bay sediments consisted of at least 23 species-level 16S rRNA sequence phylotypes. In two cases, pairs of identical 16S r......RNA and dsrAB sequences in Arctic surface sediment 3000km away showed that the same phylotypes are present in both locations. Radiotracer-enhanced most probable number analysis revealed that the abundance of endospores of thermophilic SRB in Aarhus Bay sediment was ca. 104 per cm3 at the surface and decreased...... in water masses preceding sedimentation. The sources of these thermophiles remain enigmatic, but at least one source may be common to both Aarhus Bay and Arctic sediments....

High performance biological methanation in a thermophilic anaerobic trickle bed reactor.

Science.gov (United States)

Strübing, Dietmar; Huber, Bettina; Lebuhn, Michael; Drewes, Jörg E; Koch, Konrad

2017-12-01

In order to enhance energy efficiency of biological methanation of CO 2 and H 2 , this study investigated the performance of a thermophilic (55°C) anaerobic trickle bed reactor (ATBR) (58.1L) at ambient pressure. With a methane production rate of up to 15.4m 3 CH4 /(m 3 trickle bed ·d) at methane concentrations above 98%, the ATBR can easily compete with the performance of other mixed culture methanation reactors. Control of pH and nutrient supply turned out to be crucial for stable operation and was affected significantly by dilution due to metabolic water production, especially during demand-orientated operation. Considering practical applications, inoculation with digested sludge, containing a diverse biocenosis, showed high adaptive capacity due to intrinsic biological diversity. However, no macroscopic biofilm formation was observed at thermophilic conditions even after 313days of operation. The applied approach illustrates the high potential of thermophilic ATBRs as a very efficient energy conversion and storage technology. Copyright © 2017 Elsevier Ltd. All rights reserved.

Increased saccharification of kallar grass using ultrafiltrated enzyme from sporrotrichum thermophile

International Nuclear Information System (INIS)

Latif, F.; Rajoka, M.I.; Malik, K.A.

1991-01-01

The local wild type strain of sporotrichum thermophile when grown on untreated lingo cellulose was found to produce a greater level of B-glucosidase component along with other cellulase/xylanase components than most of the reported wild type potent strains. Culture filtrate obtained, when grown on 4% leptochloa fusca (kallar grass) was used as such and after concentration by ultrafiltration technique for saccharification purpose. Concentrated enzymes titre was increased to 1.2 and 4.0 U/ml for Fp-ase and B-glucosidase, respectively. There were losses in the enzyme titre obtained through ultrafiltration possibly due to adsorption on to the ultrafiltration membrane. Enzyme preparations used, saccharifide 5% kallar grass to 70, 55, 75 and 60% (theoretical basis) from cellulases of S. thermophile concentrate, dilute, T. reesei alone and in supplementation with B-glucosidase from A. niger, respectively. Analysis by HPLC revealed slightly higher glucose yield from S. thermophile enzyme preparations, whereas higher level of xylose was attained from T. reesei preparations. Rest of the sugars pooled as Oligo-sugars were found in almost similar concentrations. (author)

Extremely thermophilic microorganisms for biomass conversion: status and prospects.

Science.gov (United States)

Blumer-Schuette, Sara E; Kataeva, Irina; Westpheling, Janet; Adams, Michael Ww; Kelly, Robert M

2008-06-01

Many microorganisms that grow at elevated temperatures are able to utilize a variety of carbohydrates pertinent to the conversion of lignocellulosic biomass to bioenergy. The range of substrates utilized depends on growth temperature optimum and biotope. Hyperthermophilic marine archaea (T(opt)>or=80 degrees C) utilize alpha- and beta-linked glucans, such as starch, barley glucan, laminarin, and chitin, while hyperthermophilic marine bacteria (T(opt)>or=80 degrees C) utilize the same glucans as well as hemicellulose, such as xylans and mannans. However, none of these organisms are able to efficiently utilize crystalline cellulose. Among the thermophiles, this ability is limited to a few terrestrial bacteria with upper temperature limits for growth near 75 degrees C. Deconstruction of crystalline cellulose by these extreme thermophiles is achieved by 'free' primary cellulases, which are distinct from those typically associated with large multi-enzyme complexes known as cellulosomes. These primary cellulases also differ from the endoglucanases (referred to here as 'secondary cellulases') reported from marine hyperthermophiles that show only weak activity toward cellulose. Many extremely thermophilic enzymes implicated in the deconstruction of lignocellulose can be identified in genome sequences, and many more promising biocatalysts probably remain annotated as 'hypothetical proteins'. Characterization of these enzymes will require intensive effort but is likely to generate new opportunities for the use of renewable resources as biofuels.

Biodegradation of Palm Kernel Cake by Cellulolytic and Hemicellulolytic Bacterial Cultures through Solid State Fermentation

Directory of Open Access Journals (Sweden)

Mohamed Idris Alshelmani

2014-01-01

Full Text Available Four cellulolytic and hemicellulolytic bacterial cultures were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ and the American Type Culture Collection (ATCC. Two experiments were conducted; the objective of the first experiment was to determine the optimum time period required for solid state fermentation (SSF of palm kernel cake (PKC, whereas the objective of the second experiment was to investigate the effect of combinations of these cellulolytic and hemicellulolytic bacteria on the nutritive quality of the PKC. In the first experiment, the SSF was lasted for 12 days with inoculum size of 10% (v/w on different PKC to moisture ratios. In the second experiment, fifteen combinations were created among the four microbes with one untreated PKC as a control. The SSF lasted for 9 days, and the samples were autoclaved, dried, and analyzed for proximate analysis. Results showed that bacterial cultures produced high enzymes activities at the 4th day of SSF, whereas their abilities to produce enzymes tended to be decreased to reach zero at the 8th day of SSF. Findings in the second experiment showed that hemicellulose and cellulose was significantly P<0.05 decreased, whereas the amount of reducing sugars were significantly P<0.05 increased in the fermented PKC (FPKC compared with untreated PKC.

In vitro production of thymine dimer by ultroviolet irradiation of DNA from mesophilic and thermophilic bacteria

International Nuclear Information System (INIS)

Yein, F.S.; Stenesh, J.

1989-01-01

Thymine dimer was produced in vitro by ultraviolet irradiation of DNA, isolated from the mesophile Bacillus licheniformis and the thermophile B. stearothermophilus. Irradiation was performed at three different temperaturs (35, 45 and 55 C) and the thymine dimer was isolated and determined. An HPLC procedure was developed that permitted temperature was greater for the thermophile than for the mesophile. Formation of thymine dimer increased with temperature for both organisms but more so for the thermophile; over the temperature range of 35-55 C, the average increase in thymine dimer production for the themrophile was about 4-times that for the mesophile. The melting out temperature, as a function of increasing irradiation temperature, was essentially unchanged for the mesophilic DNA, but decreased progressively for the thermophilic DNA. These results are discussed in terms of the macromolecular theory of to the macromolecular theory of the thermophily. (author). 31 refs.; 4 figs.; 3 tabs

State of the art review of biofuels production from lignocellulose by thermophilic bacteria.

Science.gov (United States)

Jiang, Yujia; Xin, Fengxue; Lu, Jiasheng; Dong, Weiliang; Zhang, Wenming; Zhang, Min; Wu, Hao; Ma, Jiangfeng; Jiang, Min

2017-12-01

Biofuels, including ethanol and butanol, are mainly produced by mesophilic solventogenic yeasts and Clostridium species. However, these microorganisms cannot directly utilize lignocellulosic materials, which are abundant, renewable and non-compete with human demand. More recently, thermophilic bacteria show great potential for biofuels production, which could efficiently degrade lignocellulose through the cost effective consolidated bioprocessing. Especially, it could avoid contamination in the whole process owing to its relatively high fermentation temperature. However, wild types thermophiles generally produce low levels of biofuels, hindering their large scale production. This review comprehensively summarizes the state of the art development of biofuels production by reported thermophilic microorganisms, and also concludes strategies to improve biofuels production including the metabolic pathways construction, co-culturing systems and biofuels tolerance. In addition, strategies to further improve butanol production are proposed. Copyright © 2017 Elsevier Ltd. All rights reserved.

Methanomethylovorans thermophila sp. nov., a thermophilic, methylotrophic methanogen form an anaerobic reactor fed with methanol

NARCIS (Netherlands)

Jiang, B.; Parshina, S.N.; Doesburg, van W.C.J.; Lomans, B.P.; Stams, A.J.M.

2005-01-01

A novel thermophilic, obligately methylotrophic, methanogenic archaeon, strain L2FAWT, was isolated from a thermophilic laboratory-scale upflow anaerobic sludge blanket reactor fed with methanol as the carbon and energy source. Cells of strain L2FAWT were non-motile, irregular cocci, 0·7¿1·5 µm in

Up-Streaming Process for Glucose Oxidase by Thermophilic Penicillium sp. in Shake Flask

OpenAIRE

Muhammad Mohsin JAVED; Aroosh SHABIR; Sana ZAHOOR; Ikram UL-HAQ

2012-01-01

The present study is concerned with the production of glucose oxidase (GOD) from thermophilic Penicillium sp. in 250 mL shake flask. Fourteen different strains of thermophilic Penicillium sp. were isolated from the soil and were screened for glucose oxidase production. IIBP-13 strain gave maximum extra-cellular glucose oxidase production as compared to other isolates. Effect of submerged fermentation in shaking and static conditions, different carbon sources and incubation period on the produ...

Rapid startup of thermophilic anaerobic digester to remove tetracycline and sulfonamides resistance genes from sewage sludge.

Science.gov (United States)

Xu, Rui; Yang, Zhao-Hui; Wang, Qing-Peng; Bai, Yang; Liu, Jian-Bo; Zheng, Yue; Zhang, Yan-Ru; Xiong, Wei-Ping; Ahmad, Kito; Fan, Chang-Zheng

2018-01-15

Spread of antibiotic resistance genes (ARGs) originating from sewage sludge is highlighted as an eminent health threat. This study established a thermophilic anaerobic digester using one-step startup strategy to quickly remove tetracycline and sulfonamides resistance genes from sewage sludge. At least 20days were saved in the startup period from mesophilic to thermophilic condition. Based on the results of 16S rDNA amplicons sequencing and predicted metagenomic method, the successful startup largely relied on the fast colonization of core thermophilic microbial population (e.g. Firmicutes, Proteobacteria, Actinobacteria). Microbial metabolic gene pathways for substrate degradation and methane production was also increased by one-step mode. In addition, real-time quantitative PCR approach revealed that most targeted tetracycline and sulfonamides resistance genes ARGs (sulI, tetA, tetO, tetX) were substantially removed during thermophilic digestion (removal efficiency>80%). Network analysis showed that the elimination of ARGs was attributed to the decline of their horizontal (intI1 item) and vertical (potential hosts) transfer-related elements under high-temperature. This research demonstrated that rapid startup thermophilic anaerobic digestion of wastewater solids would be a suitable technology for reducing quantities of various ARGs. Copyright © 2017 Elsevier B.V. All rights reserved.

Startup and stability of thermophilic anaerobic digestion of OFMSW

KAUST Repository

El-Fadel, Mutasem E.; Saikaly, Pascal; Ghanimeh, Sophia A.

2013-01-01

Anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW) is promoted as an energy source and more recently as a greenhouse gas (GHG) mitigation measure. In this context, AD systems operating at thermophilic temperatures (55-60°C)-compared to mesophilic temperatures (35-40°C)-have the unique feature of producing hygienic soil conditioners with greater process efficiency, higher energy yield, and more GHG savings. Startup of AD systems is often constrained by the lack of acclimated seeds, leading to process instability and failure. The authors focus on strategies to startup thermophilic digesters treating OFMSW in the absence of acclimated seeds and examines constraints associated with process stability and ways to overcome them. Relevant gaps in the literature and future research needs are delineated. © 2013 Taylor & Francis Group, LLC.

Kinetics of Butyrate, Acetate, and Hydrogen Metabolism in a Thermophilic, Anaerobic, Butyrate-Degrading Triculture

OpenAIRE

Ahring, Birgitte K.; Westermann, Peter

1987-01-01

Kinetics of butyrate, acetate, and hydrogen metabolism were determined with butyrate-limited, chemostat-grown tricultures of a thermophilic butyrate-utilizing bacterium together with Methanobacterium thermoautotrophicum and the TAM organism, a thermophilic acetate-utilizing methanogenic rod. Kinetic parameters were determined from progress curves fitted to the integrated form of the Michaelis-Menten equation. The apparent half-saturation constants, Km, for butyrate, acetate, and dissolved hyd...

ISOLASI, SKRINING DAN IDENTIFIKASI JAMUR XILANOLITIK LOKAL YANG BERPOTENSI SEBAGAI AGENSIA PEMUTIH PULP YANG RAMAH LINGKUNGAN (Isolation, Screening and Identification Xylanolytic Local Fungi that Potentially as Pulp Bleaching Agents

Directory of Open Access Journals (Sweden)

Elisa Nurnawati

2015-01-01

Xylanase has great potential for industry application. Application of xylanase can be done in pretreatment of pulp bleaching in the pulp and paper industry. Enzyme application can reduce the use of chlorine compounds that are harmful to the environment. Therefore, xylanase that used in pulp bleaching should be free of cellulase activity. Fungi are one of the groups of microbes that are able to produce xylanase. The aims of this study was to obtain local xylanase-producing fungal isolates from soil that assumed contain of xylan. The source of fungal isolates were the soil around the pulp and paper industry; Acacia forests in the district Ogan Ilir and Muara Enim, South Sumatra; Wanagama, Yogyakarta; sawmills in Palembang and Yogyakarta; and Palembang landfill. Based on the initial screening in the agar basal medium, 111 fungal isolates were obtained. Most of them were the xylanase-producing fungi, but only 12 fungal isolates that have high xylanolytic capabilities. Further screening was performed on xylan liquid basal medium. The results showed that the fungus identified as Chaetomium globosum, Penicillium simplicissimum, Aspergillus tamarii and Monocillium have higher xylanase specific activity than the other isolates. They were also have lignolytic and cellulolytic activities. Therefore, fungal xylanase potentially developed as a pulp bleaching agent.

Isolation, screening, and identification of potential cellulolytic and xylanolytic producers for biodegradation of untreated oil palm trunk and its application in saccharification of lemongrass leaves.

Science.gov (United States)

Ang, S K; Yahya, Adibah; Abd Aziz, Suraini; Md Salleh, Madihah

2015-01-01

This study presents the isolation and screening of fungi with excellent ability to degrade untreated oil palm trunk (OPT) in a solid-state fermentation system (SSF). Qualitative assay of cellulases and xylanase indicates notable secretion of both enzymes by 12 fungal strains from a laboratory collection and 5 strains isolated from a contaminated wooden board. High production of these enzymes was subsequently quantified in OPT in SSF. Aspergillus fumigates SK1 isolated from cow dung gives the highest xylanolytic activity (648.448 U g(-1)), generally high cellulolytic activities (CMCase: 48.006, FPase: 6.860, beta-glucosidase: 16.328 U g(-1)) and moderate lignin peroxidase activity (4.820 U/g), and highest xylanolytic activity. The xylanase encoding gene of Aspergillus fumigates SK1 was screened using polymerase chain reaction by a pair of degenerate primers. Through multiple alignment of the SK1 strain's xylanase nucleotide sequences with other published xylanases, it was confirmed that the gene belonged to the xylanase glycoside hydrolase family 11 (GH11) with a protein size of 24.49 kD. Saccharification of lemongrass leaves using crude cellulases and xylanase gives the maximum reducing sugars production of 6.84 g/L with glucose as the major end product and traces of phenylpropanic compounds (vanillic acid, p-coumaric acid, and ferulic acid).

Differences in the catalytic mechanisms of mesophilic and thermophilic indole-3-glycerol phosphate synthase enzymes at their adaptive temperatures.

Science.gov (United States)

Zaccardi, Margot J; Mannweiler, Olga; Boehr, David D

2012-02-10

Thermophilic enzymes tend to be less catalytically-active at lower temperatures relative to their mesophilic counterparts, despite having very similar crystal structures. An often cited hypothesis for this general observation is that thermostable enzymes have evolved a more rigid tertiary structure in order to cope with their more extreme, natural environment, but they are also less flexible at lower temperatures, leading to their lower catalytic activity under mesophilic conditions. An alternative hypothesis, however, is that complementary thermophilic-mesophilic enzyme pairs simply operate through different evolutionary-optimized catalytic mechanisms. In this communication, we present evidence that while the steps of the catalytic mechanisms for mesophilic and thermophilic indole-3-glycerol phosphate synthase (IGPS) enzymes are fundamentally similar, the identity of the rate-determining step changes as a function of temperature. Our findings indicate that while product release is rate-determining at 25°C for thermophilic IGPS, near its adaptive temperature (75°C), a proton transfer event, involving a general acid, becomes rate-determining. The rate-determining steps for thermophilic and mesophilic IGPS enzymes are also different at their respective, adaptive temperatures with the mesophilic IGPS-catalyzed reaction being rate-limited before irreversible CO2 release, and the thermophilic IGPS-catalyzed reaction being rate limited afterwards. Copyright © 2012 Elsevier Inc. All rights reserved.

Isolation and characterization of efficient cellulolytic fungi from ...

African Journals Online (AJOL)

user

potentially sustainable approach to develop novel bioprocesses and products. Microbial cellulases have become the focal biocatalysts due to their complex nature and wide spread industrial applications (Kuhad et al.,. 2011). The search for efficient microorganisms which can produce all the three types of cellulases that can ...

PCR detection of thermophilic spore-forming bacteria involved in canned food spoilage.

Science.gov (United States)

Prevost, S; Andre, S; Remize, F

2010-12-01

Thermophilic bacteria that form highly heat-resistant spores constitute an important group of spoilage bacteria of low-acid canned food. A PCR assay was developed in order to rapidly trace these bacteria. Three PCR primer pairs were designed from rRNA gene sequences. These primers were evaluated for the specificity and the sensitivity of detection. Two primer pairs allowed detection at the species level of Geobacillus stearothermophilus and Moorella thermoacetica/thermoautrophica. The other pair allowed group-specific detection of anaerobic thermophilic bacteria of the genera Thermoanaerobacterium, Thermoanaerobacter, Caldanerobium and Caldanaerobacter. After a single enrichment step, these PCR assays allowed the detection of 28 thermophiles from 34 cans of spoiled low-acid food. In addition, 13 ingredients were screened for the presence of these bacteria. This PCR assay serves as a detection method for strains able to spoil low-acid canned food treated at 55°C. It will lead to better reactivity in the canning industry. Raw materials and ingredients might be qualified not only for quantitative spore contamination, but also for qualitative contamination by highly heat-resistant spores.

Microbial conversion of food wastes for biofertilizer production with thermophilic lipolytic microbes

Energy Technology Data Exchange (ETDEWEB)

Tsai, Shu-Hsien; Yang, Shang-Shyng [Institute of Microbiology and Biochemistry, National Taiwan University, Taipei 10617, (Taiwan); Liu, Ching-Piao [Department of Biological Science and Technology, Meiho Institute of Technology, Pingtung 91201, (Taiwan)

2007-05-15

Food waste is approximately one quarter of the total garbage in Taiwan. To investigate the feasibility of microbial conversion of food waste to multiple functional biofertilizer, food waste was mixed with bulking materials, inoculated with thermophilic and lipolytic microbes and incubated at 50{sup o}C in a mechanical composter. Microbial inoculation enhanced the degradation of food wastes, increased the total nitrogen and the germination rate of alfalfa seed, shortened the maturity period and improved the quality of biofertilizer. In food waste inoculated with thermophilic and lipolytic Brevibacillus borstelensis SH168 for 28 days, total nitrogen increased from 2.01% to 2.10%, ash increased from 24.94% to 29.21%, crude fat decreased from 4.88% to 1.34% and the C/N ratio decreased from 18.02 to 17.65. Each gram of final product had a higher population of thermophilic microbes than mesophilic microbes. Microbial conversion of food waste to biofertilizer is a feasible and potential technology in the future to maintain the natural resources and to reduce the impact on environmental quality. (author)

Consistent mutational paths predict eukaryotic thermostability

Directory of Open Access Journals (Sweden)

van Noort Vera

2013-01-01

Full Text Available Abstract Background Proteomes of thermophilic prokaryotes have been instrumental in structural biology and successfully exploited in biotechnology, however many proteins required for eukaryotic cell function are absent from bacteria or archaea. With Chaetomium thermophilum, Thielavia terrestris and Thielavia heterothallica three genome sequences of thermophilic eukaryotes have been published. Results Studying the genomes and proteomes of these thermophilic fungi, we found common strategies of thermal adaptation across the different kingdoms of Life, including amino acid biases and a reduced genome size. A phylogenetics-guided comparison of thermophilic proteomes with those of other, mesophilic Sordariomycetes revealed consistent amino acid substitutions associated to thermophily that were also present in an independent lineage of thermophilic fungi. The most consistent pattern is the substitution of lysine by arginine, which we could find in almost all lineages but has not been extensively used in protein stability engineering. By exploiting mutational paths towards the thermophiles, we could predict particular amino acid residues in individual proteins that contribute to thermostability and validated some of them experimentally. By determining the three-dimensional structure of an exemplar protein from C. thermophilum (Arx1, we could also characterise the molecular consequences of some of these mutations. Conclusions The comparative analysis of these three genomes not only enhances our understanding of the evolution of thermophily, but also provides new ways to engineer protein stability.

Previously unclassified bacteria dominate during thermophilic and mesophilic anaerobic pre-treatment of primary sludge.

Science.gov (United States)

Pervin, Hasina M; Batstone, Damien J; Bond, Philip L

2013-06-01

Thermophilic biological pre-treatment enables enhanced anaerobic digestion for treatment of wastewater sludges but, at present, there is limited understanding of the hydrolytic-acidogenic microbial composition and its contribution to this process. In this study, the process was assessed by comparing the microbiology of thermophilic (50-65 °C) and mesophilic (35 °C) pre-treatment reactors treating primary sludge. A full-cycle approach for the 16S rRNA genes was applied in order to monitor the diversity of bacteria and their abundance in a thermophilic pre-treatment reactor treating primary sludge. For the thermophilic pre-treatment (TP), over 90% of the sequences were previously undetected and these had less than 97% sequence similarity to cultured organisms. During the first 83 days, members of the Betaproteobacteria dominated the community sequences and a newly designed probe was used to monitor a previously unknown bacterium affiliated with the genus Brachymonas. Between days 85 and 183, three phylotypes that affiliated with the genera Comamonas, Clostridium and Lysobacter were persistently dominant in the TP community, as revealed by terminal-restriction fragment length polymorphism (T-RFLP). Hydrolytic and fermentative functions have been speculated for these bacteria. Mesophilic pre-treatment (MP) and TP communities were different but they were both relatively dynamic. Statistical correlation analysis and the function of closely allied reference organisms indicated that previously unclassified bacteria dominated the TP community and may have been functionally involved in the enhanced hydrolytic performance of thermophilic anaerobic pre-treatment. This study is the first to reveal the diversity and dynamics of bacteria during anaerobic digestion of primary sludge. Copyright © 2013 Elsevier GmbH. All rights reserved.

Nematode-Trapping Fungi.

Science.gov (United States)

Jiang, Xiangzhi; Xiang, Meichun; Liu, Xingzhong

2017-01-01

Nematode-trapping fungi are a unique and intriguing group of carnivorous microorganisms that can trap and digest nematodes by means of specialized trapping structures. They can develop diverse trapping devices, such as adhesive hyphae, adhesive knobs, adhesive networks, constricting rings, and nonconstricting rings. Nematode-trapping fungi have been found in all regions of the world, from the tropics to Antarctica, from terrestrial to aquatic ecosystems. They play an important ecological role in regulating nematode dynamics in soil. Molecular phylogenetic studies have shown that the majority of nematode-trapping fungi belong to a monophyletic group in the order Orbiliales (Ascomycota). Nematode-trapping fungi serve as an excellent model system for understanding fungal evolution and interaction between fungi and nematodes. With the development of molecular techniques and genome sequencing, their evolutionary origins and divergence, and the mechanisms underlying fungus-nematode interactions have been well studied. In recent decades, an increasing concern about the environmental hazards of using chemical nematicides has led to the application of these biological control agents as a rapidly developing component of crop protection.

Survival of weed seeds and animal parasites as affected by anaerobic digestion at meso- and thermophilic conditions

DEFF Research Database (Denmark)

Johansen, Anders; Bangsø Nielsen, Henrik; Hansen, Christian M.

2013-01-01

did not affect egg survival during the first 48h and it took up to 10days before total elimination was reached. In general, anaerobic digestion in biogas plants seems an efficient way (thermophilic more efficient than mesophilic) to treat organic farm wastes in a way that suppresses animal parasites......, Ascaris suum, was assessed under conditions similar to biogas plants managed at meso- (37°C) and thermophilic (55°C) conditions. Cattle manure was used as digestion substrate and experimental units were sampled destructively over time. Regarding weed seeds, the effect of thermophilic conditions (55°C...

Optimization of micronutrient supplement for enhancing biogas production from food waste in two-phase thermophilic anaerobic digestion.

Science.gov (United States)

Menon, Ajay; Wang, Jing-Yuan; Giannis, Apostolos

2017-01-01

The aim of this study was to enhance the biogas productivity of two-phase thermophilic anaerobic digestion (AD) using food waste (FW) as the primary substrate. The influence of adding four trace metals (Ca, Mg, Co, and Ni) as micronutrient supplement in the methanogenic phase of the thermophilic system was investigated. Initially, Response Surface Methodology (RSM) was applied to determine the optimal concentration of micronutrients in batch experiments. The results showed that optimal concentrations of 303, 777, 7 and 3mg/L of Ca, Mg, Co and Ni, respectively, increased the biogas productivity as much as 50% and significantly reduced the processing time. The formulated supplement was tested in continuous two-phase thermophilic AD system with regard to process stability and productivity. It was found that a destabilized thermophilic AD process encountering high VFA accumulation recovered in less than two weeks, while the biogas production was improved by 40% yielding 0.46L CH 4 /gVS added /day. There was also a major increase in soluble COD utilization upon the addition of micronutrient supplement. The results of this study indicate that a micronutrient supplement containing Ca, Mg, Co and Ni could probably remedy any type of thermophilic AD process. Copyright © 2016 Elsevier Ltd. All rights reserved.

Enzymatic activity of the cellulolytic complex produced by trichoderma reesei. Enzymatic hydrolysis of cellulose

International Nuclear Information System (INIS)

Alfonsel Jaen, M.; Negro, M.J.; Saez, R.; Martin Moreno, C.

1986-01-01

The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reese QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass from Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars productions, have been selected. Previous studies on enzymatic hydrolysis of O. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (author). 10 figs.; 10 refs

Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose

International Nuclear Information System (INIS)

Alfonsel J, M.; Negro A, M. J.; Saez A, R.; Martin M, C.

1986-01-01

The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs

Fusarium and other opportunistic hyaline fungi

Science.gov (United States)

This chapter focuses on those fungi that grow in tissue in the form of hyaline or lightly colored septate hyphae. These fungi include Fusarium and other hyaline fungi. Disease caused by hyaline fungi is referred to as hyalohyphomycosis. Hyaline fungi described in this chapter include the anamorphic,...

In situ identification of the synthrophic protein fermentative Coprothermobacter spp. involved in the thermophilic anaerobic digestion process.

Science.gov (United States)

Gagliano, Maria Cristina; Braguglia, Camilla Maria; Rossetti, Simona

2014-09-01

Thermophilic bacteria have recently attracted great attention because of their potential application in improving different biochemical processes such as anaerobic digestion of various substrates, wastewater treatment or hydrogen production. In this study we report on the design of a specific 16S rRNA-targeted oligonucleotide probe for detecting members of Coprothermobacter genus characterized by a strong protease activity to degrade proteins and peptides. The newly designed CTH485 probe and helper probes hCTH429 and hCTH439 were optimized for use in fluorescence in situ hybridization (FISH) on thermophilic anaerobic sludge samples. In situ probing revealed that thermo-adaptive mechanisms shaping the 16S rRNA gene may affect the identification of thermophilic microorganisms. The novel developed FISH probe extends the possibility to study the widespread thermophilic syntrophic interaction of Coprothermobacter spp. with hydrogenotrophic methanogenic archaea, whose establishment is a great benefit for the whole anaerobic system. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Isolation of thermophilic Desulfotomaculum strains with methanol and sulphite from solfataric mud pools, and characterization of Desulfotomaculum solfataricum

NARCIS (Netherlands)

Goorissen, H.P.; Stams, A.J.M.; Hansen, T.A.

2003-01-01

Four strains of thermophilic, endospore-forming, sulfate-reducing bacteria were enriched and isolated from hot solfataric fields in the Krafla area of north-east Iceland, using methanol and sulfite as substrates. Morphologically, these strains resembled thermophilic Desulfotomaculum species. The

Energetic and hydrogen limitations of thermophilic and hyperthermophilic methanogens

Science.gov (United States)

Stewart, L. C.; Holden, J. F.

2013-12-01

Deep-sea hydrothermal vents are a unique ecosystem, based ultimately not on photosynthesis but chemosynthetic primary production. This makes them an excellent analog environment for the early Earth, and for potential extraterrestrial habitable environments, such as those on Mars and Europa. The habitability of given vent systems for chemoautotrophic prokaryotes can be modeled energetically by estimating the available Gibbs energy for specific modes of chemoautotrophy, using geochemical data and mixing models for hydrothermal fluids and seawater (McCollom and Shock, 1997). However, modeling to date has largely not taken into account variation in organisms' energy demands in these environments. Controls on maintenance energies are widely assumed to be temperature-dependent, rising with increasing temperature optima (Tijhuis et al., 1993), and species-independent. The impacts of other environmental stressors and particular energy-gathering strategies on maintenance energies have not been investigated. We have undertaken culture-based studies of growth and maintenance energies in thermophilic and hyperthermophilic methanogenic (hydrogenotrophic) archaea from deep-sea hydrothermal vents to investigate potential controls on energy demands in hydrothermal vent microbes, and to quantify their growth and maintenance energies for future bioenergetic modeling. We have investigated trends in their growth energies over their full temperature range and a range of nitrogen concentrations, and in their maintenance energies at different hydrogen concentrations. Growth energies in these organisms appear to rise with temperature, but do not vary between hyperthermophilic and thermophilic methanogens. Nitrogen availability at tested levels (40μM - 9.4 mM) does not appear to affect growth energies in all but one tested organism. In continuous chemostat culture, specific methane production varied with hydrogen availability but was similar between a thermophilic and a hyperthermophilic

Plastic degradation by thermophilic Bacillus sp. BCBT21 isolated from composting agricultural residual in Vietnam

Science.gov (United States)

Dang, Thi Cam Ha; Thang Nguyen, Dang; Thai, Hoang; Chinh Nguyen, Thuy; Thu Hien Tran, Thi; Le, Viet Hung; Huynh Nguyen, Van; Bach Tran, Xuan; Phuong Thao Pham, Thi; Giang Nguyen, Truong; Nguyen, Quang Trung

2018-03-01

Three different kinds of plastic bags HL, VHL, and VN1 with different chemical nature were degraded by a novel thermophilic bacterial strain isolated from composting agricultural residual in Vietnam in shaking liquid medium at 55 °C after 30 d. The new strain was classified in the Bacillus genus by morphological property and sequence of partial 16Sr RNA coding gene and named as Bacillus sp. BCBT21. This strain could produce extracellular hydrolase enzymes including lipase, CMCase, xylanase, chitinase, and protease with different level of activity in the same media. After a 30-d treatment at 55 °C with Bacillus sp. BCBT21, all characteristics including properties and morphology of treated plastic bags had been significantly changed. The weight loss, structure and surface morphology of these bags as well as the change in the average molecular weight of VHL bag were detected. Especially, the average molecular weight of VHL bag was significantly reduced from 205 000 to 116 760. New metabolites from the treated bags indicated biodegradation occurring with the different pathways. This finding suggests that there is high potential to develop an effective integrated method for plastic bags degradation by a combination of extracellular enzymes from bacteria and fungi existing in the composting process.

Responses of mycorrhizal fungi and other rootassociated fungi to climate change

DEFF Research Database (Denmark)

Merrild, Marie Porret

Climate change is expected to affect many terrestrial ecosystem processes. Mycorrhizal fungi are important to soil carbon (C) and nutrient cycling thus changes in abundance of mycorrhizal fungi could alter ecosystem functioning. The aim of the present thesis was therefore to investigate responses...... of mycorrhizal fungi to climate change in a seasonal and long-term perspective. Effects of elevated CO2 (510 ppm), night-time warming and extended summer drought were investigated in the long-term field experiment CLIMAITE located in a Danish semi-natural heathland. Mycorrhizal colonization was investigated...... levels. Colonization by arbuscular mycorrhizal (AM) fungi increased under elevated CO2 and warming in spring while ericoid mycorrhiza (ErM) colonisation decreased in response to drought and warming. Increased AM colonization correlated with higher phosphorus and nitrogen root pools. Dark septate...

Chlorophyll a with a farnesyl tail in thermophilic cyanobacteria.

Science.gov (United States)

Wiwczar, Jessica M; LaFountain, Amy M; Wang, Jimin; Frank, Harry A; Brudvig, Gary W

2017-11-01

Photosystem II (PSII) of oxygenic photosynthetic organisms normally contains exclusively chlorophyll a (Chl a) as its major light-harvesting pigment. Chl a canonically consists of the chlorin headgroup with a 20-carbon, 4-isoprene unit, phytyl tail. We have examined the 1.9 Å crystal structure of PSII from thermophilic cyanobacteria reported by Shen and coworkers in 2012 (PDB accession of 3ARC/3WU2). A newly refined electron density map from this structure, presented here, reveals that some assignments of the cofactors may be different from those modeled in the 3ARC/3WU2 structure, including a specific Chl a that appears to have a truncated tail by one isoprene unit. We provide experimental evidence using high-performance liquid chromatography and mass spectrometry for a small population of Chl a esterified to a 15-carbon farnesyl tail in PSII of thermophilic cyanobacteria.

Thermophilic anaerobes in arctic marine sediments induced to mineralize complex organic matter at high temperature

DEFF Research Database (Denmark)

Hubert, Casey; Arnosti, Carol; Brüchert, Volker

2010-01-01

Marine sediments harbour diverse populations of dormant thermophilic bacterial spores that become active in sediment incubation experiments at much higher than in situ temperature. This response was investigated in the presence of natural complex organic matter in sediments of two Arctic fjords......, as well as with the addition of freeze-dried Spirulina or individual high-molecular-weight polysaccharides. During 50°C incubation experiments, Arctic thermophiles catalysed extensive mineralization of the organic matter via extracellular enzymatic hydrolysis, fermentation and sulfate reduction. This high...... reactivity determined the extent of the thermophilic response. Fjord sediments with higher in situ SRR also supported higher SRR at 50°C. Amendment with Spirulina significantly increased volatile fatty acids production and SRR relative to unamended sediment in 50°C incubations. Spirulina amendment also...

Sexual crossing of thermophilic fungus Myceliophthora heterothallica improved enzymatic degradation of sugar beet pulp.

Science.gov (United States)

Aguilar-Pontes, Maria Victoria; Zhou, Miaomiao; van der Horst, Sjors; Theelen, Bart; de Vries, Ronald P; van den Brink, Joost

2016-01-01

Enzymatic degradation of plant biomass requires a complex mixture of many different enzymes. Like most fungi, thermophilic Myceliophthora species therefore have a large set of enzymes targeting different linkages in plant polysaccharides. The majority of these enzymes have not been functionally characterized, and their role in plant biomass degradation is unknown. The biotechnological challenge is to select the right set of enzymes to efficiently degrade a particular biomass. This study describes a strategy using sexual crossing and screening with the thermophilic fungus Myceliophthora heterothallica to identify specific enzymes associated with improved sugar beet pulp saccharification. Two genetically diverse M. heterothallica strains CBS 203.75 and CBS 663.74 were used to generate progenies with improved growth on sugar beet pulp. One progeny, named SBP.F1.2.11, had a different genetic pattern from the parental strains and had improved saccharification activity after the growth on 3 % sugar beet pulp. The improved SBP saccharification was not explained by altered activities of the major (hemi-)cellulases. Exo-proteome analysis of progeny and parental strains after 7-day growth on sugar beet pulp showed that only 17 of the 133 secreted CAZy enzymes were more abundant in progeny SBP.F1.2.11. Particularly one enzyme belonging to the carbohydrate esterase family 5 (CE5) was more abundant in SBP.F1.2.11. This CE5-CBM1 enzyme, named as Axe1, was phylogenetically related to acetyl xylan esterases. Biochemical characterization of Axe1 confirmed de-acetylation activity with optimal activities at 75-85 °C and pH 5.5-6.0. Supplementing Axe1 to CBS 203.75 enzyme set improved release of xylose and glucose from sugar beet pulp. This study identified beneficial enzymes for sugar beet pulp saccharification by selecting progeny with improved growth on this particular substrate. Saccharification of sugar beet pulp was improved by supplementing enzyme mixtures with a previously

Production of Bioethanol From Lignocellulosic Biomass Using Thermophilic Anaerobic Bacteria

DEFF Research Database (Denmark)

Georgieva, Tania I.

2006-01-01

and xylose and to tolerate the inhibitory compounds present in lignocellulosic hydrolysates is therefore apparent. Several thermophilic anaerobic xylan degrading bacteria from our culture collection (EMB group at BioCentrum-DTU) have been screened for a potential ethanol producer from hemicellulose...... hydrolysates, and out of the screening test, one particular strain (A10) was selected for the best performance. The strain was morphologically and physiologically characterized as Thermoanaerobacter mathranii strain A10. Unlike other thermophilic anaerobic bacteria, the wild-type strain Thermoanaerobacter...... Thermoanaerobacter BG1L1 was further studied. The experiments were carried out in a continuous immobilized reactor system (a fluidized bed reactor), which is likely to be the process design configuration for xylose fermentation in a Danish biorefinery concept for production of fuel ethanol. The immobilization...

One-pot strategy for on-site enzyme production, biomass hydrolysis, and ethanol production using the whole solid-state fermentation medium of mixed filamentous fungi.

Science.gov (United States)

Maehara, Larissa; Pereira, Sandra C; Silva, Adilson J; Farinas, Cristiane S

2018-02-01

The efficient use of renewable lignocellulosic feedstocks to obtain biofuels and other bioproducts is a key requirement for a sustainable biobased economy. This requires novel and effective strategies to reduce the cost contribution of the cellulolytic enzymatic cocktails needed to convert the carbohydrates into simple sugars, in order to make large-scale commercial processes economically competitive. Here, we propose the use of the whole solid-state fermentation (SSF) medium of mixed filamentous fungi as an integrated one-pot strategy for on-site enzyme production, biomass hydrolysis, and ethanol production. Ten different individual and mixed cultivations of commonly used industrial filamentous fungi (Aspergillus niger, Aspergillus oryzae, Trichoderma harzianum, and Trichoderma reesei) were performed under SSF and the whole media (without the extraction step) were used in the hydrolysis of pretreated sugarcane bagasse. The cocultivation of T. reesei with A. oryzae increased the amount of glucose released by around 50%, compared with individual cultivations. The release of glucose and reducing sugars achieved using the whole SSF medium was around 3-fold higher than obtained with the enzyme extract. The addition of soybean protein (0.5% w/w) during the hydrolysis reaction further significantly improved the saccharification performance by blocking the lignin and avoiding unproductive adsorption of enzymes. The results of the alcoholic fermentation validated the overall integrated process, with a volumetric ethanol productivity of 4.77 g/L.h, representing 83.5% of the theoretical yield. These findings demonstrate the feasibility of the proposed one-pot integrated strategy using the whole SSF medium of mixed filamentous fungi for on-site enzymes production, biomass hydrolysis, and ethanol production. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018. © 2018 American Institute of Chemical Engineers.

Investigation of the thermophilic mechanism in the genus Porphyrobacter by comparative genomic analysis.

Science.gov (United States)

Xu, Lin; Wu, Yue-Hong; Zhou, Peng; Cheng, Hong; Liu, Qian; Xu, Xue-Wei

2018-05-23

Type strains of the genus Porphyrobacter belonging to the family Erythrobacteraceae and the class Alphaproteobacteria have been isolated from various environments, such as swimming pools, lake water and hot springs. P. cryptus DSM 12079 T and P. tepidarius DSM 10594 T out of all Erythrobacteraceae type strains, are two type strains that have been isolated from geothermal environments. Next-generation sequencing (NGS) technology offers a convenient approach for detecting situational types based on protein sequence differences between thermophiles and mesophiles; amino acid substitutions can lead to protein structural changes, improving the thermal stabilities of proteins. Comparative genomic studies have revealed that different thermal types exist in different taxa, and few studies have been focused on the class Alphaproteobacteria, especially the family Erythrobacteraceae. In this study, eight genomes of Porphyrobacter strains were compared to elucidate how Porphyrobacter thermophiles developed mechanisms to adapt to thermal environments. P. cryptus DSM 12079 T grew optimally at 50 °C, which was higher than the optimal growth temperature of other Porphyrobacter type strains. Phylogenomic analysis of the genus Porphyrobacter revealed that P. cryptus DSM 12079 T formed a distinct and independent clade. Comparative genomic studies uncovered that 1405 single-copy genes were shared by Porphyrobacter type strains. Alignments of single-copy proteins showed that various types of amino acid substitutions existed between P. cryptus DSM 12079 T and the other Porphyrobacter strains. The primary substitution types were changes from glycine/serine to alanine. P. cryptus DSM 12079 T was the sole thermophile within the genus Porphyrobacter. Phylogenomic analysis and amino acid frequencies indicated that amino acid substitutions might play an important role in the thermophily of P. cryptus DSM 12079 T . Bioinformatic analysis revealed that major amino acid substitutional types

Mesophilic and thermophilic anaerobic biodegradability of water hyacinth pre-treated at 80 degrees C.

Science.gov (United States)

Ferrer, Ivet; Palatsi, Jordi; Campos, Elena; Flotats, Xavier

2010-10-01

Water hyacinth (Eichornia crassipes) is a fast growing aquatic plant which causes environmental problems in continental water bodies. Harvesting and handling this plant becomes an issue, and focus has been put on the research of treatment alternatives. Amongst others, energy production through biomethanation has been proposed. The aim of this study was to assess the anaerobic biodegradability of water hyacinth under mesophilic and thermophilic conditions. The effect of a thermal sludge pre-treatment at 80 degrees C was also evaluated. To this end, anaerobic biodegradability tests were carried out at 35 degrees C and 55 degrees C, with raw and pre-treated water hyacinth. According to the results, the thermal pre-treatment enhanced the solubilisation of water hyacinth (i.e. increase in the soluble to total chemical oxygen demand (COD)) from 4% to 12% after 30 min. However, no significant effect was observed on the methane yields (150-190 L CH(4)/kg volatile solids). Initial methane production rates for thermophilic treatments were two fold those of mesophilic ones (6-6.5L vs. 3-3.5 L CH(4)/kg COD x day). Thus, higher methane production rates might be expected from thermophilic reactors working at short retention times. The study of longer low temperature pre-treatments or pre-treatments at elevated temperatures coupled to thermophilic reactors should be considered in the future. (c) 2009 Elsevier Ltd. All rights reserved.

Mesophilic and thermophilic anaerobic biodegradability of water hyacinth pre-treated at 80 oC

International Nuclear Information System (INIS)

Ferrer, Ivet; Palatsi, Jordi; Campos, Elena; Flotats, Xavier

2010-01-01

Water hyacinth (Eichornia crassipes) is a fast growing aquatic plant which causes environmental problems in continental water bodies. Harvesting and handling this plant becomes an issue, and focus has been put on the research of treatment alternatives. Amongst others, energy production through biomethanation has been proposed. The aim of this study was to assess the anaerobic biodegradability of water hyacinth under mesophilic and thermophilic conditions. The effect of a thermal sludge pre-treatment at 80 o C was also evaluated. To this end, anaerobic biodegradability tests were carried out at 35 o C and 55 o C, with raw and pre-treated water hyacinth. According to the results, the thermal pre-treatment enhanced the solubilisation of water hyacinth (i.e. increase in the soluble to total chemical oxygen demand (COD)) from 4% to 12% after 30 min. However, no significant effect was observed on the methane yields (150-190 L CH 4 /kg volatile solids). Initial methane production rates for thermophilic treatments were two fold those of mesophilic ones (6-6.5 L vs. 3-3.5 L CH 4 /kg COD.day). Thus, higher methane production rates might be expected from thermophilic reactors working at short retention times. The study of longer low temperature pre-treatments or pre-treatments at elevated temperatures coupled to thermophilic reactors should be considered in the future.

Changes of resistome, mobilome and potential hosts of antibiotic resistance genes during the transformation of anaerobic digestion from mesophilic to thermophilic.

Science.gov (United States)

Tian, Zhe; Zhang, Yu; Yu, Bo; Yang, Min

2016-07-01

This study aimed to reveal how antibiotic resistance genes (ARGs) and their horizontal and vertical transfer-related items (mobilome and bacterial hosts) respond to the transformation of anaerobic digestion (AD) from mesophilic to thermophilic using one-step temperature increase. The resistomes and mobilomes of mesophilic and thermophilic sludge were investigated using metagenome sequencing, and the changes in 24 representative ARGs belonging to three categories, class 1 integron and bacterial genera during the transition period were further followed using quantitative PCR and 454-pyrosequencing. After the temperature increase, resistome abundance in the digested sludge decreased from 125.97 ppm (day 0, mesophilic) to 50.65 ppm (day 57, thermophilic) with the reduction of most ARG types except for the aminoglycoside resistance genes. Thermophilic sludge also had a smaller mobilome, including plasmids, insertion sequences and integrons, than that of mesophilic sludge, suggesting the lower horizontal transfer potential of ARGs under thermophilic conditions. On the other hand, the total abundance of 18 bacterial genera, which were suggested as the possible hosts for 13 ARGs through network analysis, decreased from 23.27% in mesophilic sludge to 11.92% in thermophilic sludge, indicating fewer hosts for the vertical expansion of ARGs after the increase in temperature. These results indicate that the better reduction of resistome abundance by thermophilic AD might be associated with the decrease of both the horizontal and vertical transferability of ARGs. Copyright © 2016 Elsevier Ltd. All rights reserved.

Comparative microbiological-hygienic studies in mesophilic and thermophilic fouling of sewage sludge

Energy Technology Data Exchange (ETDEWEB)

Pohlig-Schmitt, M.; Philipp, W.; Wekerle, J.; Strauch, D.

Investigations concerning the inactivation of microbial pathogens (bacteria, viruses and parasites) during anaerobic, alkaline dignestion of sludge are described. A thermophilic (54/sup 0/C) and a mesophilic (34/sup 0/C) operated biogas model plant were compared from the point of view of hygiene. Is was found that in the thermophilic process Salmonella senftenberg survived 13,5 h, Streptococcus faecium 55 h, Streptococcus faecalis 42 h and Klebsiella pneumoniae 0,5 h. Within 30 min eggs of Ascaris suum lost their infectivity Bovine Parvovirus was inactivated after 1 d to 2 d treatment. Survival times under mesophilic conditions of 13 d for Salmonella senftenberg and more than 8 mouth for Streptococcus faecium were found. Poliovirus Type 1 was inactivated in 8 d while Bovine Parvovirus survived no longer than 15 d. The results obtained in the thermophilic process were compared to those after heat treatment of the test microorganisms in ampules exposed in a wather-bath under defined conditions to 54/sup 0/C. It was found, that the bacteria survived only about half the time in this case. Poliovirus Type 1 was inactivated after 0,75 h and Bovine Parvovirus after 7 d exposure. (orig.RB)

Direct ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes

Energy Technology Data Exchange (ETDEWEB)

Yanase, Shuhei; Yamada, Ryosuke; Ogino, Chiaki; Kondo, Akihiko [Kobe Univ. (Japan). Dept. of Chemical Science and Engineering; Hasunuma, Tomohisa; Tanaka, Tsutomu; Fukuda, Hideki [Kobe Univ. (Japan). Organization of Advanced Science and Technology

2010-09-15

To exploit cellulosic materials for fuel ethanol production, a microorganism capable of high temperature and simultaneous saccharification-fermentation has been required. However, a major drawback is the optimum temperature for the saccharification and fermentation. Most ethanol-fermenting microbes have an optimum temperature for ethanol fermentation ranging between 28 C and 37 C, while the activity of cellulolytic enzymes is highest at around 50 C and significantly decreases with a decrease in temperature. Therefore, in the present study, a thermotolerant yeast, Kluyveromyces marxianus, which has high growth and fermentation at elevated temperatures, was used as a producer of ethanol from cellulose. The strain was genetically engineered to display Trichoderma reesei endoglucanase and Aspergillus aculeatus {beta}-glucosidase on the cell surface, which successfully converts a cellulosic {beta}-glucan to ethanol directly at 48 C with a yield of 4.24 g/l from 10 g/l within 12 h. The yield (in grams of ethanol produced per gram of {beta}-glucan consumed) was 0.47 g/g, which corresponds to 92.2% of the theoretical yield. This indicates that high-temperature cellulose fermentation to ethanol can be efficiently accomplished using a recombinant K. marxianus strain displaying thermostable cellulolytic enzymes on the cell surface. (orig.)

Production of α-amylase from some thermophilic Aspergillus species ...

African Journals Online (AJOL)

In this study, thermostable amylase activities of some thermophilic Aspergillus species were evaluated. The suitable medium and microorganisms for α-amylase synthesis were selected. Subsequently, the α-amylase activity of the microorganism was researched. In the measurements made on the 7th day of production on ...

The chemical properties and microbial community characterization of the thermophilic microaerobic pretreatment process.

Science.gov (United States)

Fu, Shan-Fei; He, Shuai; Shi, Xiao-Shuang; Katukuri, Naveen Reddy; Dai, Meng; Guo, Rong-Bo

2015-12-01

Thermophilic microaerobic pretreatment (TMP) was recently reported as an efficient pretreatment method of anaerobic digestion (AD). In this study, the chemical properties and microbial community were characterized to reveal how TMP working. Compared with thermophilic treatment under anaerobic condition (TMP0), cellulase activity obviously improved under microaerobic condition (TMP1), which was 10.9-49.0% higher than that of TMP0. Reducing sugar, SCOD and VFAs concentrations of TMP1 were 2.6-8.9%, 1.8-4.8% and 13.8-24% higher than those of TMP0, respectively. TMP gave obvious rise to phylum Firmicutes, which associated with extracellular enzymes production. The proportion of class Bacilli (belongs to phylum Firmicutes and mainly acts during hydrolysis) in TMP1 was 124.89% higher than that of TMP0, which reflected the greater hydrolytic ability under microaerobic condition. The improved abundance of phylum Firmicutes (especially class Bacilli, order Bacillales) under microaerobic condition could be the fundamental reason for the improved AD performance of thermophilic microaerobic pretreated corn straw. Copyright © 2015 Elsevier Ltd. All rights reserved.

The Feasibility of Thermophilic Caldimonas manganoxidans as a Platform for Efficient PHB Production.

Science.gov (United States)

Hsiao, Li-Jung; Lin, Ji-Hong; Sankatumvong, Pantitra; Wu, Tzong-Ming; Li, Si-Yu

2016-11-01

Recently, poly(3-hydroxybutyrate) (PHB) has been found in a few thermophilic strains where several advantages can be gained from running fermentation at high temperatures. Caldimonas manganoxidans, a thermophilic gram-negative bacterium, was investigated for the feasibility as a PHB-producing strain. It is suggested that the best fermentation strategy for achieving the highest PHB concentration of 5.4 ± 1.1 g/L (from 20 g/L glucose) in 24 h is to use the fermentation conditions that are favored for the bacterial growth, yet temperature and pH should be chosen at conditions that are favored for the PHB content. Besides, the above fermentation conditions produce PHB that has a high molecular weight of 1274 kDa with a low polydispersity index (PDI) of 1.45, where the highest Mw of PHB of 1399 kDa (PDI of 1.32) is obtained in this study. To the best knowledge of authors, C. manganoxidans has the best PHB productivity among the thermophiles and is comparable to those common PHB-producing mesophiles.

Genes regulated by AoXlnR, the xylanolytic and cellulolytic transcriptional regulator, in Aspergillus oryzae.

Science.gov (United States)

Noguchi, Yuji; Sano, Motoaki; Kanamaru, Kyoko; Ko, Taro; Takeuchi, Michio; Kato, Masashi; Kobayashi, Tetsuo

2009-11-01

XlnR is a Zn(II)2Cys6 transcriptional activator of xylanolytic and cellulolytic genes in Aspergillus. Overexpression of the aoxlnR gene in Aspergillus oryzae (A. oryzae xlnR gene) resulted in elevated xylanolytic and cellulolytic activities in the culture supernatant, in which nearly 40 secreted proteins were detected by two-dimensional electrophoresis. DNA microarray analysis to identify the transcriptional targets of AoXlnR led to the identification of 75 genes that showed more than fivefold increase in their expression in the AoXlnR overproducer than in the disruptant. Of these, 32 genes were predicted to encode a glycoside hydrolase, highlighting the biotechnological importance of AoXlnR in biomass degradation. The 75 genes included the genes previously identified as AoXlnR targets (xynF1, xynF3, xynG2, xylA, celA, celB, celC, and celD). Thirty-six genes were predicted to be extracellular, which was consistent with the number of proteins secreted, and 61 genes possessed putative XlnR-binding sites (5'-GGCTAA-3', 5'-GGCTAG-3', and 5'-GGCTGA-3') in their promoter regions. Functional annotation of the genes revealed that AoXlnR regulated the expression of hydrolytic genes for degradation of beta-1,4-xylan, arabinoxylan, cellulose, and xyloglucan and of catabolic genes for the conversion of D-xylose to xylulose-5-phosphate. In addition, genes encoding glucose-6-phosphate 1-dehydrogenase and L-arabinitol-4- dehydrogenase involved in D-glucose and L-arabinose catabolism also appeared to be targets of AoXlnR.

Protein dynamics and stability: The distribution of atomic fluctuations in thermophilic and mesophilic dihydrofolate reductase derived using elastic incoherent neutron scattering

International Nuclear Information System (INIS)

Meinhold, Lars; Clement, David; Tehei, M.; Daniel, R.M.; Finney, J.L.; Smith, Jeremy C.

2008-01-01

The temperature dependence of the dynamics of mesophilic and thermophilic dihydrofolate reductase is examined using elastic incoherent neutron scattering. It is demonstrated that the distribution of atomic displacement amplitudes can be derived from the elastic scattering data by assuming a (Weibull) functional form that resembles distributions seen in molecular dynamics simulations. The thermophilic enzyme has a significantly broader distribution than its mesophilic counterpart. Furthermore, although the rate of increase with temperature of the atomic mean-square displacements extracted from the dynamic structure factor is found to be comparable for both enzymes, the amplitudes are found to be slightly larger for the thermophilic enzyme. Therefore, these results imply that the thermophilic enzyme is the more flexible of the two

Fungi in space--literature survey on fungi used for space research.

Science.gov (United States)

Kern, V D; Hock, B

1993-09-01

A complete review of the scientific literature on experiments involving fungi in space is presented. This review begins with balloon experiments around 1935 which carried fungal spores, rocket experiments in the 1950's and 60's, satellite and moon expeditions, long-time orbit experiments and Spacelab missions in the 1980's and 90's. All these missions were aimed at examining the influence of cosmic radiation and weightlessness on genetic, physiological, and morphogenetic processes. During the 2nd German Spacelab mission (D-2, April/May 1993), the experiment FUNGI provided the facilities to cultivate higher basidiomycetes over a period of 10 d in orbit, document gravimorphogenesis and chemically fix fruiting bodies under weightlessness for subsequent ultrastructural analysis. This review shows the necessity of space travel for research on the graviperception of higher fungi and demonstrates the novelty of the experiment FUNGI performed within the framework of the D-2 mission.

Gas Fermentation using Thermophilic Moorella Species for production of Biochemicals

DEFF Research Database (Denmark)

Redl, Stephanie Maria Anna

Gas fermentation is a promising technology which gained increasing attention over the last years. In this process, acetogenic bacteria convert gases rich in H2, CO2, and CO, into compounds of higher value. The gas can derive from industrial off-gas or from waste streams via gasification. In the gas...... fermentation processes that are nearly on commercial level, mesophilic acetogens are used to mainly produce ethanol and butanediol. However, thermophilic acetogens, such as Moorella thermoacetica would allow for easy downstream processing when producing volatile products such as acetone. This thesis starts...... with a review of the feedstock potential for gas fermentation and how thermophilic production strains as well as unconventional fermentation processes such as mixotrophy can help to exploit this potential. I analyzed a process with respect to thermodynamic and economic considerations, in which acetone...

A rigidifying salt-bridge favors the activity of thermophilic enzyme at high temperatures at the expense of low-temperature activity.

Science.gov (United States)

Lam, Sonia Y; Yeung, Rachel C Y; Yu, Tsz-Ha; Sze, Kong-Hung; Wong, Kam-Bo

2011-03-01

Thermophilic enzymes are often less active than their mesophilic homologues at low temperatures. One hypothesis to explain this observation is that the extra stabilizing interactions increase the rigidity of thermophilic enzymes and hence reduce their activity. Here we employed a thermophilic acylphosphatase from Pyrococcus horikoshii and its homologous mesophilic acylphosphatase from human as a model to study how local rigidity of an active-site residue affects the enzymatic activity. Acylphosphatases have a unique structural feature that its conserved active-site arginine residue forms a salt-bridge with the C-terminal carboxyl group only in thermophilic acylphosphatases, but not in mesophilic acylphosphatases. We perturbed the local rigidity of this active-site residue by removing the salt-bridge in the thermophilic acylphosphatase and by introducing the salt-bridge in the mesophilic homologue. The mutagenesis design was confirmed by x-ray crystallography. Removing the salt-bridge in the thermophilic enzyme lowered the activation energy that decreased the activation enthalpy and entropy. Conversely, the introduction of the salt-bridge to the mesophilic homologue increased the activation energy and resulted in increases in both activation enthalpy and entropy. Revealed by molecular dynamics simulations, the unrestrained arginine residue can populate more rotamer conformations, and the loss of this conformational freedom upon the formation of transition state justified the observed reduction in activation entropy. Our results support the conclusion that restricting the active-site flexibility entropically favors the enzymatic activity at high temperatures. However, the accompanying enthalpy-entropy compensation leads to a stronger temperature-dependency of the enzymatic activity, which explains the less active nature of the thermophilic enzymes at low temperatures.

A rigidifying salt-bridge favors the activity of thermophilic enzyme at high temperatures at the expense of low-temperature activity.

Directory of Open Access Journals (Sweden)

Sonia Y Lam

2011-03-01

Full Text Available Thermophilic enzymes are often less active than their mesophilic homologues at low temperatures. One hypothesis to explain this observation is that the extra stabilizing interactions increase the rigidity of thermophilic enzymes and hence reduce their activity. Here we employed a thermophilic acylphosphatase from Pyrococcus horikoshii and its homologous mesophilic acylphosphatase from human as a model to study how local rigidity of an active-site residue affects the enzymatic activity.Acylphosphatases have a unique structural feature that its conserved active-site arginine residue forms a salt-bridge with the C-terminal carboxyl group only in thermophilic acylphosphatases, but not in mesophilic acylphosphatases. We perturbed the local rigidity of this active-site residue by removing the salt-bridge in the thermophilic acylphosphatase and by introducing the salt-bridge in the mesophilic homologue. The mutagenesis design was confirmed by x-ray crystallography. Removing the salt-bridge in the thermophilic enzyme lowered the activation energy that decreased the activation enthalpy and entropy. Conversely, the introduction of the salt-bridge to the mesophilic homologue increased the activation energy and resulted in increases in both activation enthalpy and entropy. Revealed by molecular dynamics simulations, the unrestrained arginine residue can populate more rotamer conformations, and the loss of this conformational freedom upon the formation of transition state justified the observed reduction in activation entropy.Our results support the conclusion that restricting the active-site flexibility entropically favors the enzymatic activity at high temperatures. However, the accompanying enthalpy-entropy compensation leads to a stronger temperature-dependency of the enzymatic activity, which explains the less active nature of the thermophilic enzymes at low temperatures.

ESTIMATION OF EXTRACELLULAR LIPOLYTIC ENZYME ACTIVITY BY THERMOPHILIC BACILLUS SP. ISOLATED FROM ARID AND SEMI-ARID REGION OF RAJASTHAN, INDIA

Directory of Open Access Journals (Sweden)

Deeksha Gaur

2012-10-01

Full Text Available Thermophilic organisms can be defined as, micro-organisms which are adapted to survive at high temperatures. The enzymes secreted by thermophilic bacteria are capable of catalyzing biochemical reactions at high temperatures. Thermophilic bacteria are able to produce thermostable lipolytic enzymes (capable of degradation of lipid at temperatures higher than mesophilic bacteria. Therefore, the isolation of thermophilic bacteria from natural sources and their identification are quite beneficial in terms of discovering thermostable lipase enzymes. Due to great temperature fluctuation in hot arid and semi-arid region of Rajasthan, this area could serve as a good source for new thermophilic lipase producing bacteria with novel industrially important properties. The main objective of this research is the isolation and estimation of industrially important thermophilic lipase enzyme produced by thermophilic bacteria, isolated from arid and semi-arid region of Rajasthan. For this research purpose soil samples were collected from Churu, Sikar and Jhunjunu regions of Rajasthan. Total 16 bacterial strains were isolated and among only 2 thermostable lipolytic enzyme producing bacteria were charcterized. The thermostable lipolytic enzyme was estimated by qualitative and quantitative experiments. The isolates were identified as Bacillus sp. by microscopic, biochemical and molecular characterization. The optimum enzyme activity was observed at pH 8, temperature 60°C and 6% salt concentrations at 24 hrs time duration. Lipolytic enzyme find useful in a variety of biotechnological fields such as food and dairy (cheese ripening, flavour development, detergent, pharmaceutical (naproxen, ibuprofen, agrochemical (insecticide, pesticide and oleochemical (fat and oil hydrolysis, biosurfactant synthesis industries. Lipolytic enzyme can be further used in many newer areas where they can serve as potential biocatalysts.

DHAP-dependent aldolases from (hyper)thermophiles: biochemistry and applications

NARCIS (Netherlands)

Falcicchio, P.; Wolterink-van Loo, S.; Franssen, M.C.R.; Oost, van der J.

2014-01-01

Generating new carbon-carbon (C-C) bonds in an enantioselective way is one of the big challenges in organic synthesis. Aldolases are a natural tool for stereoselective C-C bond formation in a green and sustainable way. This review will focus on thermophilic aldolases in general and on

Azo dye reduction by mesophilic and thermophilic anaerobic consortia

NARCIS (Netherlands)

Santos, dos A.B.; Madrid, de M.P.; Stams, A.J.M.; Lier, van J.B.; Cervantes, F.J.

2005-01-01

The reduction of the azo dye model compounds Reactive Red 2 (RR2) and Reactive Orange 14 (RO14) by mesophilic (30 C) and thermophilic (55 C) anaerobic consortia was studied in batch assays. The contribution of fermentative and methanogenic microorganisms in both temperatures was evaluated in the

Effect of NaCl on thermophilic (55°C) methanol degradation in sulfate reducing granular sludge reactors

NARCIS (Netherlands)

Vallero, M.V.G.; Hulshoff Pol, L.W.; Lettinga, G.; Lens, P.N.L.

2003-01-01

The effect of NaCl on thermophilic (55degreesC) methanol conversion in the presence of excess of sulfate (COD/SO42-=0.5) was investigated in two 6.5L lab-scale upflow anaerobic sludge bed reactors inoculated with granular sludge previously not adapted to NaCl
The effect of NaCl on thermophilic

Formation and release of cellulolytic enzymes during growth of Trichoderma reesei on cellobiose and glycerol

Energy Technology Data Exchange (ETDEWEB)

Vaheri, M.P.; Vaheri, M.E.O.; Kaupinen, V.S.

1979-01-01

Production and release of cellulolytic enzymes by T. reesei QM 9414 were studied under induced and non-induced conditions and glycerol, respectively, as the only C source. There was a base level of cell debris-bound hydrolytic activity against filter paper and p-nitrophenyl glycoside even in T. reesei grown non-induced on glycerol. T. reesei grown on cellobiose was induced to produce large amounts of extracellular filter paper- and CMC-hydrolyzing enzymes, which were actively released even in the early stages of cultivation. Beta-Glucosidase was mainly detected in the cell debris and was not released unless the cells were autolyzing.

Thermophilic anaerobic oxidation of methane by marine microbial consortia.

Science.gov (United States)

Holler, Thomas; Widdel, Friedrich; Knittel, Katrin; Amann, Rudolf; Kellermann, Matthias Y; Hinrichs, Kai-Uwe; Teske, Andreas; Boetius, Antje; Wegener, Gunter

2011-12-01

The anaerobic oxidation of methane (AOM) with sulfate controls the emission of the greenhouse gas methane from the ocean floor. AOM is performed by microbial consortia of archaea (ANME) associated with partners related to sulfate-reducing bacteria. In vitro enrichments of AOM were so far only successful at temperatures ≤25 °C; however, energy gain for growth by AOM with sulfate is in principle also possible at higher temperatures. Sequences of 16S rRNA genes and core lipids characteristic for ANME as well as hints of in situ AOM activity were indeed reported for geothermally heated marine environments, yet no direct evidence for thermophilic growth of marine ANME consortia was obtained to date. To study possible thermophilic AOM, we investigated hydrothermally influenced sediment from the Guaymas Basin. In vitro incubations showed activity of sulfate-dependent methane oxidation between 5 and 70 °C with an apparent optimum between 45 and 60 °C. AOM was absent at temperatures ≥75 °C. Long-term enrichment of AOM was fastest at 50 °C, yielding a 13-fold increase of methane-dependent sulfate reduction within 250 days, equivalent to an apparent doubling time of 68 days. The enrichments were dominated by novel ANME-1 consortia, mostly associated with bacterial partners of the deltaproteobacterial HotSeep-1 cluster, a deeply branching phylogenetic group previously found in a butane-amended 60 °C-enrichment culture of Guaymas sediments. The closest relatives (Desulfurella spp.; Hippea maritima) are moderately thermophilic sulfur reducers. Results indicate that AOM and ANME archaea could be of biogeochemical relevance not only in cold to moderate but also in hot marine habitats.

Thermophilic archaeal enzymes and applications in biocatalysis.

Science.gov (United States)

Littlechild, Jennifer A

2011-01-01

Thermophilic enzymes have advantages for their use in commercial applications and particularly for the production of chiral compounds to produce optically pure pharmaceuticals. They can be used as biocatalysts in the application of 'green chemistry'. The thermophilic archaea contain enzymes that have already been used in commercial applications such as the L-aminoacylase from Thermococcus litoralis for the resolution of amino acids and amino acid analogues. This enzyme differs from bacterial L-aminoacylases and has similarities to carboxypeptidases from other archaeal species. An amidase/γ-lactamase from Sulfolobus solfataricus has been used for the production of optically pure γ-lactam, the building block for antiviral carbocyclic nucleotides. This enzyme has similarities to the bacterial signature amidase family. An alcohol dehydrogenase from Aeropyrum pernix has been used for the production of optically pure alcohols and is related to the zinc-containing eukaryotic alcohol dehydrogenases. A transaminase and a dehalogenase from Sulfolobus species have also been studied. The archaeal transaminase is found in a pathway for serine synthesis which is found only in eukaryotes and not in bacteria. It can be used for the asymmetric synthesis of homochiral amines of high enantioselective purity. The L-2-haloacid dehalogenase has applications both in biocatalysis and in bioremediation. All of these enzymes have increased thermostability over their mesophilic counterparts.

Philatelic Mycology: Families of Fungi

NARCIS (Netherlands)

Marasas, W.F.O.; Marasas, H.M.; Wingfield, M.J.; Crous, P.W.

2014-01-01

Philately, the study of postage stamps, and mycology, the study of fungi, are seldom connected by those that practice these very different activities. When associated, philatelic mycology would be considered as the study of fungi on stamps. The Fungi touch every aspect of our daily lives, most

Comparison of multi-enzyme and thermophilic bacteria on the hydrolysis of mariculture organic waste (MOW).

Science.gov (United States)

Guo, Liang; Sun, Mei; Zong, Yan; Zhao, Yangguo; Gao, Mengchun; She, Zonglian

2016-01-01

Mariculture organic waste (MOW) is rich in organic matter, which is a potential energy resource for anaerobic digestion. In order to enhance the anaerobic fermentation, the MOW was hydrolyzed by multi-enzyme and thermophilic bacteria. It was advantageous for soluble chemical oxygen demand (SCOD) release at MOW concentrations of 6 and 10 g/L with multi-enzyme and thermophilic bacteria pretreatments. For multi-enzyme, the hydrolysis was not obvious at substrate concentrations of 1 and 3 g/L, and the protein and carbohydrate increased with hydrolysis time at substrate concentrations of 6 and 10 g/L. For thermophilic bacteria, the carbohydrate was first released at 2-4 h and then consumed, and the protein increased with hydrolysis time. The optimal enzyme hydrolysis for MOW was determined by measuring the changes of SCOD, protein, carbohydrate, ammonia and total phosphorus, and comparing with acid and alkaline pretreatments.

Application of ATR-FTIR Spectroscopy to Compare the Cell Materials of Wood Decay Fungi with Wood Mould Fungi

Directory of Open Access Journals (Sweden)

Barun Shankar Gupta

2015-01-01

Full Text Available Wood fungi create vast damage among standing trees and all types of wood materials. The objectives of this study are to (a characterize the cell materials of two major wood decay fungi (Basidiomycota, namely, Trametes versicolor and Postia placenta, and (b compare the cell materials of decay fungi with four wood mould fungi (Ascomycota, namely, Aureobasidium pullulans, Alternaria alternata, Cladosporium cladosporioides, and Ulocladium atrum. Fourier transform infrared (FTIR spectroscopy is used to characterize the microbial cellular materials. The results showed that the IR bands for the fatty acid at ∼2900 cm−1 were different for the two-decay-fungi genre. Postia placenta shows more absorbance peaks at the fatty acid region. Band ratio indices for amide I and amide II from protein amino acids were higher for the mould fungi (Ascomycota than the decay fungi (Basidiomycota. Similarly, the band ratio index calculated for the protein end methyl group was found to be higher for the mould fungi than the decay fungi. Mould fungi along with the decay fungi demonstrated a positive correlation (R2=0.75 between amide I and amide II indices. The three-component multivariate, principal component analysis showed a strong correlation of amide and protein band indices.

TtMCO: A highly thermostable laccase-like multicopper oxidase from the thermophilic Thermobaculum terrenum

DEFF Research Database (Denmark)

Brander, Søren; Mikkelsen, Jørn Dalgaard; Kepp, Kasper Planeta

2015-01-01

This paper reports the identification, heterologous expression in Escherichia coli and characterization of TtMCO from the thermophilic bacterium Thermobaculum terrenum, the first laccase-like multi-copper oxidase (LMCO) from the distinct Phylum Chloroflexi. TtMCO has only 39% identity to its...... closest characterized homologue, CotA from Bacillus subtilis, but sequence and spectrophotometry confirmed copper coordination similar to that of LMCOs. TtMCO is extremely thermophilic with a half-time of inactivation of 2.24 days at 70 degrees C and 350 min at 80°C and pH 7, consistent...

Development of a continuous bioconversion system using a thermophilic whole-cell biocatalyst.

Science.gov (United States)

Ninh, Pham Huynh; Honda, Kohsuke; Yokohigashi, Yukako; Okano, Kenji; Omasa, Takeshi; Ohtake, Hisao

2013-03-01

The heat treatment of recombinant mesophilic cells having heterologous thermophilic enzymes results in the denaturation of indigenous mesophilic enzymes and the elimination of undesired side reactions; therefore, highly selective whole-cell catalysts comparable to purified enzymes can be readily prepared. However, the thermolysis of host cells leads to the heat-induced leakage of thermophilic enzymes, which are produced as soluble proteins, limiting the exploitation of their excellent stability in repeated and continuous reactions. In this study, Escherichia coli cells having the thermophilic fumarase from Thermus thermophilus (TtFTA) were treated with glutaraldehyde to prevent the heat-induced leakage of the enzyme, and the resulting cells were used as a whole-cell catalyst in repeated and continuous reactions. Interestingly, although electron microscopic observations revealed that the cellular structure of glutaraldehyde-treated E. coli was not apparently changed by the heat treatment, the membrane permeability of the heated cells to relatively small molecules (up to at least 3 kDa) was significantly improved. By applying the glutaraldehyde-treated E. coli having TtFTA to a continuous reactor equipped with a cell-separation membrane filter, the enzymatic hydration of fumarate to malate could be operated for more than 600 min with a molar conversion yield of 60% or higher.

Thermophilic anaerobic co-digestion of poultry litter and thin stillage.

Science.gov (United States)

Sharma, Deepak; Espinosa-Solares, Teodoro; Huber, David H

2013-05-01

The purpose of this study was to test whether the performance of a thermophilic CSTR digester that has been stabilized on poultry litter will be enhanced or diminished by the addition of thin stillage as co-substrate. Replicate laboratory digesters, derived from a stable pilot-scale digester, were operated with increasing ratios (w/w) of thin stillage/poultry litter feedstock. After a period of adaptation to 20% and 40% thin stillage, digester performance showed increases in biogas, percent methane and COD removal, as well as a decrease in volatile acids. Peak performance occurred with 60% thin stillage. However, 80% thin stillage caused significant reduction of performance, including declines of methanogenic activity and COD removal. In conclusion, supplementing the thermophilic digestion of poultry litter with thin stillage improved the bioenergy (methane) output, but thin stillage became inhibitory at high concentrations. Copyright © 2013 Elsevier Ltd. All rights reserved.

Biotechnology of marine fungi

Digital Repository Service at National Institute of Oceanography (India)

Damare, S.R.; Singh, P.; Raghukumar, S.

Filamentous fungi are the most widely used eukaryotes in industrial and pharmaceutical applications. Their biotechnological uses include the production of enzymes, vitamins, polysaccharides, pigments, lipids and others. Marine fungi are a still...

Isolation of thermophilic Desulfotomaculum strains with methanol and sulfite from solfataric mud pools, and characterization of Desulfotomaculum solfataficum sp nov

NARCIS (Netherlands)

Goorissen, H.P.; Boschker, H.T.S.; Stams, A.J.M.; Hansen, T.A.

2003-01-01

Four strains of thermophilic, endospore-forming, sulfate-reducing bacteria were enriched and isolated from hot solfataric fields in the Krafla area of north-east Iceland, using methanol and sulfite as substrates. Morphologically, these strains resembled thermophilic Desulfotomaculum species. The

Isolation of thermophilic Desulfotomaculum strains with methanol and sulfite from solfataric mud pools, and characterization of Desulfotomaculum solfataficum sp nov

NARCIS (Netherlands)

Goorissen, HP; Boschker, HTS; Stams, AJM; Hansen, TA

Four strains of thermophilic, endospore-forming, sulfate-reducing bacteria were enriched and isolated from hot solfataric fields in the Krafla area of north-east Iceland, using methanol and sulfite as substrates. Morphologically, these strains resembled thermophilic Desulfotomaculum species. The

Isolation of soil thermophilic strains of actinomycetes for the ...

African Journals Online (AJOL)

use

2011-12-05

Dec 5, 2011 ... to high fructose (Pandey et al., 2000; Asgher et al., 2007). *Corresponding ... can be increased by pH, temperature or substrates. ... The following media were used for isolating thermophilic strains of ... To observe the effect of different culture conditions on α-amylase .... Influence of pH on the inactivation of.

Thermophilic and alkalophilic xylanases from several Dictyoglomus isolates

Energy Technology Data Exchange (ETDEWEB)

Mathrani, I M; Ahring, B K [Technical Univ. of Denmark, Lyngby (Denmark). Anaerobic Microbiology/Biotechnology Group

1992-10-01

Supernatant xylanases from three thermophilic and strictly anaerobic Dictyoglomus strains isolated from very different environments were examined: The type species, D. thermophilum[sup T], from a hot-spring in Japan; strain B1, a recently described strictly xylanutilizing Dictyoglomus from a paper-pulp factory in Finland; and strain B4a, isolated from a thermal pool on Iceland. The highest enzymatic activity observed from batch-culture supernatant with 4 g l[sup -1] of beech xylan as growth substrate was 3.8x10[sup -6] kat l[sup -1]. The K[sub m] for the xylanases of strain B1 was 4.7 g beech xylan l[sup -1]. The xylanases of all the isolates had a broad range of activity with respect to pH, showing good activity from pH 5.5 to near 9.0. The xylanases from the three isolates had a very high temperature optimum of 80deg C, maximum temperature for extended activity between 80 and 90deg C, and a thermal half-life of over 1 h at 90deg C for strain B1. The application of thermophilic alkalophilic xylanases to paper pulping was discussed. (orig.).

Bioleaching of metals from electronic scrap by moderately thermophilic acidophilic bacteria

NARCIS (Netherlands)

Ilyas, Sadia; Anwar, Munir A.; Niazi, Shahida B.; Ghauri, M. Afzal

The present work was aimed at studying the bioleachability of metals from electronic scrap by the selected moderately thermophilic strains of acidophilic chemolithotrophic and acidophilic heterotrophic bacteria. These included Sulfobacillus thermosulfidooxidans and an unidentified acidophilic

Probing the redox metabolism in the strictly anaerobic, extremely thermophilic, hydrogen-producing Caldicellulosiruptor saccharolyticus using amperometry

DEFF Research Database (Denmark)

Kostesha, Natalie; Willquist, Karin; Emnéus, Jenny

2011-01-01

Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellul...... in the intracellular electron flow and to probe redox enzyme properties of a strictly anaerobic thermophile in vivo.......Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellular...... the NADH-dependent lactate dehydrogenase, upon which more NADH was directed to membrane-associated enzymes for ferricyanide reduction, leading to a higher electrochemical signal. The method is noninvasive and the results presented here demonstrate that this method can be used to accurately detect changes...

Author Details

African Journals Online (AJOL)

Johnson, FS. Vol 30, No 1 (2010) - Articles Saccharification of Maize Agrowastes by Cellulolytic Fungi Isolated from Ejura Farms in Ejura, Ghana Abstract PDF. ISSN: 0855-0395. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's · More about AJOL · AJOL's Partners ...

High-solids enrichment of thermophilic microbial communities and their enzymes on bioenergy feedstocks

Energy Technology Data Exchange (ETDEWEB)

Reddy, A. P.; Allgaier, M.; Singer, S.W.; Hazen, T.C.; Simmons, B.A.; Hugenholtz, P.; VanderGheynst, J.S.

2011-04-01

Thermophilic microbial communities that are active in a high-solids environment offer great potential for the discovery of industrially relevant enzymes that efficiently deconstruct bioenergy feedstocks. In this study, finished green waste compost was used as an inoculum source to enrich microbial communities and associated enzymes that hydrolyze cellulose and hemicellulose during thermophilic high-solids fermentation of the bioenergy feedstocks switchgrass and corn stover. Methods involving the disruption of enzyme and plant cell wall polysaccharide interactions were developed to recover xylanase and endoglucanase activity from deconstructed solids. Xylanase and endoglucanase activity increased by more than a factor of 5, upon four successive enrichments on switchgrass. Overall, the changes for switchgrass were more pronounced than for corn stover; solids reduction between the first and second enrichments increased by a factor of four for switchgrass while solids reduction remained relatively constant for corn stover. Amplicon pyrosequencing analysis of small-subunit ribosomal RNA genes recovered from enriched samples indicated rapid changes in the microbial communities between the first and second enrichment with the simplified communities achieved by the third enrichment. The results demonstrate a successful approach for enrichment of unique microbial communities and enzymes active in a thermophilic high-solids environment.

Effect of feeding palm oil by-products based diets on total bacteria, cellulolytic bacteria and methanogenic archaea in the rumen of goats.

Science.gov (United States)

Abubakr, Abdelrahim; Alimon, Abdul Razak; Yaakub, Halimatun; Abdullah, Norhani; Ivan, Michael

2014-01-01

Rumen microorganisms are responsible for digestion and utilization of dietary feeds by host ruminants. Unconventional feed resources could be used as alternatives in tropical areas where feed resources are insufficient in terms of quality and quantity. The objective of the present experiment was to evaluate the effect of diets based on palm oil (PO), decanter cake (DC) or palm kernel cake (PKC) on rumen total bacteria, selected cellulolytic bacteria, and methanogenic archaea. Four diets: control diet (CD), decanter cake diet (DCD), palm kernel cake diet (PKCD) and CD plus 5% PO diet (CPOD) were fed to rumen cannulated goats and rumen samples were collected at the start of the experimental diets (day 0) and on days 4, 6, 8, 12, 18, 24 and 30 post dietary treatments. Feeding DCD and PKCD resulted in significantly higher (Pgoats fed PKCD and CPOD and the trend showed a severe reduction on days 4 and 6 post experimental diets. In conclusion, results indicated that feeding DCD and PKC increased the populations of cellulolytic bacteria and decreased the density of methanogenic archaea in the rumen of goats.

Pressure stabilization is not a general property of thermophilic enzymes: the adenylate kinases of Methanococcus voltae, Methanococcus maripaludis, Methanococcus thermolithotrophicus, and Methanococcus jannaschii.

OpenAIRE

Konisky, J; Michels, P C; Clark, D S

1995-01-01

The application of 50-MPa pressure did not increase the thermostabilities of adenylate kinases purified from four related mesophilic and thermophilic marine methanogens. Thus, while it has been reported that some thermophilic enzymes are stabilized by pressure (D. J. Hei and D. S. Clark, Appl. Environ. Microbiol. 60:932-939, 1994), hyperbaric stabilization is not an intrinsic property of all enzymes from deep-sea thermophiles.

Potential use of thermophilic dark fermentation effluents in photofermentative hydrogen production by Rhodobacter capsulatus

Energy Technology Data Exchange (ETDEWEB)

Ozgura, E.; Afsar, N.; Eroglu, I. [Middle East Technical University, Department of Chemical Engineering, 06531 Ankara (Turkey); De Vrije, T.; Claassen, P.A.M. [Wageningen UR, Agrotechnology and Food Sciences Group, Wageningen UR, P.O. Box 17, 6700 AA Wageningen (Netherlands); Yucel, M.; Gunduz, U. [Middle East Technical University, Department of Biology, 06531 Ankara (Turkey)

2010-12-15

Biological hydrogen production by a sequential operation of dark and photofermentation is a promising route to produce hydrogen. The possibility of using renewable resources, like biomass and agro-industrial wastes, provides a dual effect of sustainability in biohydrogen production and simultaneous waste removal. In this study, photofermentative hydrogen production on effluents of thermophilic dark fermentations on glucose, potato steam peels (PSP) hydrolysate and molasses was investigated in indoor, batch operated bioreactors. An extreme thermophile Caldicellulosiruptor saccharolyticus was used in the dark fermentation step, and Rhodobacter capsulatus (DSM1710) was used in the photofermentation step. Addition of buffer, Fe and Mo to dark fermentor effluents (DFEs) improved the overall efficiency of hydrogen production. The initial acetate concentration in the DFE needed to be adjusted to 30-40 mM by dilution to increase the yield of hydrogen in batch light-supported fermentations. The thermophilic DFEs are suitable for photofermentative hydrogen production, provided that they are supplemented with buffer and nutrients. The overall hydrogen yield of the two-step fermentations was higher than the yield of single step dark fermentations.

Thermophilic Anaerobic Degradation of Butyrate by a Butyrate-Utilizing Bacterium in Coculture and Triculture with Methanogenic Bacteria

OpenAIRE

Ahring, Birgitte K.; Westermann, Peter

1987-01-01

We studied syntrophic butyrate degradation in thermophilic mixed cultures containing a butyrate-degrading bacterium isolated in coculture with Methanobacterium thermoautotrophicum or in triculture with M. thermoautotrophicum and the TAM organism, a thermophilic acetate-utilizing methanogenic bacterium. Butyrate was β-oxidized to acetate with protons as the electron acceptors. Acetate was used concurrently with its production in the triculture. We found a higher butyrate degradation rate in th...

Differences in the catalytic mechanisms of mesophilic and thermophilic indole-3-glycerol phosphate synthase enzymes at their adaptive temperatures

International Nuclear Information System (INIS)

Zaccardi, Margot J.; Mannweiler, Olga; Boehr, David D.

2012-01-01

Highlights: ► Catalytic mechanisms of thermophilic–mesophilic enzymes may differ. ► Product release is rate-determining for thermophilic IGPS at low temperatures. ► But at higher temperatures, proton transfer from the general acid is rate-limiting. ► Rate-determining step is different still for mesophilic IGPS. ► Both chemical and physical steps of catalysis are important for temperature adaptation. -- Abstract: Thermophilic enzymes tend to be less catalytically-active at lower temperatures relative to their mesophilic counterparts, despite having very similar crystal structures. An often cited hypothesis for this general observation is that thermostable enzymes have evolved a more rigid tertiary structure in order to cope with their more extreme, natural environment, but they are also less flexible at lower temperatures, leading to their lower catalytic activity under mesophilic conditions. An alternative hypothesis, however, is that complementary thermophilic–mesophilic enzyme pairs simply operate through different evolutionary-optimized catalytic mechanisms. In this communication, we present evidence that while the steps of the catalytic mechanisms for mesophilic and thermophilic indole-3-glycerol phosphate synthase (IGPS) enzymes are fundamentally similar, the identity of the rate-determining step changes as a function of temperature. Our findings indicate that while product release is rate-determining at 25 °C for thermophilic IGPS, near its adaptive temperature (75 °C), a proton transfer event, involving a general acid, becomes rate-determining. The rate-determining steps for thermophilic and mesophilic IGPS enzymes are also different at their respective, adaptive temperatures with the mesophilic IGPS-catalyzed reaction being rate-limited before irreversible CO 2 release, and the thermophilic IGPS-catalyzed reaction being rate limited afterwards.

Natural substrata for corticioid fungi

Directory of Open Access Journals (Sweden)

Eugene O. Yurchenko

2013-12-01

Full Text Available The paper reviews the types of substrata inhabited by non-poroid resupinate Homobasidiomycetes in situ in global scale with both examples from literature sources and from observations on Belarus corticioid fungi biota. The groups of organic world colonized by corticioid basidiomata and vegetative mycelium are arboreous, semi-arboreous, and herbaceous vascular plants, Bryophyta, epiphytic coccoid algae, lichenized and non-lichenized fungi, and occasionally myxomycetes and invertebrates. The fungi occur on living, dying, and dead on all decay stages parts of organisms. Besides, the fungi are known on soil, humus, stones, artificial inorganic and synthetic materials and dung.

Mesophilic and thermophilic anaerobic digestion of primary and secondary sludge. Effect of pre-treatment at elevated temperature

DEFF Research Database (Denmark)

Gavala, Hariklia N.; Yenal, U.; Skiadas, Ioannis V.

2003-01-01

Anaerobic digestion is an appropriate technique for the treatment of sludge before final disposal and it is employed worldwide as the oldest and most important process for sludge stabilization. In general, mesophilic anaerobic digestion of sewage sludge is more widely used compared to thermophilic...... digestion. Furthermore, thermal pre-treatment is suitable for the improvement of stabilization, enhancement of dewatering of the sludge, reduction of the numbers of pathogens and could be realized at relatively low cost especially at low temperatures. The present study investigates (a) the differences...... between mesophilic and thermophilic anaerobic digestion of sludge and (b) the effect of the pretreatment at 70 degreesC on mesophilic and thermophilic anaerobic digestion of primary and secondary sludge. The pretreatment step showed very positive effect on the methane potential and production rate upon...

A constant flux of diverse thermophilic bacteria into the cold arctic seabed

DEFF Research Database (Denmark)

Hubert, Casey; Loy, Alexander; Nickel, Maren

2009-01-01

Microorganisms have been repeatedly discovered in environments that do not support their metabolic activity. Identifying and quantifying these misplaced organisms can reveal dispersal mechanisms that shape natural microbial diversity. Using endospore germination experiments, we estimated a stable...... supply of thermophilic bacteria into permanently cold Arctic marine sediment at a rate exceeding 108 spores per square meter per year. These metabolically and phylogenetically diverse Firmicutes show no detectable activity at cold in situ temperatures but rapidly mineralize organic matter by hydrolysis......, fermentation, and sulfate reduction upon induction at 50°C. The closest relatives to these bacteria come from warm subsurface petroleum reservoir and ocean crust ecosystems, suggesting that seabed fluid flow from these environments is delivering thermophiles to the cold ocean. These transport pathways may...

Genomic Encyclopedia of Fungi

Energy Technology Data Exchange (ETDEWEB)

Grigoriev, Igor

2012-08-10

Genomes of fungi relevant to energy and environment are in focus of the Fungal Genomic Program at the US Department of Energy Joint Genome Institute (JGI). Its key project, the Genomics Encyclopedia of Fungi, targets fungi related to plant health (symbionts, pathogens, and biocontrol agents) and biorefinery processes (cellulose degradation, sugar fermentation, industrial hosts), and explores fungal diversity by means of genome sequencing and analysis. Over 150 fungal genomes have been sequenced by JGI to date and released through MycoCosm (www.jgi.doe.gov/fungi), a fungal web-portal, which integrates sequence and functional data with genome analysis tools for user community. Sequence analysis supported by functional genomics leads to developing parts list for complex systems ranging from ecosystems of biofuel crops to biorefineries. Recent examples of such parts suggested by comparative genomics and functional analysis in these areas are presented here.

A comparative study of thermophilic and mesophilic anaerobic co-digestion of food waste and wheat straw: Process stability and microbial community structure shifts.

Science.gov (United States)

Shi, Xuchuan; Guo, Xianglin; Zuo, Jiane; Wang, Yajiao; Zhang, Mengyu

2018-05-01

Renewable energy recovery from organic solid waste via anaerobic digestion is a promising way to provide sustainable energy supply and eliminate environmental pollution. However, poor efficiency and operational problems hinder its wide application of anaerobic digestion. The effects of two key parameters, i.e. temperature and substrate characteristics on process stability and microbial community structure were studied using two lab-scale anaerobic reactors under thermophilic and mesophilic conditions. Both the reactors were fed with food waste (FW) and wheat straw (WS). The organic loading rates (OLRs) were maintained at a constant level of 3 kg VS/(m 3 ·d). Five different FW:WS substrate ratios were utilized in different operational phases. The synergetic effects of co-digestion improved the stability and performance of the reactors. When FW was mono-digested, both reactors were unstable. The mesophilic reactor eventually failed due to volatile fatty acid accumulation. The thermophilic reactor had better performance compared to mesophilic one. The biogas production rate of the thermophilic reactor was 4.9-14.8% higher than that of mesophilic reactor throughout the experiment. The shifts in microbial community structures throughout the experiment in both thermophilic and mesophilic reactors were investigated. With increasing FW proportions, bacteria belonging to the phylum Thermotogae became predominant in the thermophilic reactor, while the phylum Bacteroidetes was predominant in the mesophilic reactor. The genus Methanosarcina was the predominant methanogen in the thermophilic reactor, while the genus Methanothrix remained predominant in the mesophilic reactor. The methanogenesis pathway shifted from acetoclastic to hydrogenotrophic when the mesophilic reactor experienced perturbations. Moreover, the population of lignocellulose-degrading microorganisms in the thermophilic reactor was higher than those in mesophilic reactor, which explained the better

Structural and physicochemical properties of polar lipids from thermophilic archaea.

Science.gov (United States)

Ulrih, Natasa Poklar; Gmajner, Dejan; Raspor, Peter

2009-08-01

The essential general features required for lipid membranes of extremophilic archaea to fulfill biological functions are that they are in the liquid crystalline phase and have extremely low permeability of solutes that is much less temperature sensitive due to a lack of lipid-phase transition and highly branched isoprenoid chains. Many accumulated data indicate that the organism's response to extremely low pH is the opposite of that to high temperature. The high temperature adaptation does not require the tetraether lipids, while the adaptation of thermophiles to acidic environment requires the tetraether polar lipids. The presence of cyclopentane rings and the role of polar heads are not so straightforward regarding the correlations between fluidity and permeability of the lipid membrane. Due to the unique lipid structures and properties of archaeal lipids, they are a valuable resource in the development of novel biotechnological processes. This microreview focuses primarily on structural and physicochemical properties of polar lipids of (hyper)thermophilic archaea.

Bioleaching of multiple metals from contaminated sediment by moderate thermophiles.

Science.gov (United States)

Gan, Min; Jie, Shiqi; Li, Mingming; Zhu, Jianyu; Liu, Xinxing

2015-08-15

A moderately thermophilic consortium was applied in bioleaching multiple metals from contaminated sediment. The consortium got higher acidification and metals soubilization efficiency than that of the pure strains. The synergistic effect of the thermophilic consortium accelerated substrates utilization. The utilization of substrate started with sulfur in the early stage, and then the pH declined, giving rise to making use of the pyrite. Community dynamic showed that A. caldus was the predominant bacteria during the whole bioleaching process while the abundance of S. thermotolerans increased together with pyrite utilization. Solubilization efficiency of Zn, Cu, Mn and Cd reached 98%, 94%, 95%, and 89% respectively, while As, Hg, Pb was only 45%, 34%, 22%. Logistic model was used to simulate the bioleaching process, whose fitting degree was higher than 90%. Correlation analysis revealed that metal leaching was mainly an acid solubilization process. Fraction analysis revealed that metals decreased in mobility and bioavailability. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enzyme dynamics and hydrogen tunnelling in a thermophilic alcohol dehydrogenase

Science.gov (United States)

Kohen, Amnon; Cannio, Raffaele; Bartolucci, Simonetta; Klinman, Judith P.; Klinman, Judith P.

1999-06-01

Biological catalysts (enzymes) speed up reactions by many orders of magnitude using fundamental physical processes to increase chemical reactivity. Hydrogen tunnelling has increasingly been found to contribute to enzyme reactions at room temperature. Tunnelling is the phenomenon by which a particle transfers through a reaction barrier as a result of its wave-like property. In reactions involving small molecules, the relative importance of tunnelling increases as the temperature is reduced. We have now investigated whether hydrogen tunnelling occurs at elevated temperatures in a biological system that functions physiologically under such conditions. Using a thermophilic alcohol dehydrogenase (ADH), we find that hydrogen tunnelling makes a significant contribution at 65°C this is analogous to previous findings with mesophilic ADH at 25°C ( ref. 5). Contrary to predictions for tunnelling through a rigid barrier, the tunnelling with the thermophilic ADH decreases at and below room temperature. These findings provide experimental evidence for a role of thermally excited enzyme fluctuations in modulating enzyme-catalysed bond cleavage.

ANAEROBIC BIODEGRADATION OF A BIODEGRADABLE MATERIAL UNDER ANAEROBIC - THERMOPHILIC DIGESTION

Directory of Open Access Journals (Sweden)

RICARDO CAMACHO-MUÑOZ

2014-12-01

Full Text Available This paper dertermined the anaerobic biodegradation of a polymer obtained by extrusion process of native cassava starch, polylactic acid and polycaprolactone. Initially a thermophilic - methanogenic inoculum was prepared from urban solid waste. The gas final methane concentration and medium’s pH reached values of 59,6% and 7,89 respectively. The assay assembly was carried out according ASTM D5511 standard. The biodegradation percent of used materials after 15 day of digestion were: 77,49%, 61,27%, 0,31% for cellulose, sample and polyethylene respectively. Due cellulose showed biodegradation levels higher than 70% it’s deduced that the inoculum conditions were appropriate. A biodegradation level of 61,27%, 59,35% of methane concentration in sample’s evolved gas and a medium’s finale pH of 7,71 in sample’s vessels, reveal the extruded polymer´s capacity to be anaerobically degraded under thermophilic- high solid concentration conditions.

Fight Fungi with Fungi: Antifungal Properties of the Amphibian Mycobiome

Directory of Open Access Journals (Sweden)

Patrick J. Kearns

2017-12-01

Full Text Available Emerging infectious diseases caused by fungal taxa are increasing and are placing a substantial burden on economies and ecosystems worldwide. Of the emerging fungal diseases, chytridomycosis caused by the fungus Batrachochytrium dendrobatidis (hereafter Bd is linked to global amphibian declines. Amphibians have innate immunity, as well as additional resistance through cutaneous microbial communities. Despite the targeting of bacteria as potential probiotics, the role of fungi in the protection against Bd infection in unknown. We used a four-part approach, including high-throughput sequencing of bacterial and fungal communities, cultivation of fungi, Bd challenge assays, and experimental additions of probiotic to Midwife Toads (Altyes obstetricans, to examine the overlapping roles of bacterial and fungal microbiota in pathogen defense in captive bred poison arrow frogs (Dendrobates sp.. Our results revealed that cutaneous fungal taxa differed from environmental microbiota across three species and a subspecies of Dendrobates spp. frogs. Cultivation of host-associated and environmental fungi realved numerous taxa with the ability to inhibit or facilitate the growth of Bd. The abundance of cutaneous fungi contributed more to Bd defense (~45% of the fungal community, than did bacteria (~10% and frog species harbored distinct inhibitory communities that were distinct from the environment. Further, we demonstrated that a fungal probiotic therapy did not induce an endocrine-immune reaction, in contrast to bacterial probiotics that stressed amphibian hosts and suppressed antimicrobial peptide responses, limiting their long-term colonization potential. Our results suggest that probiotic strategies against amphibian fungal pathogens should, in addition to bacterial probiotics, focus on host-associated and environmental fungi such as Penicillium and members of the families Chaetomiaceae and Lasiosphaeriaceae.

Comparison of the microbial communities in solid-state anaerobic digestion (SS-AD) reactors operated at mesophilic and thermophilic temperatures.

Science.gov (United States)

Li, Yueh-Fen; Nelson, Michael C; Chen, Po-Hsu; Graf, Joerg; Li, Yebo; Yu, Zhongtang

2015-01-01

The microbiomes involved in liquid anaerobic digestion process have been investigated extensively, but the microbiomes underpinning solid-state anaerobic digestion (SS-AD) are poorly understood. In this study, microbiome composition and temporal succession in batch SS-AD reactors, operated at mesophilic or thermophilic temperatures, were investigated using Illumina sequencing of 16S rRNA gene amplicons. A greater microbial richness and evenness were found in the mesophilic than in the thermophilic SS-AD reactors. Firmicutes accounted for 60 and 82 % of the total Bacteria in the mesophilic and in the thermophilic SS-AD reactors, respectively. The genus Methanothermobacter dominated the Archaea in the thermophilic SS-AD reactors, while Methanoculleus predominated in the mesophilic SS-AD reactors. Interestingly, the data suggest syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis as an important pathway for biogas production during the thermophilic SS-AD. Canonical correspondence analysis (CCA) showed that temperature was the most influential factor in shaping the microbiomes in the SS-AD reactors. Thermotogae showed strong positive correlation with operation temperature, while Fibrobacteres, Lentisphaerae, Spirochaetes, and Tenericutes were positively correlated with daily biogas yield. This study provided new insight into the microbiome that drives SS-AD process, and the findings may help advance understanding of the microbiome in SS-AD reactors and the design and operation of SS-AD systems.

Thermophilic Dry Methane Fermentation of Distillation Residue Eluted from Ethanol Fermentation of Kitchen Waste and Dynamics of Microbial Communities.

Science.gov (United States)

Huang, Yu-Lian; Tan, Li; Wang, Ting-Ting; Sun, Zhao-Yong; Tang, Yue-Qin; Kida, Kenji

2017-01-01

Thermophilic dry methane fermentation is advantageous for feedstock with high solid content. Distillation residue with 65.1 % moisture content was eluted from ethanol fermentation of kitchen waste and subjected to thermophilic dry methane fermentation, after adjusting the moisture content to 75 %. The effect of carbon to nitrogen (C/N) ratio on thermophilic dry methane fermentation was investigated. Results showed that thermophilic dry methane fermentation could not be stably performed for >10 weeks at a C/N ratio of 12.6 and a volatile total solid (VTS) loading rate of 1 g/kg sludge/d; however, it was stably performed at a C/N ratio of 19.8 and a VTS loading rate of 3 g/kg sludge/d with 83.4 % energy recovery efficiency. Quantitative PCR analysis revealed that the number of bacteria and archaea decreased by two orders of magnitude at a C/N ratio of 12.6, whereas they were not influenced at a C/N ratio of 19.8. Microbial community analysis revealed that the relative abundance of protein-degrading bacteria increased and that of organic acid-oxidizing bacteria and acetic acid-oxidizing bacteria decreased at a C/N ratio of 12.6. Therefore, there was accumulation of NH 4 + and acetic acid, which inhibited thermophilic dry methane fermentation.

Biohydrogen production from pig slurry in a CSTR reactor system with mixed cultures under hyper-thermophilic temperature (70 oC)

International Nuclear Information System (INIS)

Kotsopoulos, Thomas A.; Fotidis, Ioannis A.; Tsolakis, Nikolaos; Martzopoulos, Gerassimos G.

2009-01-01

A continuous stirred tank reactor (CSTR) (750 cm 3 working volume) was operated with pig slurry under hyper-thermophilic (70 o C) temperature for hydrogen production. The hydraulic retention time (HRT) was 24 h and the organic loading rate was 24.9 g d -1 of volatile solid (VS). The inoculum used in the hyper-thermophilic reactor was sludge obtained from a mesophilic methanogenic reactor. The continuous feeding with active biomass (inoculum) from the mesophilic methanogenic reactor was necessary in order to achieve hydrogen production. The hyper-thermophilic reactor started to produce hydrogen after a short adapted period of 4 days. During the steady state period the mean hydrogen yield was 3.65 cm 3 g -1 of volatile solid added. The high operation temperature of the reactor enhanced the hydrolytic activity in pig slurry and increased the volatile fatty acids (VFA) production. The short HRT (24 h) and the hyper-thermophilic temperature applied in the reactor were enough to prevent methanogenesis. No pre-treatment methods or other control methods for preventing methanogenesis were necessary. Hyper-thermophilic hydrogen production was demonstrated for the first time in a CSTR system, fed with pig slurry, using mixed culture. The results indicate that this system is a promising one for biohydrogen production from pig slurry.

Marine fungi: A critique

Digital Repository Service at National Institute of Oceanography (India)

Raghukumar, S.; Raghukumar, C.

in the sea have been ignored to a large extent. However, several instances of terrestrial species of fungi, active in marine environment have been reported. The arguments to support the view that terrestrial species of fungi by virtue of their physiological...

Methanogenic H2 syntrophy among thermophiles: a model of metabolism, adaptation and survival in the subsurface

Science.gov (United States)

Topcuoglu, B. D.; Stewart, L. C.; Butterfield, D. A.; Huber, J. A.; Holden, J. F.

2016-12-01

Approximately 1 giga ton (Gt, 1015 g) of CH4 is formed globally per year from H2, CO2 and acetate through methanogenesis, largely by methanogens growing in syntrophic association with anaerobic microbes that hydrolyze and ferment biopolymers. However, our understanding of methanogenesis in hydrothermal regions of the subseafloor and potential syntrophic methanogenesis at thermophilic temperatures (i.e., >50°C) is nascent. In this study, the growth of natural assemblages of thermophilic methanogens from Axial Seamount was primarily limited by H2 availability. Heterotrophs supported thermophilic methanogenesis by H2 syntrophy in microcosm incubations of hydrothermal fluids at 55°C and 80°C supplemented with tryptone only. Based on 16S rRNA gene sequencing, only heterotrophic archaea that produce H2, H2-consuming methanogens, and sulfate reducing archaea were found in 80°C tryptone microcosms from Marker 113 vent. No bacteria were found. In 55°C tryptone microcosms, sequences were found from H2-producing bacteria and H2-consuming methanogens and sulfate-reducing bacteria. In order to model the impact of H2 syntrophy at hyperthemophilic temperatures, a co-culture was established consisting of the H2-producing hyperthermophilic heterotroph Thermococcus paralvinellae and a H2-consuming hyperthermophilic methanogen Methanocaldococcus bathoardescens. When grown alone in a chemostat, the growth rates and steady-state cell concentrations of T. paralvinellae decreased significantly when a high H2 (70 µM) background was present. H2 inhibition was ameliorated by the production of formate, but in silico modeling suggests less energetic yield for the cells. H2 syntrophy relieved H2 inhibition for both the heterotroph and the methanogenic partners. The results demonstrate that thermophilic H2 syntrophy can support methanogenesis within natural microbial assemblages and may be an important alternative energy source for thermophilic autotrophs in marine geothermal environments.

Growth media in anaerobic fermentative processes: The underestimated potential of thermophilic fermentation and anaerobic digestion.

Science.gov (United States)

Hendriks, A T W M; van Lier, J B; de Kreuk, M K

Fermentation and anaerobic digestion of organic waste and wastewater is broadly studied and applied. Despite widely available results and data for these processes, comparison of the generated results in literature is difficult. Not only due to the used variety of process conditions, but also because of the many different growth media that are used. Composition of growth media can influence biogas production (rates) and lead to process instability during anaerobic digestion. To be able to compare results of the different studies reported, and to ensure nutrient limitation is not influencing observations ascribed to process dynamics and/or reaction kinetics, a standard protocol for creating a defined growth medium for anaerobic digestion and mixed culture fermentation is proposed. This paper explains the role(s) of the different macro- and micronutrients, as well as the choices for a growth medium formulation strategy. In addition, the differences in nutrient requirements between mesophilic and thermophilic systems are discussed as well as the importance of specific trace metals regarding specific conversion routes and the possible supplementary requirement of vitamins. The paper will also give some insight into the bio-availability and toxicity of trace metals. A remarkable finding is that mesophilic and thermophilic enzymes are quite comparable at their optimum temperatures. This has consequences for the trace metal requirements of thermophiles under certain conditions. Under non-limiting conditions, the trace metal requirement of thermophilic systems is about 3 times higher than for mesophilic systems. Copyright © 2017 Elsevier Inc. All rights reserved.

Development of a gene cloning system in a fast-growing and moderately thermophilic Streptomyces species and heterologous expression of Streptomyces antibiotic biosynthetic gene clusters

Science.gov (United States)

2011-01-01

Background Streptomyces species are a major source of antibiotics. They usually grow slowly at their optimal temperature and fermentation of industrial strains in a large scale often takes a long time, consuming more energy and materials than some other bacterial industrial strains (e.g., E. coli and Bacillus). Most thermophilic Streptomyces species grow fast, but no gene cloning systems have been developed in such strains. Results We report here the isolation of 41 fast-growing (about twice the rate of S. coelicolor), moderately thermophilic (growing at both 30°C and 50°C) Streptomyces strains, detection of one linear and three circular plasmids in them, and sequencing of a 6996-bp plasmid, pTSC1, from one of them. pTSC1-derived pCWH1 could replicate in both thermophilic and mesophilic Streptomyces strains. On the other hand, several Streptomyces replicons function in thermophilic Streptomyces species. By examining ten well-sporulating strains, we found two promising cloning hosts, 2C and 4F. A gene cloning system was established by using the two strains. The actinorhodin and anthramycin biosynthetic gene clusters from mesophilic S. coelicolor A3(2) and thermophilic S. refuineus were heterologously expressed in one of the hosts. Conclusions We have developed a gene cloning and expression system in a fast-growing and moderately thermophilic Streptomyces species. Although just a few plasmids and one antibiotic biosynthetic gene cluster from mesophilic Streptomyces were successfully expressed in thermophilic Streptomyces species, we expect that by utilizing thermophilic Streptomyces-specific promoters, more genes and especially antibiotic genes clusters of mesophilic Streptomyces should be heterologously expressed. PMID:22032628

Synergistic and Dose-Controlled Regulation of Cellulase Gene Expression in Penicillium oxalicum.

Science.gov (United States)

Li, Zhonghai; Yao, Guangshan; Wu, Ruimei; Gao, Liwei; Kan, Qinbiao; Liu, Meng; Yang, Piao; Liu, Guodong; Qin, Yuqi; Song, Xin; Zhong, Yaohua; Fang, Xu; Qu, Yinbo

2015-09-01

Filamentous fungus Penicillium oxalicum produces diverse lignocellulolytic enzymes, which are regulated by the combinations of many transcription factors. Here, a single-gene disruptant library for 470 transcription factors was constructed and systematically screened for cellulase production. Twenty transcription factors (including ClrB, CreA, XlnR, Ace1, AmyR, and 15 unknown proteins) were identified to play putative roles in the activation or repression of cellulase synthesis. Most of these regulators have not been characterized in any fungi before. We identified the ClrB, CreA, XlnR, and AmyR transcription factors as critical dose-dependent regulators of cellulase expression, the core regulons of which were identified by analyzing several transcriptomes and/or secretomes. Synergistic and additive modes of combinatorial control of each cellulase gene by these regulatory factors were achieved, and cellulase expression was fine-tuned in a proper and controlled manner. With one of these targets, the expression of the major intracellular β-glucosidase Bgl2 was found to be dependent on ClrB. The Bgl2-deficient background resulted in a substantial gene activation by ClrB and proved to be closely correlated with the relief of repression mediated by CreA and AmyR during cellulase induction. Our results also signify that probing the synergistic and dose-controlled regulation mechanisms of cellulolytic regulators and using it for reconstruction of expression regulation network (RERN) may be a promising strategy for cellulolytic fungi to develop enzyme hyper-producers. Based on our data, ClrB was identified as focal point for the synergistic activation regulation of cellulase expression by integrating cellulolytic regulators and their target genes, which refined our understanding of transcriptional-regulatory network as a "seesaw model" in which the coordinated regulation of cellulolytic genes is established by counteracting activators and repressors.

Autophagy in plant pathogenic fungi.

Science.gov (United States)

Liu, Xiao-Hong; Xu, Fei; Snyder, John Hugh; Shi, Huan-Bin; Lu, Jian-Ping; Lin, Fu-Cheng

2016-09-01

Autophagy is a conserved cellular process that degrades cytoplasmic constituents in vacuoles. Plant pathogenic fungi develop special infection structures and/or secrete a range of enzymes to invade their plant hosts. It has been demonstrated that monitoring autophagy processes can be extremely useful in visualizing the sequence of events leading to pathogenicity of plant pathogenic fungi. In this review, we introduce the molecular mechanisms involved in autophagy. In addition, we explore the relationship between autophagy and pathogenicity in plant pathogenic fungi. Finally, we discuss the various experimental strategies available for use in the study of autophagy in plant pathogenic fungi. Copyright © 2016 Elsevier Ltd. All rights reserved.

Widespread Disulfide Bonding in Proteins from Thermophilic Archaea

OpenAIRE

Jorda, Julien; Yeates, Todd O.

2011-01-01

Disulfide bonds are generally not used to stabilize proteins in the cytosolic compartments of bacteria or eukaryotic cells, owing to the chemically reducing nature of those environments. In contrast, certain thermophilic archaea use disulfide bonding as a major mechanism for protein stabilization. Here, we provide a current survey of completely sequenced genomes, applying computational methods to estimate the use of disulfide bonding across the Archaea. Microbes belonging to the Crenarchaea...

Domestic sewage sludge composting in a rotary drum reactor: optimizing the thermophilic stage.

Science.gov (United States)

Rodríguez, Luis; Cerrillo, María I; García-Albiach, Valentín; Villaseñor, José

2012-12-15

The aim of this paper was to study the influence of four process variables (turning frequency, gas-phase oxygen level, type of bulking agent and sludge/bulking agent mixing ratio) on the performance of the sewage sludge composting process using a rotary drum pilot scale reactor, in order to optimize the thermophilic stage and reduce the processing time. Powdered sawdust, wood shavings, wood chips, prunings waste and straw were used as bulking agents and the thermophilic stage temperature profile was used as the main indicator for gauging if the composting process was developing correctly. Our results showed that a 12 h(-1) turning frequency and an oxygen concentration of 10% were the optimal conditions for the composting process to develop. The best results were obtained by mixing the sewage sludge with wood shavings in a 3:1 w/w ratio (on a wet basis), which adapted the initial moisture content and porosity to an optimal range and led to a maximum temperature of 70 °C being reached thus ensuring the complete removal of pathogens. Moisture, C:N ratio, pH, organic matter, heavy metals, pathogens and stability were all analysed for every mixture obtained at the end of the thermophilic stage. These parameters were compared with the limits established by the Spanish regulation on fertilizers (RD 824/2005) in order to assess if the compost obtained could be used on agricultural soils. The right combination of having optimal process variables combined with an appropriate reactor design allowed the thermophilic stage of the composting process to be speeded up, hence obtaining a compost product, after just two weeks of processing that (with the exception of the moisture content) complied with the Spanish legal requirements for fertilizers, without requiring a later maturation stage. Copyright © 2012 Elsevier Ltd. All rights reserved.

Design of A solar Thermophilic Anaerobic Reactor for Small Farms

NARCIS (Netherlands)

Mashad, El H.; Loon, van W.K.P.; Zeeman, G.; Bot, G.P.A.; Lettinga, G.

2004-01-01

A 10 m(3) completely stirred tank reactor has been designed for anaerobic treatment of liquid cow manure under thermophilic conditions (50degreesC), using a solar heating system mounted on the reactor roof. Simulation models for two systems have been developed. The first system consists of loose

Proteomics of Filamentous Fungi

NARCIS (Netherlands)

Passel, van M.W.J.; Schaap, P.J.; Graaff, de L.H.

2013-01-01

Filamentous fungi, such as Aspergillus niger and Aspergillus oryzae traditionally have had an important role in providing enzymes and enzyme cocktails that are used in food industry. In recent years the genome sequences of many filamentous fungi have become available. This combined with

Hydrolytic activities of extracellular enzymes in thermophilic and mesophilic anaerobic sequencing-batch reactors treating organic fractions of municipal solid wastes.

Science.gov (United States)

Kim, Hyun-Woo; Nam, Joo-Youn; Kang, Seok-Tae; Kim, Dong-Hoon; Jung, Kyung-Won; Shin, Hang-Sik

2012-04-01

Extracellular enzymes offer active catalysis for hydrolysis of organic solid wastes in anaerobic digestion. To evidence the quantitative significance of hydrolytic enzyme activities for major waste components, track studies of thermophilic and mesophilic anaerobic sequencing-batch reactors (TASBR and MASBR) were conducted using a co-substrate of real organic wastes. During 1day batch cycle, TASBR showed higher amylase activity for carbohydrate (46%), protease activity for proteins (270%), and lipase activity for lipids (19%) than MASBR. In particular, the track study of protease identified that thermophilic anaerobes degraded protein polymers much more rapidly. Results revealed that differences in enzyme activities eventually affected acidogenic and methanogenic performances. It was demonstrated that the superior nature of enzymatic capability at thermophilic condition led to successive high-rate acidogenesis and 32% higher CH(4) recovery. Consequently, these results evidence that the coupling thermophilic digestion with sequencing-batch operation is a viable option to promote enzymatic hydrolysis of organic particulates. Copyright © 2012 Elsevier Ltd. All rights reserved.

Assembly and multiple gene expression of thermophilic enzymes in Escherichia coli for in vitro metabolic engineering.

Science.gov (United States)

Ninh, Pham Huynh; Honda, Kohsuke; Sakai, Takaaki; Okano, Kenji; Ohtake, Hisao

2015-01-01

In vitro reconstitution of an artificial metabolic pathway is an emerging approach for the biocatalytic production of industrial chemicals. However, several enzymes have to be separately prepared (and purified) for the construction of an in vitro metabolic pathway, thereby limiting the practical applicability of this approach. In this study, genes encoding the nine thermophilic enzymes involved in a non-ATP-forming chimeric glycolytic pathway were assembled in an artificial operon and co-expressed in a single recombinant Escherichia coli strain. Gene expression levels of the thermophilic enzymes were controlled by their sequential order in the artificial operon. The specific activities of the recombinant enzymes in the cell-free extract of the multiple-gene-expression E. coli were 5.0-1,370 times higher than those in an enzyme cocktail prepared from a mixture of single-gene-expression strains, in each of which a single one of the nine thermophilic enzymes was overproduced. Heat treatment of a crude extract of the multiple-gene-expression cells led to the denaturation of indigenous proteins and one-step preparation of an in vitro synthetic pathway comprising only a limited number of thermotolerant enzymes. Coupling this in vitro pathway with other thermophilic enzymes including the H2 O-forming NADH oxidase or the malate/lactate dehydrogenase facilitated one-pot conversion of glucose to pyruvate or lactate, respectively. © 2014 Wiley Periodicals, Inc.

Thermophilic Bacteria Colony Growwth and its Consequences in the Food Industry

Czech Academy of Sciences Publication Activity Database

Melzoch, K.; Votruba, Jaroslav; Sekavová, B.; Piterková, L.; Rychtera, M.

2004-01-01

Roč. 22, č. 1 (2004), s. 1-8 ISSN 1212-1800 R&D Projects: GA ČR GA525/03/0375 Institutional research plan: CEZ:AV0Z5020903 Keywords : thermophilic bacteria * colony growth Subject RIV: EE - Microbiology, Virology

Filamentous Fungi.

Science.gov (United States)

Powers-Fletcher, Margaret V; Kendall, Brian A; Griffin, Allen T; Hanson, Kimberly E

2016-06-01

Filamentous mycoses are often associated with significant morbidity and mortality. Prompt diagnosis and aggressive treatment are essential for good clinical outcomes in immunocompromised patients. The host immune response plays an essential role in determining the course of exposure to potential fungal pathogens. Depending on the effectiveness of immune response and the burden of organism exposure, fungi can either be cleared or infection can occur and progress to a potentially fatal invasive disease. Nonspecific cellular immunity (i.e., neutrophils, natural killer [NK] cells, and macrophages) combined with T-cell responses are the main immunologic mechanisms of protection. The most common potential mold pathogens include certain hyaline hyphomycetes, endemic fungi, the Mucorales, and some dematiaceous fungi. Laboratory diagnostics aimed at detecting and differentiating these organisms are crucial to helping clinicians make informed decisions about treatment. The purpose of this chapter is to provide an overview of the medically important fungal pathogens, as well as to discuss the patient characteristics, antifungal-therapy considerations, and laboratory tests used in current clinical practice for the immunocompromised host.

Seasonal Variability of Thermophilic Campylobacter Spp. in Raw Milk Sold by Automatic Vending Machines in Lombardy Region.

Science.gov (United States)

Bertasi, Barbara; Losio, Marina Nadia; Daminelli, Paolo; Finazzi, Guido; Serraino, Andrea; Piva, Silvia; Giacometti, Federica; Massella, Elisa; Ostanello, Fabio

2016-06-03

In temperate climates, a seasonal trend was observed in the incidence of human campylobacteriosis cases, with peaks reported in spring and autumn in some countries, or in summer in others; a similar trend was observed in Campylobacter spp. dairy cattle faecal shedding, suggesting that cattle may play a role in the seasonal peak of human infection. The objectives of this study were to assess if a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk exists and to evaluate a possible relation between this and the increase of human campylobacteriosis incidence in summer months. The results showed a mean prevalence of 1.6% of milk samples positive for thermophilic Campylobacter spp. with a wide range (0.0-3.1%) in different months during the three years considered. The statistical analysis showed a significant difference (P<0.01) of the prevalence of positive samples for thermophilic Campylobacter spp. between warmer and cooler months (2.3 vs 0.6%). The evidence of a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk sold for direct consumption, with an increase of the prevalence in warmer months, may represent one of the possible links between seasonal trend in cattle faecal shedding and seasonal trend in human campylobacteriosis.

Lectins in human pathogenic fungi.

Science.gov (United States)

Gallegos, Belém; Martínez, Ruth; Pérez, Laura; Del Socorro Pina, María; Perez, Eduardo; Hernández, Pedro

2014-01-01

Lectins are carbohydrate-binding proteins widely distributed in nature. They constitute a highly diverse group of proteins consisting of many different protein families that are, in general, structurally unrelated. In the last few years, mushroom and other fungal lectins have attracted wide attention due to their antitumour, antiproliferative and immunomodulatory activities. The present mini-review provides concise information about recent developments in understanding lectins from human pathogenic fungi. A bibliographic search was performed in the Science Direct and PubMed databases, using the following keywords "lectin", "fungi", "human" and "pathogenic". Lectins present in fungi have been classified; however, the role played by lectins derived from human pathogenic fungi in infectious processes remains uncertain; thus, this is a scientific field requiring more research. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012). Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

Performance of thermophilic anaerobic digesters using inoculum mixes with enhanced methanogenic diversity

KAUST Repository

Ghanimeh, Sophia; El-Fadel, Mutasem; Saikaly, Pascal

2017-01-01

Reportedly, various mixes of seeds were quasi-randomly selected to startup anaerobic digesters. In contrast, this study examines the impact of inoculating thermophilic anaerobic digesters with a designed mix of non-acclimated seeds based

Global transport of thermophilic bacteria in atmospheric dust.

Science.gov (United States)

Perfumo, Amedea; Marchant, Roger

2010-04-01

Aerosols from dust storms generated in the Sahara-Sahel desert area of Africa are transported north over Europe and periodically result in dry dust precipitation in the Mediterranean region. Samples of dust collected in Turkey and Greece following two distinct desert storm events contained viable thermophilic organisms of the genus Geobacillus, namely G. thermoglucosidasius and G. thermodenitrificans, and the recently reclassified Aeribacillus pallidus (formerly Geobacillus pallidus). We present here evidence that African dust storms create an atmospheric bridge between distant geographical regions and that they are also probably the source of thermophilic geobacilli later deposited over northern Europe by rainfall or dust plumes themselves. The same organisms (99% similarity in the 16S rDNA sequence) were found in dust collected in the Mediterranean region and inhabiting cool soils in Northern Ireland. This study also contributes new insights to the taxonomic identification of Geobacillus sp. Attempts to identify these organisms using 16S rRNA gene sequences have revealed that they contain multiple and diverse copies of the ribosomal RNA operon (up to 10 copies with nine different sequences), which dictates care in interpreting data about the systematics of this genus. © 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.

Production of Sporotrichum thermophile xylanase by solid state fermentation utilizing deoiled Jatropha curcas seed cake and its application in xylooligosachharide synthesis.

Science.gov (United States)

Sadaf, Ayesha; Khare, S K

2014-02-01

De-oiled Jatropha curcas seed cake, a plentiful by-product of biodiesel industry was used as substrate for the production of a useful xylanase from Sporotrichum thermophile in solid state fermentation. Under the optimized conditions, 1025U xylanase/g (deoiled seed cake) was produced. The xylanase exhibited half life of 4h at 45°C and 71.44min at 50°C respectively. It was stable in a broad pH range of 7.0-11.0. Km and Vmax were 12.54mg/ml and 454.5U/ml/min respectively. S. thermophile xylanase is an endoxylanase free of exoxylanase activity, hence advantageous for xylan hydrolysis to produce xylooligosachharides. Hydrolysis of oat spelt xylan by S. thermophile xylanase yielded 73% xylotetraose, 15.4% xylotriose and 10% xylobiose. The S. thermophile endoxylanase thus seem potentially useful in the food industries. Copyright © 2013 Elsevier Ltd. All rights reserved.

Thermostable 𝜶-Amylase Activity from Thermophilic Bacteria Isolated from Bora Hot Spring, Central Sulawesi

Science.gov (United States)

Gazali, F. M.; Suwastika, I. N.

2018-03-01

α-Amylase is one of the most important enzyme in biotechnology field, especially in industrial application. Thermostability of α-Amylase produced by thermophilic bacteria improves industrial process of starch degradation in starch industry. The present study were concerned to the characterization of α-Amylase activity from indigenous thermophilic bacteria isolated from Bora hot spring, Central Sulawesi. There were 18 isolates which had successfully isolated from 90°C sediment samples of Bora hot spring and 13 of them showed amylolytic activity. The α-Amylase activity was measured qualitatively at starch agar and quantitatively based on DNS (3,5-Dinitrosalicylic acid) methods, using maltose as standard solution. Two isolates (out of 13 amylolytic bacteria), BR 002 and BR 015 showed amylolytic index of 0.8 mm and 0.5 mm respectively, after being incubated at 55°C in the 0.002% Starch Agar Medium. The α-Amylase activity was further characterized quantitatively which includes the optimum condition of pH and temperature of α-Amylase crude enzyme from each isolate. To our knowledge, this is the first report on isolation and characterization of a thermostable α-Amylase from thermophilic bacteria isolated from Central Sulawesi particularly from Bora hot spring.

Isolation of aerobic cultivable cellulolytic bacteria from different regions of the gastrointestinal tract of giant land snail Achatina fulica

Directory of Open Access Journals (Sweden)

Guilherme L. Pinheiro

2015-08-01

Full Text Available The enzymatic hydrolysis of cellulose by cellulases is one of the major limiting steps in the conversion of lignocellulosic biomass to yield bioethanol. To overcome this hindrance, significant efforts are underway to identify novel cellulases. The snail Achatina fulica is a gastropod with high cellulolytic activity, mainly due to the abundance of glycoside hydrolases produced by both the animal and its resident microbiota. In this study, we partially assessed the cellulolytic bacterial diversity inside the gastrointestinal tract of A. fulica by culture-dependent methods and evaluated the hydrolytic repertoire of the isolates. Forty bacterial isolates were recovered from distinct segments of the snail gut and identified to the genus level by 16S rRNA gene sequence analysis. Additional phenotypic characterization was performed using biochemical tests provided by the Vitek2 identification system. The overall enzymatic repertoire of the isolated strains was investigated by enzymatic plate assays, containing the following substrates: powdered sugarcane bagasse, carboxymethylcellulose (CMC, p-nitrophenyl-b-D-glucopyranoside (pNPG, p-nitrophenyl-b-D-cellobioside (pNPC, 4-methylumbelliferyl-b-D-glucopyranoside (MUG, 4-methylumbelliferyl-b-D-cellobioside (MUC and 4-methylumbelliferyl-b-D-xylopyranoside (MUX. Our results indicate that the snail Achatina fulica is an attractive source of cultivable bacteria that showed to be valuable resources for the production of different types of biomass-degrading enzymes.

Isolation of aerobic cultivable cellulolytic bacteria from different regions of the gastrointestinal tract of giant land snail Achatina fulica.

Science.gov (United States)

Pinheiro, Guilherme L; Correa, Raquel F; Cunha, Raquel S; Cardoso, Alexander M; Chaia, Catia; Clementino, Maysa M; Garcia, Eloi S; de Souza, Wanderley; Frasés, Susana

2015-01-01

The enzymatic hydrolysis of cellulose by cellulases is one of the major limiting steps in the conversion of lignocellulosic biomass to yield bioethanol. To overcome this hindrance, significant efforts are underway to identify novel cellulases. The snail Achatina fulica is a gastropod with high cellulolytic activity, mainly due to the abundance of glycoside hydrolases produced by both the animal and its resident microbiota. In this study, we partially assessed the cellulolytic aerobic bacterial diversity inside the gastrointestinal tract of A. fulica by culture-dependent methods and evaluated the hydrolytic repertoire of the isolates. Forty bacterial isolates were recovered from distinct segments of the snail gut and identified to the genus level by 16S rRNA gene sequence analysis. Additional phenotypic characterization was performed using biochemical tests provided by the Vitek2 identification system. The overall enzymatic repertoire of the isolated strains was investigated by enzymatic plate assays, containing the following substrates: powdered sugarcane bagasse, carboxymethylcellulose (CMC), p-nitrophenyl-β-D-glucopyranoside (pNPG), p-nitrophenyl-β-D-cellobioside (pNPC), 4-methylumbelliferyl-β-D-glucopyranoside (MUG), 4-methylumbelliferyl-β-D-cellobioside (MUC), and 4-methylumbelliferyl-β-D-xylopyranoside (MUX). Our results indicate that the snail A. fulica is an attractive source of cultivable bacteria that showed to be valuable resources for the production of different types of biomass-degrading enzymes.

[Indiscriminate use of Latin name for natural Cordyceps sinensis insect-fungi complex and multiple Ophiocordyceps sinensis fungi].

Science.gov (United States)

Yao, Yi-Sang; Zhu, Jia-Shi

2016-04-01

Natural Cordyceps sinensis(Dongchongxiacao) is an insect-fungi complex containing multiple Ophiocordyceps sinensis(≡Cordyceps sinensis) fungi and dead body of larva of the family of Hepialidae. But natural C. sinensis and O. sinensis fungi use the same Latin name, resulting in uncertainty of the specific meaning, even disturbing the formulation and implementation of governmental policies and regulations, and influencing consumer psychology onthe market. This paper reviews the history and current status of the indiscriminate use of the Latin name O. sinensis for both the natural insect-fungi complex C. sinensis and O. sinensis fungi and lists the rename suggetions. Some scholars suggested using the term O. sinensis for the fungi and renaming the natural C. sinensis "Chinese cordyceps". Others suggested renaming the natural C. sinensis "Ophiocordyceps & Hepialidae". Both suggestions have not reached general consensus due to various academic concerns. This paper also reviews the exacerbation of the academic uncertainties when forcing implementing the 2011 Amsterdam Declaration "One Fungus=One Name" under the academic debate. Joint efforts of mycological, zoological and botany-TCM taxonomists and properly initiating the dispute systems offered by International Mycology Association may solve the debate on the indiscriminate use of the Latin name O.sinensis for the natural insect-fungi complex,the teleomorph and anamorph(s) of O. sinensis fungi. Copyright© by the Chinese Pharmaceutical Association.

Establishment of thermophilic anaerobic terephthalic acid degradation system through one-step temperature increase startup strategy - Revealed by Illumina Miseq Sequencing.

Science.gov (United States)

Ma, Kai-Li; Li, Xiang-Kun; Wang, Ke; Meng, Ling-Wei; Liu, Gai-Ge; Zhang, Jie

2017-10-01

Over recent years, thermophilic digestion was constantly focused owing to its various advantage over mesophilic digestion. Notably, the startup approach of thermophilic digester needs to be seriously considered as unsuitable startup ways may result in system inefficiency. In this study, one-step temperature increase startup strategy from 37 °C to 55 °C was applied to establish a thermophilic anaerobic system treating terephthalic acid (TA) contained wastewater, meanwhile, the archaeal and bacterial community compositions at steady periods of 37 °C and 55 °C during the experimental process was also compared using Illumina Miseq Sequencing. The process operation demonstrated that the thermophilic TA degradation system was successfully established at 55 °C with over 95% COD reduction. For archaea community, the elevation of operational temperature from 37 °C to 55 °C accordingly increase the enrichment of hydrogenotrophic methanogens but decrease the abundance of the acetotrophic ones. While for bacterial community, the taxonomic analysis suggested that Syntrophorhabdus (27.40%) was the dominant genus promoting the efficient TA degradation under mesophilic condition, whereas OPB95 (24.99%) and TA06 (14.01%) related populations were largely observed and probably take some crucial role in TA degradation under thermophilic condition. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of bioaugmentation by cellulolytic bacteria enriched from sheep rumen on methane production from wheat straw.

Science.gov (United States)

Ozbayram, E Gozde; Kleinsteuber, Sabine; Nikolausz, Marcell; Ince, Bahar; Ince, Orhan

2017-08-01

The aim of this study was to determine the potential of bioaugmentation with cellulolytic rumen microbiota to enhance the anaerobic digestion of lignocellulosic feedstock. An anaerobic cellulolytic culture was enriched from sheep rumen fluid using wheat straw as substrate under mesophilic conditions. To investigate the effects of bioaugmentation on methane production from straw, the enrichment culture was added to batch reactors in proportions of 2% (Set-1) and 4% (Set-2) of the microbial cell number of the standard inoculum slurry. The methane production in the bioaugmented reactors was higher than in the control reactors. After 30 days of batch incubation, the average methane yield was 154 mL N CH 4 g VS -1 in the control reactors. Addition of 2% enrichment culture did not enhance methane production, whereas in Set-2 the methane yield was increased by 27%. The bacterial communities were examined by 454 amplicon sequencing of 16S rRNA genes, while terminal restriction fragment length polymorphism (T-RFLP) fingerprinting of mcrA genes was applied to analyze the methanogenic communities. The results highlighted that relative abundances of Ruminococcaceae and Lachnospiraceae increased during the enrichment. However, Cloacamonaceae, which were abundant in the standard inoculum, dominated the bacterial communities of all batch reactors. T-RFLP profiles revealed that Methanobacteriales were predominant in the rumen fluid, whereas the enrichment culture was dominated by Methanosarcinales. In the batch rectors, the most abundant methanogens were affiliated to Methanobacteriales and Methanomicrobiales. Our results suggest that bioaugmentation with sheep rumen enrichment cultures can enhance the performance of digesters treating lignocellulosic feedstock. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of thermo-tolerant actinomycetes inoculation on cellulose degradation and the formation of humic substances during composting.

Science.gov (United States)

Zhao, Yi; Zhao, Yue; Zhang, Zhechao; Wei, Yuquan; Wang, Huan; Lu, Qian; Li, Yanjie; Wei, Zimin

2017-10-01

The inoculum containing four cellulolytic thermophilic actinomycetes was screened from compost samples, and was inoculated into co-composting during different inoculation phases. The effect of different inoculation phases on cellulose degradation, humic substances formation and the relationship between inoculation and physical-chemical parameters was determined. The results revealed that inoculation at different phases of composting improved cellulase activities, accelerated the degradation of cellulose, increased the content of humic substances and influenced the structure of actinomycetic community, but there were significant differences between different inoculation phases. Redundancy analysis showed that the different inoculation phases had different impacts on the relationship between exogenous actinobacteria and physical-chemical parameters. Therefore, based on the promoting effort of inoculation in thermophilic phase of composting for the formation of humic substances, we suggested an optimized inoculation strategy to increase the content of humic substances, alleviate CO 2 emission during composting. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comparing mesophilic and thermophilic anaerobic digestion of chicken manure: Microbial community dynamics and process resilience

International Nuclear Information System (INIS)

Niu, Qigui; Takemura, Yasuyuki; Kubota, Kengo; Li, Yu-You

2015-01-01

Highlights: • Microbial community dynamics and process functional resilience were investigated. • The threshold of TAN in mesophilic reactor was higher than the thermophilic reactor. • The recoverable archaeal community dynamic sustained the process resilience. • Methanosarcina was more sensitive than Methanoculleus on ammonia inhibition. • TAN and FA effects the dynamic of hydrolytic and acidogenic bacteria obviously. - Abstract: While methane fermentation is considered as the most successful bioenergy treatment for chicken manure, the relationship between operational performance and the dynamic transition of archaeal and bacterial communities remains poorly understood. Two continuous stirred-tank reactors were investigated under thermophilic and mesophilic conditions feeding with 10%TS. The tolerance of thermophilic reactor on total ammonia nitrogen (TAN) was found to be 8000 mg/L with free ammonia (FA) 2000 mg/L compared to 16,000 mg/L (FA1500 mg/L) of mesophilic reactor. Biomethane production was 0.29 L/gV S in in the steady stage and decreased following TAN increase. After serious inhibition, the mesophilic reactor was recovered successfully by dilution and washing stratagem compared to the unrecoverable of thermophilic reactor. The relationship between the microbial community structure, the bioreactor performance and inhibitors such as TAN, FA, and volatile fatty acid was evaluated by canonical correspondence analysis. The performance of methanogenic activity and substrate removal efficiency were changed significantly correlating with the community evenness and phylogenetic structure. The resilient archaeal community was found even after serious inhibition in both reactors. Obvious dynamics of bacterial communities were observed in acidogenic and hydrolytic functional bacteria following TAN variation in the different stages

Comparing mesophilic and thermophilic anaerobic digestion of chicken manure: Microbial community dynamics and process resilience

Energy Technology Data Exchange (ETDEWEB)

Niu, Qigui; Takemura, Yasuyuki; Kubota, Kengo [Department of Civil and Environmental Engineering, Graduate School of Engineering Tohoku University, 6-6-06 Aza-Aoba, Aramaki, Aoba-ku, Sendai, Miyagi 980-8579 (Japan); Li, Yu-You, E-mail: yyli@epl1.civil.tohoku.ac.jp [Department of Civil and Environmental Engineering, Graduate School of Engineering Tohoku University, 6-6-06 Aza-Aoba, Aramaki, Aoba-ku, Sendai, Miyagi 980-8579 (Japan); Key Lab of Northwest Water Resource, Environment and Ecology, MOE, Xi’an University of Architecture and Technology, Xi’an (China)

2015-09-15

Highlights: • Microbial community dynamics and process functional resilience were investigated. • The threshold of TAN in mesophilic reactor was higher than the thermophilic reactor. • The recoverable archaeal community dynamic sustained the process resilience. • Methanosarcina was more sensitive than Methanoculleus on ammonia inhibition. • TAN and FA effects the dynamic of hydrolytic and acidogenic bacteria obviously. - Abstract: While methane fermentation is considered as the most successful bioenergy treatment for chicken manure, the relationship between operational performance and the dynamic transition of archaeal and bacterial communities remains poorly understood. Two continuous stirred-tank reactors were investigated under thermophilic and mesophilic conditions feeding with 10%TS. The tolerance of thermophilic reactor on total ammonia nitrogen (TAN) was found to be 8000 mg/L with free ammonia (FA) 2000 mg/L compared to 16,000 mg/L (FA1500 mg/L) of mesophilic reactor. Biomethane production was 0.29 L/gV S{sub in} in the steady stage and decreased following TAN increase. After serious inhibition, the mesophilic reactor was recovered successfully by dilution and washing stratagem compared to the unrecoverable of thermophilic reactor. The relationship between the microbial community structure, the bioreactor performance and inhibitors such as TAN, FA, and volatile fatty acid was evaluated by canonical correspondence analysis. The performance of methanogenic activity and substrate removal efficiency were changed significantly correlating with the community evenness and phylogenetic structure. The resilient archaeal community was found even after serious inhibition in both reactors. Obvious dynamics of bacterial communities were observed in acidogenic and hydrolytic functional bacteria following TAN variation in the different stages.

Molecular interactions within the halophilic, thermophilic, and mesophilic prokaryotic ribosomal complexes: clues to environmental adaptation.

Science.gov (United States)

Mallik, Saurav; Kundu, Sudip

2015-01-01

Using the available crystal structures of 50S ribosomal subunits from three prokaryotic species: Escherichia coli (mesophilic), Thermus thermophilus (thermophilic), and Haloarcula marismortui (halophilic), we have analyzed different structural features of ribosomal RNAs (rRNAs), proteins, and of their interfaces. We have correlated these structural features with the environmental adaptation strategies of the corresponding species. While dense intra-rRNA packing is observed in thermophilic, loose intra-rRNA packing is observed in halophilic (both compared to mesophilic). Interestingly, protein-rRNA interfaces of both the extremophiles are densely packed compared to that of the mesophilic. The intersubunit bridge regions are almost devoid of cavities, probably ensuring the proper formation of each bridge (by not allowing any loosely packed region nearby). During rRNA binding, the ribosomal proteins experience some structural transitions. Here, we have analyzed the intrinsically disordered and ordered regions of the ribosomal proteins, which are subjected to such transitions. The intrinsically disordered and disorder-to-order transition sites of the thermophilic and mesophilic ribosomal proteins are simultaneously (i) highly conserved and (ii) slowly evolving compared to rest of the protein structure. Although high conservation is observed at such sites of halophilic ribosomal proteins, but slow rate of evolution is absent. Such differences between thermophilic, mesophilic, and halophilic can be explained from their environmental adaptation strategy. Interestingly, a universal biophysical principle evident by a linear relationship between the free energy of interface formation, interface area, and structural changes of r-proteins during assembly is always maintained, irrespective of the environmental conditions.

Comparative genotyping of Clostridium thermocellum strains isolated from biogas plants: genetic markers and characterization of cellulolytic potential.

Science.gov (United States)

Koeck, Daniela E; Zverlov, Vladimir V; Liebl, Wolfgang; Schwarz, Wolfgang H

2014-07-01

Clostridium thermocellum is among the most prevalent of known anaerobic cellulolytic bacteria. In this study, genetic and phenotypic variations among C. thermocellum strains isolated from different biogas plants were determined and different genotyping methods were evaluated on these isolates. At least two C. thermocellum strains were isolated independently from each of nine different biogas plants via enrichment on cellulose. Various DNA-based genotyping methods such as ribotyping, RAPD (Random Amplified Polymorphic DNA) and VNTR (Variable Number of Tandem Repeats) were applied to these isolates. One novel approach - the amplification of unknown target sequences between copies of a previously discovered Random Inserted Mobile Element (RIME) - was also tested. The genotyping method with the highest discriminatory power was found to be the amplification of the sequences between the insertion elements, where isolates from each biogas plant yielded a different band pattern. Cellulolytic potentials, optimal growth conditions and substrate spectra of all isolates were characterized to help identify phenotypic variations. Irrespective of the genotyping method used, the isolates from each individual biogas plant always exhibited identical patterns. This is suggestive of a single C. thermocellum strain exhibiting dominance in each biogas plant. The genotypic groups reflect the results of the physiological characterization of the isolates like substrate diversity and cellulase activity. Conversely, strains isolated across a range of biogas plants differed in their genotyping results and physiological properties. Both strains isolated from one biogas plant had the best specific cellulose-degrading properties and might therefore achieve superior substrate utilization yields in biogas fermenters. Copyright © 2014 Elsevier GmbH. All rights reserved.

Thermophilic Sulfate Reduction in Hydrothermal Sediment of Lake Tanganyika, East-Africa

DEFF Research Database (Denmark)

ELSGAARD, L.; PRIEUR, D.; MUKWAYA, GM

1994-01-01

at up to 70 and 75 degrees C, with optima at 63 and 71 degrees C, respectively. Several sporulating thermophilic enrichments were morphologically similar to Desulfotomaculum spp. Dissimilatory sulfate reduction in the studied hydrothermal area of Lake Tanganyika apparently has an upper temperature limit...

Thermophilic archaea activate butane via alkyl-coenzyme M formation.

Science.gov (United States)

Laso-Pérez, Rafael; Wegener, Gunter; Knittel, Katrin; Widdel, Friedrich; Harding, Katie J; Krukenberg, Viola; Meier, Dimitri V; Richter, Michael; Tegetmeyer, Halina E; Riedel, Dietmar; Richnow, Hans-Hermann; Adrian, Lorenz; Reemtsma, Thorsten; Lechtenfeld, Oliver J; Musat, Florin

2016-11-17

The anaerobic formation and oxidation of methane involve unique enzymatic mechanisms and cofactors, all of which are believed to be specific for C 1 -compounds. Here we show that an anaerobic thermophilic enrichment culture composed of dense consortia of archaea and bacteria apparently uses partly similar pathways to oxidize the C 4 hydrocarbon butane. The archaea, proposed genus 'Candidatus Syntrophoarchaeum', show the characteristic autofluorescence of methanogens, and contain highly expressed genes encoding enzymes similar to methyl-coenzyme M reductase. We detect butyl-coenzyme M, indicating archaeal butane activation analogous to the first step in anaerobic methane oxidation. In addition, Ca. Syntrophoarchaeum expresses the genes encoding β-oxidation enzymes, carbon monoxide dehydrogenase and reversible C 1 methanogenesis enzymes. This allows for the complete oxidation of butane. Reducing equivalents are seemingly channelled to HotSeep-1, a thermophilic sulfate-reducing partner bacterium known from the anaerobic oxidation of methane. Genes encoding 16S rRNA and methyl-coenzyme M reductase similar to those identifying Ca. Syntrophoarchaeum were repeatedly retrieved from marine subsurface sediments, suggesting that the presented activation mechanism is naturally widespread in the anaerobic oxidation of short-chain hydrocarbons.

Isolation and Screening of Thermophilic Bacilli from Compost for Electrotransformation and Fermentation: Characterization of Bacillus smithii ET 138 as a New Biocatalyst

NARCIS (Netherlands)

Bosma, E.F.; Weijer, van de A.H.P.; Daas, M.J.A.; Oost, van der J.; Vos, de W.M.; Kranenburg, van R.

2015-01-01

Thermophilic bacteria are regarded as attractive production organisms for cost-efficient conversion of renewable resources to green chemicals, but their genetic accessibility is a major bottleneck in developing them into versatile platform organisms. In this study, we aimed to isolate thermophilic,

Fungi as a Source of Food.

Science.gov (United States)

Dupont, Joëlle; Dequin, Sylvie; Giraud, Tatiana; Le Tacon, François; Marsit, Souhir; Ropars, Jeanne; Richard, Franck; Selosse, Marc-André

2017-06-01

In this article, we review some of the best-studied fungi used as food sources, in particular, the cheese fungi, the truffles, and the fungi used for drink fermentation such as beer, wine, and sake. We discuss their history of consumption by humans and the genomic mechanisms of adaptation during artificial selection.

Industrial relevance of thermophilic Archaea.

Science.gov (United States)

Egorova, Ksenia; Antranikian, Garabed

2005-12-01

The dramatic increase of newly isolated extremophilic microorganisms, analysis of their genomes and investigations of their enzymes by academic and industrial laboratories demonstrate the great potential of extremophiles in industrial (white) biotechnology. Enzymes derived from extremophiles (extremozymes) are superior to the traditional catalysts because they can perform industrial processes even under harsh conditions, under which conventional proteins are completely denatured. In particular, enzymes from thermophilic and hyperthermophilic Archaea have industrial relevance. Despite intensive investigations, our knowledge of the structure-function relationships of their enzymes is still limited. Information concerning the molecular properties of their enzymes and genes has to be obtained to be able to understand the mechanisms that are responsible for catalytic activity and stability at the boiling point of water.

Diversity, Localization, and Physiological Properties of Filamentous Microbes Belonging to Chloroflexi Subphylum I in Mesophilic and Thermophilic Methanogenic Sludge Granules

Science.gov (United States)

Yamada, Takeshi; Sekiguchi, Yuji; Imachi, Hiroyuki; Kamagata, Yoichi; Ohashi, Akiyoshi; Harada, Hideki

2005-01-01

We previously reported that the thermophilic filamentous anaerobe Anaerolinea thermophila, which is the first cultured representative of subphylum I of the bacterial phylum Chloroflexi, not only was one of the predominant constituents of thermophilic sludge granules but also was a causative agent of filamentous sludge bulking in a thermophilic (55°C) upflow anaerobic sludge blanket (UASB) reactor in which high-strength organic wastewater was treated (Y. Sekiguchi, H. Takahashi, Y. Kamagata, A. Ohashi, and H. Harada, Appl. Environ. Microbiol. 67:5740-5749, 2001). To further elucidate the ecology and function of Anaerolinea-type filamentous microbes in UASB sludge granules, we surveyed the diversity, distribution, and physiological properties of Chloroflexi subphylum I microbes residing in UASB granules. Five different types of mesophilic and thermophilic UASB sludge were used to analyze the Chloroflexi subphylum I populations. 16S rRNA gene cloning-based analyses using a 16S rRNA gene-targeted Chloroflexi-specific PCR primer set revealed that all clonal sequences were affiliated with the Chloroflexi subphylum I group and that a number of different phylotypes were present in each clone library, suggesting the ubiquity and vast genetic diversity of these populations in UASB sludge granules. Subsequent fluorescence in situ hybridization (FISH) of the three different types of mesophilic sludge granules using a Chloroflexi-specific probe suggested that all probe-reactive cells had a filamentous morphology and were widely distributed within the sludge granules. The FISH observations also indicated that the Chloroflexi subphylum I bacteria were not always the predominant populations within mesophilic sludge granules, in contrast to thermophilic sludge granules. We isolated two mesophilic strains and one thermophilic strain belonging to the Chloroflexi subphylum I group. The physiological properties of these isolates suggested that these populations may contribute to the

An efficient Azorean thermophilic consortium for lignocellulosic biomass degradation

OpenAIRE

Martins, Rita; Teixeira, Mário; Toubarro, Duarte; Simões, Nelson; Domingues, Lucília; Teixeira, J. A.

2015-01-01

[Excerpt] Lignocellulosic plant biomass is being envisioned by biorefinery industry as an alternative to current petroleum platform because of the large scale availability, low cost and environmentally benign production. The industrial bioprocessing designed to transform lignocellulosic biomass into biofuels are harsh and the enzymatic reactions may be severely compromised reducing the production of fermentable sugars from lignocellulosic biomass. Thermophilic bacteria consortium are a potent...

Space agriculture for habitation on Mars with hyper-thermophilic aerobic composting bacteria

Science.gov (United States)

Kanazawa, S.; Ishikawa, Y.; Tomita-Yokotani, K.; Hashimoto, H.; Kitaya, Y.; Yamashita, M.; Nagatomo, M.; Oshima, T.; Wada, H.; Space Agriculture Task Force, J.

Manned Mars exploration requires recycle of materials to support human life A conceptual design is developed for space agriculture which is driven by the biologically regenerative function Hyper-thermophilic aerobic composting bacterial ecology is the core of materials recycling system to process human metabolic waste and inedible biomass and convert them to fertilizer for plants cultivation A photosynthetic reaction of plants will be driven by solar energy Water will be recycled by cultivation of plants and passing it through plant bodies Sub-surface water and atmospheric carbon dioxide are the natural resource available on Mars and these resources will be converted to oxygen and foods We envision that the agricultural system will be scaled up by importing materials from Martian environment Excess oxygen will be obtained from growing trees for structural and other components Minor elements including N P K and other traces will be introduced as fertilizers or nutrients into the agricultural materials circulation Nitrogen will be collected from Martian atmosphere We will assess biological fixation of nitrogen using micro-organisms responsible in Earth biosphere Hyper-thermophilic aerobic bacterial ecology is effective to convert waste materials into useful forms to plants This microbial technology has been well established on ground for processing sewage and waste materials For instance the hyper-thermophilic bacterial system is applied to a composting machine in a size of a trash box in home kitchen Since such a home electronics

Electricity production and microbial characterization of thermophilic microbial fuel cells.

Science.gov (United States)

Dai, Kun; Wen, Jun-Li; Zhang, Fang; Ma, Xi-Wen; Cui, Xiang-Yu; Zhang, Qi; Zhao, Ting-Jia; Zeng, Raymond J

2017-11-01

Thermophilic microbial fuel cell (TMFC) offers many benefits, but the investigations on the diversity of exoelectrogenic bacteria are scarce. In this study, a two-chamber TMFC was constructed using ethanol as an electron donor, and the microbial dynamics were analyzed by high-throughput sequencing and 16S rRNA clone-library sequencing. The open-circuit potential of TMFC was approximately 650mV, while the maximum voltage was around 550mV. The maximum power density was 437mW/m 2 , and the columbic efficiency in this work was 20.5±6.0%. The Firmicutes bacteria, related to the uncultured bacterium clone A55_D21_H_B_C01 with a similarity of 99%, accounted for 90.9% of all bacteria in the TMFC biofilm. This unknown bacterium has the potential to become a new thermophilic exoelectrogenic bacterium that is yet to be cultured. The development of TMFC-involved biotechnologies will be beneficial for the production of valuable chemicals and generation of energy in the future. Copyright © 2017 Elsevier Ltd. All rights reserved.

Bio-degradation of oily food waste employing thermophilic bacterial strains.

Science.gov (United States)

Awasthi, Mukesh Kumar; Selvam, Ammaiyappan; Chan, Man Ting; Wong, Jonathan W C

2018-01-01

The objective of this work was to isolate a novel thermophilic bacterial strain and develop a bacterial consortium (BC) for efficient degradation oily food waste. Four treatments were designed: 1:1 mixture of pre-consumption food wastes (PrCFWs) and post-consumption food wastes (PCFWs) (T-1), 1:2 mixture of PrCFWs and PCFWs mixture (T-2), PrCFWs (T-3) and PCFWs (T-4). Equal quantity of BC was inoculated into each treatment to compare the oil degradation efficiency. Results showed that after 15days of incubation, a maximum oil reduction of 65.12±0.08% was observed in treatment T-4, followed by T-2 (55.44±0.12%), T-3 (54.79±0.04%) and T-1 (52.52±0.02%), while oil reduction was negligible in control. Results indicate that the development of oil utilizing thermophilic BC was more cost-effective in solving the degradation of oily food wastes and conversion into a stable end product. Copyright © 2017 Elsevier Ltd. All rights reserved.

Thermophilic and unusually acidophilic amylase produced by a thermophilic acidophilic bacillus sp

Energy Technology Data Exchange (ETDEWEB)

Uchino, F

1982-01-01

Bacillus sp. 11-1S, a thermophilic acidophilic bacterial strain, produced an extracellular amylase with unusual characteristics. The enzyme was purified 40-fold by SE-Sephadex column chromatography. The pH optimum for activity was 2.0, and substantial activity was noted in the pH range of 1.5-3.5. The optimal temperature was 70 degrees C, but the activity decreased markedly in lower reaction temperatures. Arrhenius plots of the reaction showed two straight lines intersecting at about 50 degrees C. The activity or stability of the enzyme was not likely to depend on Ca2+. The molecular weight of the enzyme was 54,000 calculated from the electrophoretic mobility. The enzyme behaved like an alpha-amylase (1,4-alpha-D- glucan glucanohydrolase, E.C. 3.2.1.1). About 34% of glucosidic linkages of soluble starch was hydrolyzed at 65 degrees C and pH 2.0, in 24 hours, and the major products were maltotriose and maltose. (Refs. 14).

Adsorption of cellulase on cellulolytic enzyme lignin from lodgepole pine.

Science.gov (United States)

Tu, Maobing; Pan, Xuejun; Saddler, Jack N

2009-09-09

Enzymatic hydrolysis of lignocellulosic materials is significantly affected by cellulase adsorption onto the lignocellulosic substrates and lignin. The presence of lignin plays an important role in lignocellulosic hydrolysis and enzyme recycling. Three cellulase preparations (Celluclast, Spezyme CP, and MSUBC) were evaluated to determine their adsorption onto cellulolytic enzyme lignin (CEL) from steam-exploded Lodgepole pine (SELP) and ethanol (organosolv)-pretreated Lodgepole pine (EPLP). The adsorption affinity of cellulase (Celluclast) onto isolated lignin (CEL-EPLP and CEL-SELP) was slightly higher than that from corresponding EPLP and SELP substrates on the basis of the Langmuir constants. Effects of temperature, ionic strength, and surfactant on cellulase adsorption onto isolated lignin were also explored in this study. Thermodynamic analysis of enzyme adsorption onto isolated lignin (Gibbs free energy change DeltaG(0) approximately -30 kJ/mol) indicated this adsorption was a spontaneous process. The addition of surfactant (0.2% w/v) could reduce the adsorption of cellulase onto CEL-SELP by 60%. Two types of adsorption isotherm were compared for cellulase adsorption onto isolated lignin. A Langmuir adsorption isotherm showed better fit for the experimental data than a Freundlich adsorption isotherm.

Rock phosphate solubilizing and cellulolytic actinomycete isolates of earthworm casts

Science.gov (United States)

Mba, Caroline C.

1994-03-01

Four microbial isolates, OP2, OP3, OP6, and OP7, of earthworm casts of Pontoscolex corethrurus were found to be acid tolerant actinomycetes and efficient rock phosphate (RP) solubilizers that could grow fast on NH4Cl-enriched or N-free carboxymethyl cellulose or glucose as sole carbon source. CMC (carboxymethyl cellulose) induced production of extracellular cellulase enzyme and the production of reducing sugar in all the isolates. RP solubilizing power was observed to be inversely related to glucose consumption. The most efficient RP solubilizer was found to consume the least glucose. Growth was faster on cellulose than on glucose media. N-free CMC induced greater glucose production than NH4Cl-enriched CMC medium. Both CMC and glucose media were acidified by all the isolates, however, RP solubilizing power decreased with acidification. Solubilization power was greatest with isolate OP7, which also produced the greatest amount of reducing sugar per gram CMC. Both RP solubilizing power and the cellulolytic efficiency varied among isolates. A minimum of 631 µg P/0.1 g RP and a maximum of 951.4 µg P/0.1 g RP was recorded.

Fungi

DEFF Research Database (Denmark)

Hajek, Ann E.; Meyling, Nicolai Vitt

2018-01-01

been the focus of most ecological research. Some taxa of invertebrate pathogenic fungi have evolved adaptations for utilizing living plants as substrates, and these lifestyles have recently received increased attention from researchers following the initial documentations of such plant associations...

Higher marine fungi from mangroves (Manglicolous fungi)

Digital Repository Service at National Institute of Oceanography (India)

ChinnaRaj, S.

of higher marine fungi which included 23 Ascomycetes, 2 Basidiomycetes and 17 Deuteromycetes (Kohlmeyer and Kohlmeyer, 1979). Hyde (1990a) listed 120 species from 29 mangroves from all over the World this includes 87 Ascomycetes, 2 Basidiomycetes and 31...

Hydrogen and methane production from desugared molasses using a two‐stage thermophilic anaerobic process

DEFF Research Database (Denmark)

Kongjan, Prawit; O-Thong, Sompong; Angelidaki, Irini

2013-01-01

Hydrogen and methane production from desugared molasses by a two‐stage thermophilic anaerobic process was investigated in a series of two up‐flow anaerobic sludge blanket (UASB) reactors. The first reactor that was dominated with hydrogen‐producing bacteria of Thermoanaerobacterium thermosaccharo......Hydrogen and methane production from desugared molasses by a two‐stage thermophilic anaerobic process was investigated in a series of two up‐flow anaerobic sludge blanket (UASB) reactors. The first reactor that was dominated with hydrogen‐producing bacteria of Thermoanaerobacterium...... molasses. Furthermore, the mixed gas with a volumetric content of 16.5% H2, 38.7% CO2, and 44.8% CH4, containing approximately 15% energy by hydrogen is viable to be bio‐hythane....

Gelria glutamica gen. nov., sp. a thermophilic oligately syntrophic glutamate-degrading anaerobe

NARCIS (Netherlands)

Plugge, C.M.; Balk, M.; Zoetendal, E.G.; Stams, A.J.M.

2002-01-01

A novel anaerobic, Gram-positive, thermophilic, spore-forming, obligately syntrophic, glutamate-degrading bacterium, strain TGO(T), was isolated from a propionate-oxidizing methanogenic enrichment culture. The axenic culture was obtained by growing the bacterium on pyruvate. Cells were rod-shaped

Biogas Upgrading via Hydrogenotrophic Methanogenesis in Two-Stage Continuous Stirred Tank Reactors at Mesophilic and Thermophilic Conditions

DEFF Research Database (Denmark)

Bassani, Ilaria; Kougias, Panagiotis; Treu, Laura

2015-01-01

This study proposes an innovative setup composed by two stage reactors to achieve biogas upgrading coupling the CO2 in the biogas with external H2 and subsequent conversion into CH4 by hydrogenotrophic methanogenesis. In this configuration, the biogas produced in the first reactor was transferred...... production and CO2 conversion was recorded. The consequent increase of pH did not inhibit the process indicating adaptation of microorganisms to higher pH levels. The effects of H2 on the microbial community were studied using high-throughput Illumina random sequences and full-length 16S rRNA genes extracted...... to the second one, where H2 was injected. This configuration was tested at both mesophilic and thermophilic conditions. After H2 addition, the produced biogas was upgraded to average CH4 content of 89% in the mesophilic reactor and 85% in the thermophilic. At thermophilic conditions, a higher efficiency of CH4...

Community phylogenetic analysis of moderately thermophilic cyanobacterial mats from China, the Philippines and Thailand.

Science.gov (United States)

Hongmei, Jing; Aitchison, Jonathan C; Lacap, Donnabella C; Peerapornpisal, Yuwadee; Sompong, Udomluk; Pointing, Stephen B

2005-08-01

Most community molecular studies of thermophilic cyanobacterial mats to date have focused on Synechococcus occurring at temperatures of approximately 50-65 degrees C. These reveal that molecular diversity exceeds that indicated by morphology, and that phylogeographic lineages exist. The moderately thermophilic and generally filamentous cyanobacterial mat communities occurring at lower temperatures have not previously been investigated at the community molecular level. Here we report community diversity in mats of 42-53 degrees C recovered from previously unstudied geothermal locations. Separation of 16S rRNA gene-defined genotypes from community DNA was achieved by DGGE. Genotypic diversity was greater than morphotype diversity in all mats sampled, although genotypes generally corresponded to observed morphotypes. Thirty-six sequences were recovered from DGGE bands. Phylogenetic analyses revealed these to form novel thermophilic lineages distinct from their mesophilic counterparts, within Calothrix, Cyanothece, Fischerella, Phormidium, Pleurocapsa, Oscillatoria and Synechococcus. Where filamentous cyanobacterial sequences belonging to the same genus were recovered from the same site, these were generally closely affiliated. Location-specific sequences were observed for some genotypes recovered from geochemically similar yet spatially separated sites, thus providing evidence for phylogeographic lineages that evolve in isolation. Other genotypes were more closely affiliated to geographically remote counterparts from similar habitats suggesting that adaptation to certain niches is also important.

Enumeration of fungi in barley

CSIR Research Space (South Africa)

Rabie, CJ

1997-04-01

Full Text Available Estimation of fungal contamination of barley grain is important as certain fungi can proliferate during the malting process. The following factors which may affect the enumeration of fungi were evaluated: dilution versus direct plating, pre...

Molecular and phenotypic characterization of anamorphic fungi

OpenAIRE

Madrid Lorca, Hugo

2011-01-01

Anamorphic fungi (those reproducing asexually) are a big part of kingdom Fungi. Most of them occur as saprobes in nature, but numerous species are pathogenic to plants and animals including man. With the aim of contributing to the knowledge of the diversity and distribution of anamorphic fungi, we performed a phenotypic and molecular characterization of environmental and clinical isolates of these fungi. Based on a polyphasic taxonomy approach which included morphology, physiology and DNA seq...

Some mycogenous fungi from Poland

Directory of Open Access Journals (Sweden)

Andrzej Chlebicki

2014-08-01

Full Text Available In the present paper the results of earlier studies on mycogenous fungi which were gathered occasionally are summarized. Fifieen specres. previously Pyrenomycetes s.l., have been found growing on other fungi Immothia hypoxylon and Lophiostoma polyporicola are new species to the Polish mycoflora. Sphaeronaemella Kulczyńskiana described by K. R o u p p e r t (1912 is considered to be Eleuteromyces subultus. Relatively high number of fungi inhabiting stromata of Diatrypella favacea is probably connected with its early colonization of the Polish area.

Recombinant HAP Phytase of the Thermophilic Mold Sporotrichum thermophile: Expression of the Codon-Optimized Phytase Gene in Pichia pastoris and Applications.

Science.gov (United States)

Ranjan, Bibhuti; Satyanarayana, T

2016-02-01

The codon-optimized phytase gene of the thermophilic mold Sporotrichum thermophile (St-Phy) was expressed in Pichia pastoris. The recombinant P. pastoris harboring the phytase gene (rSt-Phy) yielded a high titer of extracellular phytase (480 ± 23 U/mL) on induction with methanol. The recombinant phytase production was ~40-fold higher than that of the native fungal strain. The purified recombinant phytase (rSt-Phy) has the molecular mass of 70 kDa on SDS-PAGE, with K m and V max (calcium phytate), k cat and k cat/K m values of 0.147 mM and 183 nmol/mg s, 1.3 × 10(3)/s and 8.84 × 10(6)/M s, respectively. Mg(2+) and Ba(2+) display a slight stimulatory effect, while other cations tested exert inhibitory action on phytase. The enzyme is inhibited by chaotropic agents (guanidinium hydrochloride, potassium iodide, and urea), Woodward's reagent K and 2,3-bunatedione, but resistant to both pepsin and trypsin. The rSt-Phy is useful in the dephytinization of broiler feeds efficiently in simulated gut conditions of chick leading to the liberation of soluble inorganic phosphate with concomitant mitigation in antinutrient effects of phytates. The addition of vanadate makes it a potential candidate for generating haloperoxidase, which has several applications.

Thermophilic and alkaliphilic Actinobacteria: biology and potential applications

Science.gov (United States)

Shivlata, L.; Satyanarayana, Tulasi

2015-01-01

Microbes belonging to the phylum Actinobacteria are prolific sources of antibiotics, clinically useful bioactive compounds and industrially important enzymes. The focus of the current review is on the diversity and potential applications of thermophilic and alkaliphilic actinobacteria, which are highly diverse in their taxonomy and morphology with a variety of adaptations for surviving and thriving in hostile environments. The specific metabolic pathways in these actinobacteria are activated for elaborating pharmaceutically, agriculturally, and biotechnologically relevant biomolecules/bioactive compounds, which find multifarious applications. PMID:26441937

Production of 2-deoxyribose 5-phosphate from fructose to demonstrate a potential of artificial bio-synthetic pathway using thermophilic enzymes.

Science.gov (United States)

Honda, Kohsuke; Maya, Shohei; Omasa, Takeshi; Hirota, Ryuichi; Kuroda, Akio; Ohtake, Hisao

2010-08-02

Six thermophilic enzymes from Thermus thermophilus were used to construct an 'artificial bio-synthetic pathway' for the production of 2-deoxyribose 5-phosphate from fructose. By a simple operation using six recombinant Escherichia coli strains producing the thermophilic enzymes, respectively, fructose was converted to 2-deoxyribose 5-phosphate with a molar yield of 55%. Copyright 2010 Elsevier B.V. All rights reserved.

Substrate and product inhibition of hydrogen production by the extreme thermophile, Caldicellulosiruptor saccharolyticus

NARCIS (Netherlands)

Niel, van E.W.J.; Claassen, P.A.M.; Stams, A.J.M.

2003-01-01

Substrate and product inhibition of hydrogen production during sucrose fermentation by the extremely thermophilic bacterium Caldicellulosiruptor saccharolyticus was studied. The inhibition kinetics were analyzed with a noncompetitive, nonlinear inhibition model. Hydrogen was the most severe

Advances in Genomics of Entomopathogenic Fungi.

Science.gov (United States)

Wang, J B; St Leger, R J; Wang, C

2016-01-01

Fungi are the commonest pathogens of insects and crucial regulators of insect populations. The rapid advance of genome technologies has revolutionized our understanding of entomopathogenic fungi with multiple Metarhizium spp. sequenced, as well as Beauveria bassiana, Cordyceps militaris, and Ophiocordyceps sinensis among others. Phylogenomic analysis suggests that the ancestors of many of these fungi were plant endophytes or pathogens, with entomopathogenicity being an acquired characteristic. These fungi now occupy a wide range of habitats and hosts, and their genomes have provided a wealth of information on the evolution of virulence-related characteristics, as well as the protein families and genomic structure associated with ecological and econutritional heterogeneity, genome evolution, and host range diversification. In particular, their evolutionary transition from plant pathogens or endophytes to insect pathogens provides a novel perspective on how new functional mechanisms important for host switching and virulence are acquired. Importantly, genomic resources have helped make entomopathogenic fungi ideal model systems for answering basic questions in parasitology, entomology, and speciation. At the same time, identifying the selective forces that act upon entomopathogen fitness traits could underpin both the development of new mycoinsecticides and further our understanding of the natural roles of these fungi in nature. These roles frequently include mutualistic relationships with plants. Genomics has also facilitated the rapid identification of genes encoding biologically useful molecules, with implications for the development of pharmaceuticals and the use of these fungi as bioreactors. Copyright © 2016 Elsevier Inc. All rights reserved.

Structural insights into cellulolytic and chitinolytic enzymes revealing crucial residues of insect β-N-acetyl-D-hexosaminidase.

Directory of Open Access Journals (Sweden)

Tian Liu

Full Text Available The chemical similarity of cellulose and chitin supports the idea that their corresponding hydrolytic enzymes would bind β-1,4-linked glucose residues in a similar manner. A structural and mutational analysis was performed for the plant cellulolytic enzyme BGlu1 from Oryza sativa and the insect chitinolytic enzyme OfHex1 from Ostrinia furnacalis. Although BGlu1 shows little amino-acid sequence or topological similarity with OfHex1, three residues (Trp(490, Glu(328, Val(327 in OfHex1, and Trp(358, Tyr(131 and Ile(179 in BGlu1 were identified as being conserved in the +1 sugar binding site. OfHex1 Glu(328 together with Trp(490 was confirmed to be necessary for substrate binding. The mutant E328A exhibited a 8-fold increment in K(m for (GlcNAc(2 and a 42-fold increment in K(i for TMG-chitotriomycin. A crystal structure of E328A in complex with TMG-chitotriomycin was resolved at 2.5 Å, revealing the obvious conformational changes of the catalytic residues (Glu(368 and Asp(367 and the absence of the hydrogen bond between E328A and the C3-OH of the +1 sugar. V327G exhibited the same activity as the wild-type, but acquired the ability to efficiently hydrolyse β-1,2-linked GlcNAc in contrast to the wild-type. Thus, Glu(328 and Val(327 were identified as important for substrate-binding and as glycosidic-bond determinants. A structure-based sequence alignment confirmed the spatial conservation of these three residues in most plant cellulolytic, insect and bacterial chitinolytic enzymes.

Thermophilic methanogenic Archaea in compost material: occurrence, persistence and possible mechanisms for their distribution to other environments.

Science.gov (United States)

Thummes, Kathrin; Schäfer, Jenny; Kämpfer, Peter; Jäckel, Udo

2007-12-01

Since compost is widely used as soil amendment and the fact that during the processing of compost material high amounts of microorganisms are released into the air, we investigated whether compost may act as a carrier for thermophilic methanogens to temperate soils. All eight investigated compost materials showed a clear methane production potential between 0.01 and 0.98 micromol CH(4) g dw(-1)h(-1) at 50 degrees C. Single strand conformation polymorphism (SSCP) and cloning analysis indicated the presence of Methanosarcina thermophila, Methanoculleus thermophilus, and Methanobacterium formicicum. Bioaerosols collected during the turning of a compost pile showed both a highly similar SSCP profile compared to the corresponding compost material and clear methane production during anoxic incubation in selective medium at 50 degrees C. Both observations indicated a considerable release of thermophilic methanogens into the air. To analyse the persistence of compost-borne thermophilic methanogens in temperate oxic soils, we therefore studied their potential activity in compost and compost/soil mixtures, which was brought to a meadow soil, as well as in an agricultural soil fertilised with compost. After 24h anoxic incubation at 50 degrees C, all samples containing compost showed a clear methanogenic activity, even 1 year after application. In combination with the in vitro observed resilience of the compost-borne methanogens against desiccation and UV radiation we assume that compost material acts as an effective carrier for the distribution of thermophilic methanogens by fertilisation and wind.

A Combination of Stable Isotope Probing, Illumina Sequencing, and Co-occurrence Network to Investigate Thermophilic Acetate- and Lactate-Utilizing Bacteria.

Science.gov (United States)

Sun, Weimin; Krumins, Valdis; Dong, Yiran; Gao, Pin; Ma, Chunyan; Hu, Min; Li, Baoqin; Xia, Bingqing; He, Zijun; Xiong, Shangling

2018-01-01

Anaerobic digestion is a complicated microbiological process that involves a wide diversity of microorganisms. Acetate is one of the most important intermediates, and interactions between acetate-oxidizing bacteria and archaea could play an important role in the formation of methane in anoxic environments. Anaerobic digestion at thermophilic temperatures is known to increase methane production, but the effects on the microbial community are largely unknown. In the current study, stable isotope probing was used to characterize acetate- and lactate-oxidizing bacteria in thermophilic anaerobic digestion. In microcosms fed 13 C-acetate, bacteria related to members of Clostridium, Hydrogenophaga, Fervidobacterium, Spirochaeta, Limnohabitans, and Rhodococcus demonstrated elevated abundances of 13 C-DNA fractions, suggesting their activities in acetate oxidation. In the treatments fed 13 C-lactate, Anaeromyxobacter, Desulfobulbus, Syntrophus, Cystobacterineae, and Azospira were found to be the potential thermophilic lactate utilizers. PICRUSt predicted that enzymes related to nitrate and nitrite reduction would be enriched in 13 C-DNA fractions, suggesting that the acetate and lactate oxidation may be coupled with nitrate and/or nitrite reduction. Co-occurrence network analysis indicated bacterial taxa not enriched in 13 C-DNA fractions that may also play a critical role in thermophilic anaerobic digestion.

Genetically Engineering Entomopathogenic Fungi.

Science.gov (United States)

Zhao, H; Lovett, B; Fang, W

2016-01-01

Entomopathogenic fungi have been developed as environmentally friendly alternatives to chemical insecticides in biocontrol programs for agricultural pests and vectors of disease. However, mycoinsecticides currently have a small market share due to low virulence and inconsistencies in their performance. Genetic engineering has made it possible to significantly improve the virulence of fungi and their tolerance to adverse conditions. Virulence enhancement has been achieved by engineering fungi to express insect proteins and insecticidal proteins/peptides from insect predators and other insect pathogens, or by overexpressing the pathogen's own genes. Importantly, protein engineering can be used to mix and match functional domains from diverse genes sourced from entomopathogenic fungi and other organisms, producing insecticidal proteins with novel characteristics. Fungal tolerance to abiotic stresses, especially UV radiation, has been greatly improved by introducing into entomopathogens a photoreactivation system from an archaean and pigment synthesis pathways from nonentomopathogenic fungi. Conversely, gene knockout strategies have produced strains with reduced ecological fitness as recipients for genetic engineering to improve virulence; the resulting strains are hypervirulent, but will not persist in the environment. Coupled with their natural insect specificity, safety concerns can also be mitigated by using safe effector proteins with selection marker genes removed after transformation. With the increasing public concern over the continued use of synthetic chemical insecticides and growing public acceptance of genetically modified organisms, new types of biological insecticides produced by genetic engineering offer a range of environmentally friendly options for cost-effective control of insect pests. Copyright © 2016 Elsevier Inc. All rights reserved.

Fungi isolated in school buildings

Directory of Open Access Journals (Sweden)

Elżbieta Ejdys

2013-12-01

Full Text Available The aim of the study was to determine the species composition of fungi occurring on wall surfaces and in the air in school buildings. Fungi isolated from the air using the sedimentation method and from the walls using the surface swab technique constituted the study material. Types of finish materials on wall surfaces were identified and used in the analysis. Samples were collected in selected areas in two schools: classrooms, corridors, men's toilets and women's toilets, cloakrooms, sports changing rooms and shower. Examinations were conducted in May 2005 after the heating season was over. Fungi were incubated on Czapek-Dox medium at three parallel temperatures: 25, 37 and 40°C, for at least three weeks. A total of 379 isolates of fungi belonging to 32 genera of moulds, yeasts and yeast-like fungi were obtained from 321 samples in the school environment. The following genera were isolated most frequently: Aspergillus, Penicillium and Cladosporium. Of the 72 determined species, Cladosporium herbarum, Aspergillus fumigatus and Penicillium chrysogenum occurred most frequently in the school buildings. Wall surfaces were characterised by an increased prevalence of mycobiota in comparison with the air in the buildings, with a slightly greater species diversity. A certain species specificity for rough and smooth wall surfaces was demonstrated. Fungi of the genera Cladosporium and Emericella with large spores adhered better to smooth surfaces while those of the genus Aspergillus with smaller conidia adhered better to rough surfaces. The application of three incubation temperatures helped provide a fuller picture of the mycobiota in the school environment.

Cloning, expression, crystallization and preliminary X-ray characterization of cytochrome c552 from a moderate thermophilic bacterium, Hydrogenophilus thermoluteolus

International Nuclear Information System (INIS)

Ichiki, Shin-ichi; Nakamura, Shota; Ohkubo, Tadayasu; Kobayashi, Yuji; Hasegawa, Jun; Uchiyama, Susumu; Nishihara, Hirofumi; Mizuta, Keiko; Sambongi, Yoshihiro

2005-01-01

Cytochrome c 552 of a moderate thermophile, H. thermoluteolus, was overexpressed in E. coli and crystallized for X-ray diffraction study. The amino-acid sequence of cytochrome c 552 (PH c 552 ) from a moderately thermophilic bacterium, Hydrogenophilus thermoluteolus, was more than 50% identical to that of cytochrome c from an extreme thermophile, Hydrogenobacter thermophilus (HT c 552 ), and from a mesophile, Pseudomonas aeruginosa (PA c 551 ). The PH c 552 gene was overexpressed as a correctly processed holoprotein in the Escherichia coli periplasm. The overexpressed PH c 552 has been crystallized by vapour diffusion from polyethylene glycol 4000 pH 6.5. The crystals belong to space group C222 1 , with unit-cell parameters a = 48.98, b = 57.99, c = 56.20 Å. The crystals diffract X-rays to around 2.1 Å resolution

Selection of micro-organisms and fermentation conditions of oil palm empty fruit bunch (EFB) and palm press fibre (PPF)

International Nuclear Information System (INIS)

Mat Rasol Awang; Hassan Hamdani Mutaat; Tamikazu Kume; Hitoshi Ito

1998-01-01

The selection of useful microorganisms was made by trial cultivation of various cellulolytic fungi on EFB and PPF. Several fermentation conditions were performed involving adjusting alkali treatment conditions, pH, changing media composition and preparation technique of solid culture media. Basic the preparation of the solid culture media was made by dissolving inorganic salts together with micro-nutrients and then added to the alkali treated EFB and PPF. In the cultivation of mushrooms, the preparation of solid culture media was adopted from mushroom growers technique. The criteria of a good degradation ability of fungi were evaluated based on the percentage of crude fibre degradation of EFB and PPF by fungi. The nutritional values of the products such as protein was also characterised

Genomic insights into the fungal lignocellulolytic system of Myceliophthora thermophila

Directory of Open Access Journals (Sweden)

Anthi eKarnaouri

2014-06-01

Full Text Available The microbial conversion of solid cellulosic biomass to liquid biofuels may provide a renewable energy source for transportation fuels. Cellulolytic fungi represent a promising group of organisms, as they have evolved complex systems for adaptation to their natural habitat. The filamentous fungus Myceliophthora thermophila constitutes an exceptionally powerful cellulolytic microorganism that synthesizes a complete set of enzymes necessary for the breakdown of plant cell wall. The genome of this fungus has been recently sequenced and annotated, allowing systematic examination and identification of enzymes required for the degradation of lignocellulosic biomass. The genomic analysis revealed the existence of an expanded enzymatic repertoire including numerous cellulases, hemicellulases and enzymes with auxiliary activities, covering the most of the recognized CAZy families. Most of them were predicted to possess a secretion signal and undergo through post translational glycosylation modifications. These data offer a better understanding of activities embedded in fungal lignocellulose decomposition mechanisms and suggest that M. thermophila could be made usable as an industrial production host for cellulolytic and hemicellulolytic enzymes.

Thermophilic and alkaliphilic Actinobacteria: Biology and potential applications

Directory of Open Access Journals (Sweden)

L eShivlata

2015-09-01

Full Text Available Microbes belonging to the phylum Actinobacteria are prolific sources of antibiotics, clinically useful bioactive compounds and industrially important enzymes. The focus of the current review is on the diversity and potential applications of thermophilic and alkaliphilic actinobacteria, which are highly diverse in their taxonomy and morphology with a variety of adaptations for surviving and thriving in hostile environments. The specific metabolic pathways in these actinobacteria are activated for elaborating pharmaceutically, agriculturally and biotechnologically relevant biomolecules/bioactive compounds, which find multifarious applications.

Comprehensive microbial analysis of combined mesophilic anaerobic-thermophilic aerobic process treating high-strength food wastewater.

Science.gov (United States)

Jang, Hyun Min; Ha, Jeong Hyub; Park, Jong Moon; Kim, Mi-Sun; Sommer, Sven G

2015-04-15

A combined mesophilic anaerobic-thermophilic aerobic process was used to treat high-strength food wastewater in this study. During the experimental period, most of solid residue from the mesophilic anaerobic reactor (R1) was separated by centrifugation and introduced into the thermophilic aerobic reactor (R2) for further digestion. Then, thermophilic aerobically-digested sludge was reintroduced into R1 to enhance reactor performance. The combined process was operated with two different Runs: Run I with hydraulic retention time (HRT) = 40 d (corresponding OLR = 3.5 kg COD/m(3) d) and Run II with HRT = 20 d (corresponding OLR = 7 kg COD/m(3)). For a comparison, a single-stage mesophilic anaerobic reactor (R3) was operated concurrently with same OLRs and HRTs as the combined process. During the overall digestion, all reactors showed high stability without pH control. The combined process demonstrated significantly higher organic matter removal efficiencies (over 90%) of TS, VS and COD and methane production than did R3. Quantitative real-time PCR (qPCR) results indicated that higher populations of both bacteria and archaea were maintained in R1 than in R3. Pyrosequencing analysis revealed relatively high abundance of phylum Actinobacteria in both R1 and R2, and a predominance of phyla Synergistetes and Firmicutes in R3 during Run II. Furthermore, R1 and R2 shared genera (Prevotella, Aminobacterium, Geobacillus and Unclassified Actinobacteria), which suggests synergy between mesophilic anaerobic digestion and thermophilic aerobic digestion. For archaea, in R1 methanogenic archaea shifted from genus Methanosaeta to Methanosarcina, whereas genera Methanosaeta, Methanobacterium and Methanoculleus were predominant in R3. The results demonstrated dynamics of key microbial populations that were highly consistent with an enhanced reactor performance of the combined process. Copyright © 2015 Elsevier Ltd. All rights reserved.

Molecular studies on protein- and carbohydrate-converting ezymes from thermophilic bacteria

NARCIS (Netherlands)

Kluskens, L.D.

2004-01-01

Microorganisms that are able to grow at hightemperatures are calledthermophiles(>55

Reactivity improvement of cellulolytic enzyme lignin via mild hydrothermal modification.

Science.gov (United States)

Ma, Zhuoming; Tang, Jiafa; Li, Shujun; Suo, Enxiang

2017-12-01

Isolated by the cellulolytic enzyme lignin (CEL) process, water-alcohol (1:1, v/v) was introduced as co-solvent in the process of the hydrothermal treatment. The modification parameters such as reaction temperature and time, solid-to-liquid ratio, and catalysts (NaOH and NaOAlO 2 ) have been investigated in terms of the specific lignin properties, such as the phenolic hydroxyl content (OH phen ), DPPH free radical scavenging rate, and formaldehyde value. The CELs were also characterized by GPC, FT-IR and 1 H NMR spectroscopy, and Py-GC/MS. The key data are under optimal lignin modification conditions (solid-to-liquid ratio of 1:10 (w/v) and a temperature of 250°C for 60min) are: OH phen content: 2.50mmol/g; half maximal inhibitory concentration (IC 50 ) towards DPPH free radicals: 88.2mg/L; formaldehyde value: 446.9g/kg). Both base catalysts decrease the residue rate, but phenol reactivities of the products were also detracted. Py-GC/MS results revealed that modified lignin had a higher phenolic composition than the CEL did, especially the modified lignin without catalyst (ML), which represented 74.51% phenolic content. Copyright © 2017. Published by Elsevier Inc.

Survival of thermophilic and hyperthermophilic microorganisms after exposure to UV-C, ionizing radiation and desiccation.

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Beblo, Kristina; Douki, Thierry; Schmalz, Gottfried; Rachel, Reinhard; Wirth, Reinhard; Huber, Harald; Reitz, Günther; Rettberg, Petra

2011-11-01

In this study, we investigated the ability of several (hyper-) thermophilic Archaea and phylogenetically deep-branching thermophilic Bacteria to survive high fluences of monochromatic UV-C (254 nm) and high doses of ionizing radiation, respectively. Nine out of fourteen tested microorganisms showed a surprisingly high tolerance against ionizing radiation, and two species (Aquifex pyrophilus and Ignicoccus hospitalis) were even able to survive 20 kGy. Therefore, these species had a comparable survivability after exposure to ionizing radiation such as Deinococcus radiodurans. In contrast, there was nearly no difference in survival of the tested strains after exposure to UV-C under anoxic conditions. If the cells had been dried in advance of UV-C irradiation, they were more sensitive to UV-C radiation compared with cells irradiated in liquid suspension; this effect could be reversed by the addition of protective material like sulfidic ores before irradiation. By exposure to UV-C, photoproducts were formed in the DNA of irradiated Archaea and Bacteria. The distribution of the main photoproducts was species specific, but the amount of the photoproducts was only partly dependent on the applied fluence. Overall, our results show that tolerance to radiation seems to be a common phenomenon among thermophilic and hyperthermophilic microorganisms.

Occurrence of keratinophilic fungi on Indian birds.

Science.gov (United States)

Dixit, A K; Kushwaha, R K

1991-01-01

Keratinophilic fungi were isolated from feathers of most common Indian birds, viz. domestic chicken (Gallus domesticus), domestic pigeon (Columba livia), house sparrow (Passer domesticus), house crow (Corvus splendens), duck (Anas sp.), rose-ringed parakeet (Psittacula krameri). Out of 87 birds, 58 yielded 4 keratinophilic fungal genera representing 13 fungal species and one sterile mycelium. The isolated fungi were cultured on Sabouraud's dextrose agar at 28 +/- 2 degrees C. Chrysosporium species were isolated on most of the birds. Chrysosporium lucknowense and Chrysosporium tropicum were the most common fungal species associated with these Indian birds. Maximum occurrence of fungi (47%) was recorded on domestic chickens and the least number of keratinophilic fungi was isolated from the domestic pigeon and duck. The average number of fungi per bird was found to be the 0.44.

Anaerobic digestion of whole stillage from dry-grind corn ethanol plant under mesophilic and thermophilic conditions.

Science.gov (United States)

Eskicioglu, Cigdem; Kennedy, Kevin J; Marin, Juan; Strehler, Benjamin

2011-01-01

Anaerobic digestion of whole stillage from a dry-grind corn-based ethanol plant was evaluated by batch and continuous-flow digesters under thermophilic and mesophilic conditions. At whole corn stillage concentrations of 6348 to 50,786 mg total chemical oxygen demand (TCOD)/L, at standard temperature (0 °C) and pressure (1 atm), preliminary biochemical methane potential assays produced 88±8 L (49±5 L CH4) and 96±19 L (65±14 L CH4) biogas per L stillage from mesophilic and thermophilic digesters, respectively. Continuous-flow studies for the full-strength stillage (TCOD=254 g/L) at organic loadings of 4.25, 6.30 and 9.05 g TCOD/L days indicated unstable performance for the thermophilic digester. Among the sludge retention times (SRTs) of 60, 45 and 30 days tested, the mesophilic digestion was successful only at 60 days-SRT which does not represent a practical operation time for a large scale bioethanol plant. Future laboratory studies will focus on different reactor configurations to reduce the SRT needed in the digesters. Copyright © 2010 Elsevier Ltd. All rights reserved.

Occurrence and molecular characterization of cultivable mesophilic and thermophilic obligate anaerobic bacteria isolated from paper mills.

Science.gov (United States)

Suihko, Maija-Liisa; Partanen, Laila; Mattila-Sandholm, Tiina; Raaska, Laura

2005-08-01

The aim of this work was to characterize the cultivable obligate anaerobic bacterial population in paper mill environments. A total of 177 anaerobically grown bacterial isolates were screened for aerotolerance, from which 67 obligate anaerobes were characterized by automated ribotyping and 41 were further identified by partial 16S rDNA sequencing. The mesophilic isolates indicated 11 different taxa (species) within the genus Clostridium and the thermophilic isolates four taxa within the genus Thermoanaerobacterium and one within Thermoanaerobacter (both formerly Clostridium). The most widespread mesophilic bacterium was closely related to C. magnum and occurred in three of four mills. One mill was contaminated with a novel mesophilic bacterium most closely related to C. thiosulfatireducens. The most common thermophile was T. thermosaccharolyticum, occurring in all four mills. The genetic relationships of the mill isolates to described species indicated that most of them are potential members of new species. On the basis of identical ribotypes clay could be identified to be the contamination source of thermophilic bacteria. Automated ribotyping can be a useful tool for the identification of clostridia as soon as comprehensive identification libraries are available.

Isolation and characterization of two novel ethanol-tolerant facultative-anaerobic thermophilic bacteria strains from waste compost.

Science.gov (United States)

Fong, Jiunn C N; Svenson, Charles J; Nakasugi, Kenlee; Leong, Caine T C; Bowman, John P; Chen, Betty; Glenn, Dianne R; Neilan, Brett A; Rogers, Peter L

2006-10-01

In a search for potential ethanologens, waste compost was screened for ethanol-tolerant thermophilic microorganisms. Two thermophilic bacterial strains, M5EXG and M10EXG, with tolerance of 5 and 10% (v/v) ethanol, respectively, were isolated. Both isolates are facultative anaerobic, non-spore forming, non-motile, catalase-positive, oxidase-negative, Gram-negative rods that are capable of utilizing a range of carbon sources including arabinose, galactose, mannose, glucose and xylose and produce low amounts of ethanol, acetate and lactate. Growth of both isolates was observed in fully defined minimal media within the temperature range 50-80 degrees C and pH 6.0-8.0. Phylogenetic analysis of the 16S rDNA sequences revealed that both isolates clustered with members of subgroup 5 of the genus Bacillus. G+C contents and DNA-DNA relatedness of M5EXG and M10EXG revealed that they are strains belonging to Geobacillus thermoglucosidasius. However, physiological and biochemical differences were evident when isolates M5EXG and M10EXG were compared with G. thermoglucosidasius type strain (DSM 2542(T)). The new thermophilic, ethanol-tolerant strains of G. thermoglucosidasius may be candidates for ethanol production at elevated temperatures.

Microbial community dynamics in thermophilic undefined milk starter cultures.

Science.gov (United States)

Parente, Eugenio; Guidone, Angela; Matera, Attilio; De Filippis, Francesca; Mauriello, Gianluigi; Ricciardi, Annamaria

2016-01-18

Model undefined thermophilic starter cultures were produced from raw milk of nine pasta-filata cheesemaking plants using a selective procedure based on pasteurization and incubation at high temperature with the objective of studying the microbial community dynamics and the variability in performances under repeated (7-13) reproduction cycles with backslopping. The traditional culture-dependent approach, based on random isolation and molecular characterization of isolates was coupled to the determination of pH and the evaluation of the ability to produce acid and fermentation metabolites. Moreover, a culture-independent approach based on amplicon-targeted next-generation sequencing was employed. The microbial diversity was evaluated by 16S rRNA gene sequencing (V1-V3 regions), while the microdiversity of Streptococcus thermophilus populations was explored by using novel approach based on sequencing of partial amplicons of the phosphoserine phosphatase gene (serB). In addition, the occurrence of bacteriophages was evaluated by qPCR and by multiplex PCR. Although it was relatively easy to select for a community dominated by thermophilic lactic acid bacteria (LAB) within a single reproduction cycle, final pH, LAB populations and acid production activity fluctuated over reproduction cycles. Both culture-dependent and -independent methods showed that the cultures were dominated by either S. thermophilus or Lactobacillus delbrueckii subsp. lactis or by both species. Nevertheless, subdominant mesophilic species, including lactococci and spoilage organisms, persisted at low levels. A limited number of serB sequence types (ST) were present in S. thermophilus populations. L. delbrueckii and Lactococcus lactis bacteriophages were below the detection limit of the method used and high titres of cos type S. thermophilus bacteriophages were detected in only two cases. In one case a high titre of bacteriophages was concurrent with a S. thermophilus biotype shift in the culture

Multiple approaches to characterize the microbial community in a thermophilic anaerobic digester running on swine manure: a case study.

Science.gov (United States)

Tuan, Nguyen Ngoc; Chang, Yi-Chia; Yu, Chang-Ping; Huang, Shir-Ly

2014-01-01

In this study, the first survey of microbial community in thermophilic anaerobic digester using swine manure as sole feedstock was performed by multiple approaches including denaturing gradient gel electrophoresis (DGGE), clone library and pyrosequencing techniques. The integrated analysis of 21 DGGE bands, 126 clones and 8506 pyrosequencing read sequences revealed that Clostridia from the phylum Firmicutes account for the most dominant Bacteria. In addition, our analysis also identified additional taxa that were missed by the previous researches, including members of the bacterial phyla Synergistetes, Planctomycetes, Armatimonadetes, Chloroflexi and Nitrospira which might also play a role in thermophilic anaerobic digester. Most archaeal 16S rRNA sequences could be assigned to the order Methanobacteriales instead of Methanomicrobiales comparing to previous studies. In addition, this study reported that the member of Methanothermobacter genus was firstly found in thermophilic anaerobic digester. Copyright © 2014 Elsevier GmbH. All rights reserved.

Biosynthesis of cellulolytic enzymes by Aspergillus niger A. n. 33 and its selectants

Energy Technology Data Exchange (ETDEWEB)

Czajkowska, D.; Hornecka, D.; Ilnicka-Olejniczak, O.

1988-01-01

The aim of the investigations was to obtain - from the parent strain Aspergillus niger A.n. 33 - selectants with an increased ability of cellulolytic enzymes biosynthesis. Own selection methods allowed to receive two selectants A.n. 33/2 and A.n. 33/20 characterized by enhanced activities of saccharifying cellulase (respectively 0.11 and 0.14 FPU/cm/sup 3/), endo-beta-1,4-glucanase (15.4 and 21.8 U/cm/sup 3/) and cellobiase (0.6 and 1.4 IU/cm/sup 3/) as compared with the parent strain (FPA - 0.09 IU, CMC - 8.2 U and CB - 0.1 IU/cm/sup 3/). Moreover, the selectants differed in shape and size of conidial heads, in shape and colour of conidia, as well as in structure and shape of hyphase. Enzyme preparations obtained after ultrafiltration of liquid cultures were characterized by following activities: FPA-4-16 IU, CMC-900-1800 U, CB-60-120 IU and xylanase-250-280 IU/cm/sup 3/.

Effects of commercial pectolytic and cellulolytic enzyme preparations on the apple cell wall.

Science.gov (United States)

Dongowski, G; Sembries, S

2001-09-01

The action of three different commercial enzyme combinations on apple cell wall material has been examined in a model system under conditions of mash and pomace treatment by using an alcohol-insoluble substance prepared from apples. A part of the total dietary fiber, for example, galacturonan (pectin), appeared in the soluble fraction after enzymatic mash treatment. The soluble fraction increased intensely during pomace treatment. Furthermore, enzyme actions caused a change in the water-binding capacity of residues as well as changes in the monosaccharide composition and in the molecular weight distribution of saccharides in filtrates (soluble parts). The extent of decomposition of cell wall material and the increase of soluble oligomeric and/or polymeric dietary fiber components are caused by both the composition (pectinases, cellulases, and hemicellulases) and the activities of the enzyme preparations. The model experiments allow an insight into the reactions occurring during enzyme action on the plant cell wall, for example, during apple juice production using pectolytic and cellulolytic enzyme preparations.

Extremely thermophilic microorganisms and their polymer-hidrolytic enzymes

Directory of Open Access Journals (Sweden)

Andrade Carolina M.M.C.

1999-01-01

Full Text Available Thermophilic and hyperthermophilic microorganisms are found as normal inhabitants of continental and submarine volcanic areas, geothermally heated sea-sediments and hydrothermal vents and thus are considered extremophiles. Several present or potential applications of extremophilic enzymes are reviewed, especially polymer-hydrolysing enzymes, such as amylolytic and hemicellulolytic enzymes. The purpose of this review is to present the range of morphological and metabolic features among those microorganisms growing from 70oC to 100°C and to indicate potential opportunities for useful applications derived from these features.

Characterization of technetium(vII) reduction by cell suspensions of thermophilic bacteria and archaea.

Science.gov (United States)

Chernyh, Nikolay A; Gavrilov, Sergei N; Sorokin, Vladimir V; German, Konstantin E; Sergeant, Claire; Simonoff, Monique; Robb, Frank; Slobodkin, Alexander I

2007-08-01

Washed cell suspensions of the anaerobic hyperthermophilic archaea Thermococcus pacificus and Thermoproteus uzoniensis and the anaerobic thermophilic gram-positive bacteria Thermoterrabacterium ferrireducens and Tepidibacter thalassicus reduced technetium [(99)Tc(VII)], supplied as soluble pertechnetate with molecular hydrogen as an electron donor, forming highly insoluble Tc(IV)-containing grayish-black precipitate. Apart from molecular hydrogen, T. ferrireducens reduced Tc(VII) with lactate, glycerol, and yeast extract as electron donors, and T. thalassicus reduced it with peptone. Scanning electron microscopy and X-ray microanalysis of cell suspensions of T. ferrireducens showed the presence of Tc-containing particles attached to the surfaces of non-lysed cells. This is the first report on the reduction in Tc(VII) by thermophilic microorganisms of the domain Bacteria and by archaea of the phylum Euryarchaeota.

A robust nitrifying community in a bioreactor at 50 °C opens up the path for thermophilic nitrogen removal.

Science.gov (United States)

Courtens, Emilie Np; Spieck, Eva; Vilchez-Vargas, Ramiro; Bodé, Samuel; Boeckx, Pascal; Schouten, Stefan; Jauregui, Ruy; Pieper, Dietmar H; Vlaeminck, Siegfried E; Boon, Nico

2016-09-01

The increasing production of nitrogen-containing fertilizers is crucial to meet the global food demand, yet high losses of reactive nitrogen associated with the food production/consumption chain progressively deteriorate the natural environment. Currently, mesophilic nitrogen-removing microbes eliminate nitrogen from wastewaters. Although thermophilic nitrifiers have been separately enriched from natural environments, no bioreactors are described that couple these processes for the treatment of nitrogen in hot wastewaters. Samples from composting facilities were used as inoculum for the batch-wise enrichment of thermophilic nitrifiers (350 days). Subsequently, the enrichments were transferred to a bioreactor to obtain a stable, high-rate nitrifying process (560 days). The community contained up to 17% ammonia-oxidizing archaea (AOAs) closely related to 'Candidatus Nitrososphaera gargensis', and 25% nitrite-oxidizing bacteria (NOBs) related to Nitrospira calida. Incorporation of (13)C-derived bicarbonate into the respective characteristic membrane lipids during nitrification supported their activity as autotrophs. Specific activities up to 198±10 and 894±81 mg N g(-1) VSS per day for AOAs and NOBs were measured, where NOBs were 33% more sensitive to free ammonia. The NOBs were extremely sensitive to free nitrous acid, whereas the AOAs could only be inhibited by high nitrite concentrations, independent of the free nitrous acid concentration. The observed difference in product/substrate inhibition could facilitate the development of NOB inhibition strategies to achieve more cost-effective processes such as deammonification. This study describes the enrichment of autotrophic thermophilic nitrifiers from a nutrient-rich environment and the successful operation of a thermophilic nitrifying bioreactor for the first time, facilitating opportunities for thermophilic nitrogen removal biotechnology.

Continuous cultivation of a thermophilic bacterium Aeribacillus pallidus 418 for production of an exopolysaccharide applicable in cosmetic creams.

Science.gov (United States)

Radchenkova, N; Panchev, I; Vassilev, S; Kuncheva, M; Dobreva, S; Kambourova, M

2015-11-01

The aim of this study was to evaluate the effectiveness of continuous cultivation approach for exopolysaccharide (EPS) production by a thermophilic micro-organism and the potential of the synthesized EPS for application in cosmetic industry. Study on the ability of Aeribacillus pallidus 418, isolated as a good EPS producer, to synthesize the polymer in continuous cultures showed higher production in comparison with batch cultures. The degree of the EPS in the precipitate after continuous cultivation significantly increased. Non-Newtonian pseudoplastic and thixotropic behaviour of EPS determines the ability of the received cream to become more fluid after increasing time of application on the skin. This study demonstrates a highly efficient way for production of EPS from a continuous growth culture of A. pallidus 418 that have many advantages and can outperform batch culture by eliminating time for cleaning and sterilization of the vessel and the comparatively long lag phases before the organisms enter a brief period of high productivity. The valuable physico-chemical properties of the synthesized EPS influenced positively the properties of a commercial cream. EPSs from thermophilic micro-organisms are of special interest due to the advantages of the thermophilic processes and nonpathogenic nature of the polymer molecules. However, their industrial application is hindered by the comparatively low biomass and correspondingly EPS yield. Suggested continuous approach for EPS could have an enormous economic potential for an industrial scale production of thermophilic EPSs. © 2015 The Society for Applied Microbiology.

A Novel Process Configuration for Anaerobic Digestion of Source-Sorted Household Waste Using Hyper-Thermophilic Post-Treatment

DEFF Research Database (Denmark)

Hartmann, H.; Ahring, Birgitte Kiær

2005-01-01

A novel reactor configuration was investigated for anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW). An anaerobic hyper-thermophilic (68°C) reactor R68 was implemented as a post–treatment step for the effluent of a thermophilic reactor R1 (55°C) in order to enhance...... hydrolysis of recalcitrant organic matter, improve sanitation and ease the stripping of ammonia from the reactor. The efficiency of the combined system was studied in terms of methane yield, volatile solids (VS) reduction and volatile fatty acid (VFA) production at different hydraulic retention times (HRT...

Thermophilic fermentation of acetoin and 2,3-butanediol by a novel Geobacillus strain

Directory of Open Access Journals (Sweden)

Xiao Zijun

2012-12-01

Full Text Available Abstract Background Acetoin and 2,3-butanediol are two important biorefinery platform chemicals. They are currently fermented below 40°C using mesophilic strains, but the processes often suffer from bacterial contamination. Results This work reports the isolation and identification of a novel aerobic Geobacillus strain XT15 capable of producing both of these chemicals under elevated temperatures, thus reducing the risk of bacterial contamination. The optimum growth temperature was found to be between 45 and 55°C and the medium initial pH to be 8.0. In addition to glucose, galactose, mannitol, arabionose, and xylose were all acceptable substrates, enabling the potential use of cellulosic biomass as the feedstock. XT15 preferred organic nitrogen sources including corn steep liquor powder, a cheap by-product from corn wet-milling. At 55°C, 7.7 g/L of acetoin and 14.5 g/L of 2,3-butanediol could be obtained using corn steep liquor powder as a nitrogen source. Thirteen volatile products from the cultivation broth of XT15 were identified by gas chromatography–mass spectrometry. Acetoin, 2,3-butanediol, and their derivatives including a novel metabolite 2,3-dihydroxy-3-methylheptan-4-one, accounted for a total of about 96% of all the volatile products. In contrast, organic acids and other products were minor by-products. α-Acetolactate decarboxylase and acetoin:2,6-dichlorophenolindophenol oxidoreductase in XT15, the two key enzymes in acetoin metabolic pathway, were found to be both moderately thermophilic with the identical optimum temperature of 45°C. Conclusions Geobacillus sp. XT15 is the first naturally occurring thermophile excreting acetoin and/or 2,3-butanediol. This work has demonstrated the attractive prospect of developing it as an industrial strain in the thermophilic fermentation of acetoin and 2,3-butanediol with improved anti-contamination performance. The novel metabolites and enzymes identified in XT15 also indicated its

Dietary Corn Bran Fermented by Bacillus subtilis MA139 Decreased Gut Cellulolytic Bacteria and Microbiota Diversity in Finishing Pigs

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Ping Liu

2017-12-01

Full Text Available Solid-state fermentation of feedstuffs by Bacillus subtilis MA139 can reduce insoluble dietary fiber content in vitro and improve growth performance in pigs. This study was conducted to investigate the effects of dietary corn bran (CB fermented by B. subtilis on growth performance and gut microbiota composition in finishing pigs. A total of 60 finishing pigs were allocated to 3 dietary treatments consisting of a control (CON diet, a 10% CB diet, and a 10% fermented CB (FCB diet in a 21 d feeding trial. Growth performance and nutrient digestibility were evaluated. Fecal samples were determined for bacterial community diversity by 16S rRNA gene amplicon sequencing. The dietary CB and FCB did not affect growth performance of finishing pigs. The digestibility of organic matter was decreased in both CB and FCB treatments compared with CON group (P < 0.05. The α-diversity for bacterial community analysis of Chao 1 in FCB treatment was lower than CON treatment (P < 0.05. The Fibrobacteres phylum belongs to cellulolytic bacteria was isolated, and their relative abundance in CB group showed no difference between CON and FCB treatments. The abundance of Lachnospiraceae_NK4A136_group in CB treatment was higher than CON and FCB groups (P < 0.05, whereas the population of norank_f_Prevotellaceae was higher in FCB group compared to CON and CB groups (P < 0.05. In conclusion, dietary FCB decreased the abundance of bacterial communities, particularly the population of bacteria related to cellulolytic degradation.

A strict anaerobic extreme thermophilic hydrogen-producing culture enriched from digested household waste

DEFF Research Database (Denmark)

Karakashev, Dimitar Borisov; Kotay, Shireen Meher; Trably, Eric

2009-01-01

The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H-2) producers from digested household solid wastes. A strict anaerobic extreme thermophilic H-2 producing bacterial culture was enriched from a lab-scale digester treating household...... wastes at 70 degrees C. The enriched mixed culture consisted of two rod-shaped bacterial members growing at an optimal temperature of 80 degrees C and an optimal pH 8.1. The culture was able to utilize glucose, galactose, mannose, xylose, arabinose, maltose, sucrose, pyruvate and glycerol as carbon...... sources. Growth on glucose produced acetate, H-2 and carbon dioxide. Maximal H-2 production rate on glucose was 1.1 mmol l(-1) h(-1) with a maximum H-2 yield of 1.9 mole H-2 per mole glucose. 16S ribosomal DNA clone library analyses showed that the culture members were phylogenetically affiliated...

Effect of alkaline pretreatment on mesophilic and thermophilic anaerobic digestion of a submerged macrophyte: Inhibition and recovery against dissolved lignin during semi-continuous operation.

Science.gov (United States)

Koyama, Mitsuhiko; Watanabe, Keiko; Kurosawa, Norio; Ishikawa, Kanako; Ban, Syuhei; Toda, Tatsuki

2017-08-01

The long-term effect of alkaline pretreatment on semi-continuous anaerobic digestion (AD) of the lignin-rich submerged macrophyte Potamogeton maackianus was investigated using mesophilic and thermophilic conditions. In pretreated reactors, dissolved lignin accumulated to high levels. CH 4 production under the pretreated condition was higher than that of the untreated condition, but decreased from Days 22 (mesophilic) and 42 (thermophilic). However, CH 4 production subsequently recovered, although dissolved lignin accumulated. Further, the change in the microbial community was observed between conditions. These results suggest that dissolved lignin temporarily inhibited AD, although acclimatization to dissolved lignin occurred during long-term operation. During the steady state period, mesophilic conditions achieved a 42% increase in the CH 4 yield using pretreatment, while thermophilic conditions yielded an 8% increment. Because volatile fatty acids accumulated even after acclimatization during the thermophilic pretreated condition and was discharged with the effluent, improvement of the methanogenic step would enable enhanced CH 4 recovery. Copyright © 2017 Elsevier Ltd. All rights reserved.

Filamentous Growth in Eremothecium Fungi

DEFF Research Database (Denmark)

Oskarsson, Therese

, this thesis deals with some of the aspects of hyphal growth, which is an important virulence factor for pathogenic fungi infecting both humans and plants. Hyphal establishment through continuous polar growth is a complex process, requiring the careful coordination of a large subset of proteins involved......-regulatory activity of AgGts1, the protein could have additional actin organizing properties. In the second and third part, this thesis addresses the use of A. gossypii and its relative E. cymbalariae as model organisms for filamentous growth. A series of assays analyzed the capability of Eremothecium genus fungi...... of molecular tools for E. cymbalariae to enable a faster and more efficient approach for genetic comparisons between Eremothecium genus fungi....

The diversity and specificity of the extracellular proteome in the cellulolytic bacterium Caldicellulosiruptor bescii is driven by the nature of the cellulosic growth substrate.

Science.gov (United States)

Poudel, Suresh; Giannone, Richard J; Basen, Mirko; Nookaew, Intawat; Poole, Farris L; Kelly, Robert M; Adams, Michael W W; Hettich, Robert L

2018-01-01

Caldicellulosiruptor bescii is a thermophilic cellulolytic bacterium that efficiently deconstructs lignocellulosic biomass into sugars, which subsequently can be fermented into alcohols, such as ethanol, and other products. Deconstruction of complex substrates by C. bescii involves a myriad of highly abundant, substrate-specific extracellular solute binding proteins (ESBPs) and carbohydrate-active enzymes (CAZymes) containing carbohydrate-binding modules (CBMs). Mass spectrometry-based proteomics was employed to investigate how these substrate recognition proteins and enzymes vary as a function of lignocellulosic substrates. Proteomic analysis revealed several key extracellular proteins that respond specifically to either C5 or C6 mono- and polysaccharides. These include proteins of unknown functions (PUFs), ESBPs, and CAZymes. ESBPs that were previously shown to interact more efficiently with hemicellulose and pectin were detected in high abundance during growth on complex C5 substrates, such as switchgrass and xylan. Some proteins, such as Athe_0614 and Athe_2368, whose functions are not well defined were predicted to be involved in xylan utilization and ABC transport and were significantly more abundant in complex and C5 substrates, respectively. The proteins encoded by the entire glucan degradation locus (GDL; Athe_1857, 1859, 1860, 1865, 1867, and 1866) were highly abundant under all growth conditions, particularly when C. bescii was grown on cellobiose, switchgrass, or xylan. In contrast, the glycoside hydrolases Athe_0609 (Pullulanase) and 0610, which both possess CBM20 and a starch binding domain, appear preferential to C5/complex substrate deconstruction. Some PUFs, such as Athe_2463 and 2464, were detected as highly abundant when grown on C5 substrates (xylan and xylose), also suggesting C5-substrate specificity. This study reveals the protein membership of the C. bescii secretome and demonstrates its plasticity based on the complexity (mono

Temporal change of composition and potential activity of the thermophilic archaeal community during the composting of organic material.

Science.gov (United States)

Thummes, Kathrin; Kämpfer, Peter; Jäckel, Udo

2007-07-01

To date, composting has been regarded as an aerobic process but it has been shown that composting piles are often sources of atmospheric methane. In order to gain a more comprehensive view on the diversity of methanogenic Archaea in compost, gas chromatographical methods and molecular cloning were used to study relationships of thermophilic archaeal communities and changes in methane production potential during compost maturation. According to the thermophilic methane production potential, wide differences could be detected between differently aged compost materials. In material derived from 3- and 4-week-old piles, low and no thermophilic methane production potential, respectively, was observed at 50 degrees C. Material from a 6-week-old pile showed the maximum methane production. With compost maturation, the production slowly decreased again with 6 weeks, 8 weeks, and mature compost showing an optimum methane production potential at 60 degrees C. At 70 degrees C, only 6-week-old material showed a comparable high production of methane. The 16S rRNA-based phylogenetic surveys revealed an increase of archaeal diversity with compost maturation. In the 6-week-old material, 86% of the sequences in the archaeal 16S rRNA library had the highest sequence similarities to Methanothermobacter spp. and the remaining 14% of the clones were related to Methanosarcina thermophila. Quantification of methanogens in 6-week-old material, on the basis of the methane production rate, resulted in values of about 2x10(7) cells per gram fresh weight. In 8-week-old and mature compost material, the proportion of sequences similar to Methanothermobacter spp. decreased to 34% and 0%, respectively. The mature compost material showed the highest variation in identified sequences, although 33% could be assigned to as yet uncultured Archaea (e.g. Rice cluster I, III, and IV). Our results indicate that compost harbours a diverse community of thermophilic methanogens, with changing composition

Cellulolytic activities of wild type fungi isolated from decayed wood ...

African Journals Online (AJOL)

The mycological profile of decayed wood cuttings sourced from a saw mill located at Uwasota Road, Benin City was investigated using serial dilution and pour plate techniques. The mean fungal counts ranged from 0.9 ×106 cfu/g to 2.7 ×106 cfu/g respectively. Four fungal species were identified; Aspergillus niger, ...

Heteroresistance and fungi.

Science.gov (United States)

Ferreira, Gabriella F; Santos, Daniel A

2017-09-01

The concept of heteroresistance refers to the heterogeneous susceptibility to an antimicrobial drug in a microorganism population, meaning that some clones may be resistant and others are susceptible. This phenomenon has been widely studied in bacteria, but little attention has been given to its expression in fungi. We review the available literature on heteroresistance in fungi and invite the reader to recognise this phenomenon as a fungal mechanism to adapt to environmental stress, which may interfere both in resistance and virulence. Finally, heteroresistance may explain the treatment failures to eradicate mycosis in some patients treated with a seemingly appropriate antifungal. © 2017 Blackwell Verlag GmbH.

Use of Aspergillus niger for bioconversion of apple distillery waste

Energy Technology Data Exchange (ETDEWEB)

Friedrich, J.; Cimerman, A.; Perdih, A.

1983-01-01

The bioconversion of waste material remaining after apple brandy distillation was investigated. Different cellulolytic fungi were tested for their ability to convert the waste organic substances into microbial biomass. An Aspergillus niger strain was chosen as the most convenient microorganism. By growing this mold on the apple slop the following results were obtained: filtration time was shortened by 30 times, reduction of the chemical oxygen demand in the liquid phase in the range of 50-80% depending on the substrate dilution and a dry filter cake enriched with fungal biomass to about 12 g/l containing up to 22% raw proteins and certain amounts of cellulolytic enzymes in the filtrate. The influence of the initial pH, the salt addition and the dilution of the substrate were studied as well. 12 references.

Cellulose digestion in Monochamus marmorator Kby. (coleoptera: Cerambycidae): role of acquired fungal enzymes

Energy Technology Data Exchange (ETDEWEB)

Kukol, J.J.; Martin, M.M.

1986-05-01

Larvae of the balsam fir sawyer, Monochamus marmorator Kby. (Coleoptera, Cerambycidae), contain midgut digestive enzymes active against hemicellulose and cellulose. Cellulases from larvae fed on balsam fir wood infected with the fungus, Trichoderma harzianum Rifai (Deuteromycetes, Moniliales, Moniliaceae), were found to be identical to those of the cellulase complex produced by this fungus when compared using chromatography, electrophoresis, and isofocusing. When larvae are maintained on a fungusfree diet, their midgut fluids lack cellulolytic activity, and they are unable to digest cellulose. Cellulolytic capacity can be restored by feeding the larvae wood permeated by fungi. We conclude that the enzymes which enable M. marmorator larvae to digest cellulose are not produced by the larvae. Instead, the larvae acquire the capacity to digest cellulose by ingesting active fungal cellulases while feeding in fungus-infected wood.

Cellulose digestion in Monochamus marmorator Kby. (coleoptera: Cerambycidae): role of acquired fungal enzymes

International Nuclear Information System (INIS)

Kukol, J.J.; Martin, M.M.

1986-01-01

Larvae of the balsam fir sawyer, Monochamus marmorator Kby. (Coleoptera, Cerambycidae), contain midgut digestive enzymes active against hemicellulose and cellulose. Cellulases from larvae fed on balsam fir wood infected with the fungus, Trichoderma harzianum Rifai (Deuteromycetes, Moniliales, Moniliaceae), were found to be identical to those of the cellulase complex produced by this fungus when compared using chromatography, electrophoresis, and isofocusing. When larvae are maintained on a fungusfree diet, their midgut fluids lack cellulolytic activity, and they are unable to digest cellulose. Cellulolytic capacity can be restored by feeding the larvae wood permeated by fungi. We conclude that the enzymes which enable M. marmorator larvae to digest cellulose are not produced by the larvae. Instead, the larvae acquire the capacity to digest cellulose by ingesting active fungal cellulases while feeding in fungus-infected wood

Effects of Atrazine on Soil Microorganisms

Directory of Open Access Journals (Sweden)

Ljiljana Radivojević

2006-01-01

Full Text Available Effects of the herbicide atrazine on soil microorganisms was investigated. Trials were set up in laboratory, on a clay loam soil. Atrazine was applied at 8.0, 40.0 and 80.0 mg/kg soil rates. The abundance of total microorganisms, fungi, actinomycetes, cellulolytic microorganisms and amino-heterotrophs was recorded. Soil samples were collected 1, 7, 14, 21, 30 and 60 days after atrazine treatment for microbiological analyses.The results showed that the intensity of atrazine effect on soil microorganisms depended on treatment rate, exposure time and group of microorganisms. Atrazine had an inhibiting effect on cellulolytic microorganisms and amino-heterotrophs. Initially, it inhibited fungiand actinomycetes but its effect turned into a stimulating one once a population recovered. Atrazine had a stimulating effect on total abundance of microorganisms.

Effect of feed to microbe ratios on anaerobic digestion of Chinese cabbage waste under mesophilic and thermophilic conditions: biogas potential and kinetic study.

Science.gov (United States)

Kafle, Gopi Krishna; Bhattarai, Sujala; Kim, Sang Hun; Chen, Lide

2014-01-15

The objective of this study was to investigate the effect of the feed-to-microbe (F/M) ratios on anaerobic digestion of Chinese cabbage waste (CCW) generated from a kimchi factory. The batch test was conducted for 96 days under mesophilic (36.5 °C) (Experiment I) and thermophilic (55 °C) conditions (Experiment II) at F/M ratios of 0.5, 1.0 and 2.0. The first-order kinetic model was evaluated for methane yield. The biogas yield in terms of volatile solids (VS) added increased from 591 to 677 mL/g VS under mesophilic conditions and 434 to 639 mL/g VS under thermophilic conditions when the F/M ratio increased from 0.5 to 2.0. Similarly, the volumetric biogas production increased from 1.479 to 6.771 L/L under mesophilic conditions and from 1.086 to 6.384 L/L under thermophilic conditions when F/M ratio increased from 0.5 to 2.0. The VS removal increased from 59.4 to 75.6% under mesophilic conditions and from 63.5 to 78.3% under thermophilic conditions when the F/M ratio increased from 0.5 to 2.0. The first-order kinetic constant (k, 1/day) decreased under the mesophilic temperature conditions and increased under thermophilic conditions when the F/M ratio increased from 0.5 to 2.0. The difference between the experimental and predicted methane yield was in the range of 3.4-14.5% under mesophilic conditions and in the range of 1.1-3.0% under thermophilic conditions. The predicted methane yield derived from the first-order kinetic model was in good agreement with the experimental results. Published by Elsevier Ltd.

Sequential Detection of Thermophilic Lipase and Protease by Zymography.

Science.gov (United States)

Kurz, Liliana; Hernández, Zully; Contreras, Lellys M; Wilkesman, Jeff

2017-01-01

Lipase and protease present in cell-free fractions of thermophilic Bacillus sp. cultures were analyzed by polyacrylamide gel (PAG) electrophoresis. After run, the gel is electrotransferred to another PAG copolymerized with glycerol tributyrate, olive oil, and gelatin. This multi-substrate gel was incubated first for lipase detection, until bands appeared, and then stained with Coomassie for protease detection. Advantages of this sequential procedure are the detection of two different enzyme activities on a single PAG, beside time and resource saving.

Acquired Thermotolerance and Heat Shock Proteins in Thermophiles from the Three Phylogenetic Domains

DEFF Research Database (Denmark)

Trent, Jonathan D.; Gabrielsen, Mette; Jensen, Bo

1994-01-01

Thermophilic organisms from each of the three phylogenetic domains (Bacteria, Archaea, and Eucarya) acquired thermotolerance after heat shock. Bacillus caldolyticus grown at 60 degrees C and heat shocked at 69 degrees C for 10 min showed thermotolerance at 74 degrees C, Sulfolobus shibatae grown...

Hydrogen limitation and syntrophic growth among natural assemblages of thermophilic methanogens at deep-sea hydrothermal vents

Directory of Open Access Journals (Sweden)

Begüm D. Topçuoğlu

2016-08-01

Full Text Available Thermophilic methanogens are common autotrophs at hydrothermal vents, but their growth constraints and dependence on H2 syntrophy in situ are poorly understood. Between 2012 and 2015, methanogens and H2-producing heterotrophs were detected by growth at 80°C and 55°C at most diffuse (7-40°C hydrothermal vent sites at Axial Seamount. Microcosm incubations of diffuse hydrothermal fluids at 80°C and 55°C demonstrated that growth of thermophilic and hyperthermophilic methanogens is primarily limited by H2 availability. Amendment of microcosms with NH4+ generally had no effect on CH4 production. However, annual variations in abundance and CH4 production were observed in relation to the eruption cycle of the seamount. Microcosm incubations of hydrothermal fluids at 80°C and 55°C supplemented with tryptone and no added H2 showed CH4 production indicating the capacity in situ for methanogenic H2 syntrophy. 16S rRNA genes were found in 80°C microcosms from H2-producing archaea and H2-consuming methanogens, but not for any bacteria. In 55°C microcosms, sequences were found from the H2-producing bacteria and H2-consuming methanogens and sulfate-reducing bacteria. A co-culture of representative organisms showed that Thermococcus paralvinellae supported the syntrophic growth of Methanocaldococcus bathoardescens at 82°C and Methanothermococcus sp. strain BW11 at 60°C. The results demonstrate that modeling of subseafloor methanogenesis should focus primarily on H2 availability and temperature, and that thermophilic H2 syntrophy can support methanogenesis within natural microbial assemblages and may be an important energy source for thermophilic autotrophs in marine geothermal environments.

Influence of variable feeding on mesophilic and thermophilic co-digestion of Laminaria digitata and cattle manure

International Nuclear Information System (INIS)

Sarker, Shiplu; Møller, Henrik Bjarne; Bruhn, Annette

2014-01-01

Highlights: • Anaerobic co-digestion of L. digitata and cattle manure, at ∼35 and ∼50 °C. • Mesophilic co-digestion showed somewhat stable specific methane, but increased volumetric yield. • Thermophilic co-digester yielded higher methane at higher input of algae compared to control. • Mesophilic co-digester performed better in terms of various parameters except methane yield. - Abstract: In this study the effect of various feeding ratios on mesophilic (∼35 °C) and thermophilic (∼50 °C) co-digestion of brown algae Laminaria digitata and cattle manure was investigated. Algae input of 15% VS caused no influence on specific methane yield from mesophilic co-digester while deteriorated the process parameters such as the development of propionic acid in total volatile fatty acids (tVFA) pattern of the thermophilic co-digester. The accumulation of tVFA continued for the latter reactor as the feeding ratio of algae enhanced to 24% VS, but the specific methane yield improved dramatically. Same rise in feeding once again showed no improvement in specific methane yield from mesophilic co-digester even though the other process parameters stabilized or, enriched such as the gain in average volumetric methane yield. For the last feeding ratio at 41% VS algae, specific methane yield from mesophilic co-digester slightly increased which however was not still comparable with the ultimate methane yield from the cattle manure alone. The thermophilic co-digestion on the other hand yielded maximum specific methane, together with the improvement in different process characteristics, as the feeding of algae maximized at the final stage. The trend of methane production from this reactor nevertheless was sharply downward towards the end of the experiment suggesting that the optimum feeding ratio has already been achieved for the present experimental conditions

Biochemical characterization of a thermophilic β-mannanase from Talaromyces leycettanus JCM12802 with high specific activity.

Science.gov (United States)

Wang, Caihong; Luo, Huiying; Niu, Canfang; Shi, Pengjun; Huang, Huoqing; Meng, Kun; Bai, Yingguo; Wang, Kun; Hua, Huifang; Yao, Bin

2015-02-01

Thermophilic β-mannanases are of increasing importance for wide industrial applications. In the current study, gene cloning, functional expression in Pichia pastoris, and characterization of a thermophilic β-mannanase (Man5A) from thermophilic Talaromyces leycettanus JCM12802 are reported. Deduced Man5A exhibits the highest identity with a putative β-mannanase from Talaromyces stipitatus ATCC10500 (70.3 %) and is composed of an N-terminal signal peptide, a fungal-type carbohydrate-binding module (CBM) of family 1, and a catalytic domain of glycosyl hydrolase (GH) family 5 at the C-terminus. Two recombinant proteins with different glycosylation levels, termed Man5A1 (72 kDa) and Man5A2 (60 kDa), were identified after purification. Both enzymes were thermophilic, exhibiting optimal activity at 85-90 °C, and were highly stable at 70 °C. Man5A1 and Man5A2 had a pH optimum of 4.5 and 4.0, respectively, and were highly stable over the broad pH range of 3.0-10.0. Most metal ions and sodium dodecyl sulfate (SDS) had no effect on the enzymatic activities. Man5A1 and Man5A2 exhibited high specific activity (2,160 and 1,800 U/mg, respectively) when using locust bean gum as the substrate. The CBM1 and two key residues D191 and R286 were found to affect Man5A thermostability. Man5A displays a classical four-site-binding mode, hydrolyzing mannooligosaccharides into smaller units, galactomannan into mannose and mannobiose, and glucomanman into mannose, mannobiose, and mannopentaose, respectively. All these properties make Man5A a good candidate for extensive applications in the bioconversion, pulp bleaching, textile, food, and feed industries.

Differential antibiotic sensitivity determined by the large ribosomal subunit in thermophilic archaea.

OpenAIRE

Ruggero, D; Londei, P

1996-01-01

Hybrid ribosomes obtained by mixing the ribosomal subunits of the extremely thermophilic archaea Sulfolobus solfataricus and Desulfurococcus mobilis were tested for their sensitivity to selected antibiotics. It is shown that structural differences in the large ribosomal subunits determine qualitatively and quantitatively the patterns of response to alpha-sarcin and paromomycin in these species.

Production and assay of cellulolytic enzyme activity of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya abbatoir, Indonesia

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W. P. Lokapirnasari

2015-03-01

Full Text Available Aim: This study aims to produce and assay cellulolytic enzyme activity (endo-(1,4-β-D-glucanase, exo-(1,4-β-Dglucanase, and β-glucosidase, at optimum temperature and optimum pH of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya Abbatoir, Indonesia. Materials and Methods: To produce enzyme from a single colony of E. cloacae WPL 214, 98 × 1010 CFU/ml of isolates was put into 20 ml of liquid medium and incubated in a shaker incubator for 16 h at 35°C in accordance with growth time and optimum temperature of E. cloacae WPL 214. Further on, culture was centrifuged at 6000 rpm at 4°C for 15 min. Pellet was discarded while supernatant containing cellulose enzyme activity was withdrawn to assay endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase, and β-glucosidase. Results: Cellulase enzyme of E. cloacae WPL 214 isolates had endoglucanase activity of 0.09 U/ml, exoglucanase of 0.13 U/ml, and cellobiase of 0.10 U/ml at optimum temperature 35°C and optimum pH 5. Conclusion: E. cloacae WPL 214 isolated from bovine rumen fluid waste produced cellulose enzyme with activity as cellulolytic enzyme of endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase and β-glucosidase.

Pyrophosphate as a central energy carrier in the hydrogen-producing extremely thermophilic Caldicellulosiruptor saccharolyticus

NARCIS (Netherlands)

Bielen, A.A.M.; Willquist, K.; Engman, J.; Oost, van der J.; Niel, van E.W.J.; Kengen, S.W.M.

2010-01-01

The role of inorganic pyrophosphate (PPi) as an energy carrier in the central metabolism of the extremely thermophilic bacterium Caldicellulosiruptor saccharolyticus was investigated. In agreement with its annotated genome sequence, cell extracts were shown to exhibit PPi-dependent

Hydrogen production from carrot pulp by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

NARCIS (Netherlands)

Vrije, de G.J.; Budde, M.A.W.; Lips, S.J.J.; Bakker, R.R.; Mars, A.E.; Claassen, P.A.M.

2010-01-01

Hydrogen was produced from carrot pulp hydrolysate, untreated carrot pulp and (mixtures of) glucose and fructose by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana in pH-controlled bioreactors. Carrot pulp hydrolysate was obtained after enzymatic hydrolysis

[Conversion of acetic acid to methane by thermophiles]. Progress report, May 15, 1989--May 14, 1993

Energy Technology Data Exchange (ETDEWEB)

Zinder, S.H.

1993-06-01

The primary goal of this project is to obtain a better understanding of thermophilic microorganisms which convert acetic acid to CH{sub 4}. The previous funding period represents a departure from earlier research in this laboratory, which was more physiological and ecological. The present work is centered on the biochemistry of the thermophile Methanothrix sp. strain CALS-1. this organism presents a unique opportunity, with its purity and relatively rapid growth, to do comparative biochemical studies with the other major acetotrophic genus Methanosarcina. We previously found that Methanothrix is capable of using acetate at concentrations 100 fold lower than Methanosarcina. This finding suggests that there are significant differences in the pathways of methanogenesis from acetate in the two genera.

Mesophilic and thermophilic anaerobic co-digestion of winery wastewater sludge and wine lees: An integrated approach for sustainable wine production.

Science.gov (United States)

Da Ros, C; Cavinato, C; Pavan, P; Bolzonella, D

2017-12-01

In this work, winery wastes generated by a cellar producing approximately 300,000 hL of wine per year was monitored for a period of one year. On average, 196 L of wastewater, 0.1 kg of waste activated sludge (dry matter) and 1.6 kg of wine lees were produced per hectoliter of wine produced. Different winery wastes, deriving from different production steps, namely waste activated sludge from wastewater treatment and wine lees, were co-treated using an anaerobic digestion process. Testing was conducted on a pilot scale for both mesophilic and thermophilic conditions. The process was stable for a long period at 37 °C, with an average biogas production of 0.386 m 3 /kg COD fed . On the other hand, for thermophilic conditions, volatile fatty acids accumulated in the reactor and the process failed after one hydraulic retention time (23 days). In order to fix the biological process, trace elements (iron, cobalt and nickel) were added to the feed of the thermophilic reactor. Metals augmentation improved process stability and yields at 55 °C. The pH ranged between 7.8 and 8.0, and specific gas production was 0.450 m 3 /kg COD fed , which corresponded to dry matter and COD removals of 34% and 88%, respectively. Although the observed performances in terms of biogas production were good, the thermophilic process exhibited some limitations related to both the necessity of metals addition and the worse dewaterability properties. In fact, while the mesophilic digestates reached a good dewatering quality via the addition of 6.5 g of polymer per kg of dry matter, the required dosage for the thermophilic sludge was greater than 10 g/kg of dry matter. Copyright © 2016 Elsevier Ltd. All rights reserved.

Studies on certain aspects of seed-borne fungi. VI. Fungi associated with different cultivars of wheat (Triticum aestivum L.)

OpenAIRE

K. K. Pandey

2014-01-01

Fungi associated with eight cultivars of wheat have been investigated. Twenty seven species were isolated from external and internal surface of all the wheat (Triticum aestivum L.) cultivars respectively. Out of five dominant and subdominant fungi anly Aspergillus terreus and Alternaria tenuis were able to colonize internally. The culture filtrates of test fungi reduced the germination of all wheat varieties up to different degrees.

Microbial degradation of coconut coir dust for biomass production

Energy Technology Data Exchange (ETDEWEB)

Uyenco, F.R.; Ochoa, J.A.K.

Several species of white-rot fungi were studied for its ability to degrade the lignocellulose components of coir dust at optimum conditions. The most effective fungi was Phanerochaeta chrysosporium UPCC 4003. This organism degraded the lignocellulose complex of coir dust at a rate of about 25 percent in 4 weeks. The degradation process was carried on with minimal nitrogen concentration, coconut water supplementation and moisture levels between 85-90 percent. Shake flask cultures of the degraded coir dust using cellulolytic fungi were not effective. In fermentor cultures with Chaetomium cellulolyticum UPCC 3934, supplemented coir dust was converted into a microbial biomass product (MBP) with 15.58 percent lignin, 19.20 percent cellulose and 18.87 percent protein. More work is being done on the utilization of coir dust on a low technology.

Effects of seed sludge on fermentative characteristics and microbial community structures in thermophilic hydrogen fermentation of starch

Energy Technology Data Exchange (ETDEWEB)

Akutsu, Yohei; Tandukar, Madan; Kubota, Kengo; Harada, Hideki [Department of Civil and Environmental Engineering, Tohoku University, 6-6-06 Aoba, Sendai, Miyagi 980-8579 (Japan); Li, Yu-You [Department of Civil and Environmental Engineering, Tohoku University, 6-6-06 Aoba, Sendai, Miyagi 980-8579 (Japan); Department of Urban and Environmental Engineering, Tianjin Institute of Urban Construction, Jinjinggonglu 26, Tianjin, 300384 (China)

2008-11-15

In this work, effects of seed sludge on thermophilic hydrogen fermentation were investigated. Five different kinds of sludge were separately inoculated in completely stirred tank reactors (CSTRs), without any pretreatment. The reactors were operated in parallel with starch as substrate under thermophilic condition (55 C) at a hydraulic retention time (HRT) of 24 h for 50 days. Stable hydrogen production was achieved in all reactors. The highest hydrogen yield (2.32 mol H{sub 2}/mol glucose) was obtained from the reactor seeded with thermophilically digested activated sludge. The stoichiometric equation of hydrogen fermentation calculated using the data obtained in this study was expressed as follows: starch ([C{sub 6}H{sub 10}O{sub 5}]n) {yields} 2.32n H{sub 2} + 2.14n CO{sub 2} + 0.50n acetate + 0.63n butyrate + 0.11n cell (C{sub 5}H{sub 7}NO{sub 2}). Microbial community structure was analyzed by polymerase chain reaction-denaturing gel gradient electrophoresis (PCR-DGGE) and 16S rRNA gene cloning. Close relatives of the Thermoanaerobacterium were found to be the most predominated one in all reactors. (author)

Seasonal variability of thermophilic Campylobacter spp. in raw milk sold by automatic vending machines in Lombardy Region

Directory of Open Access Journals (Sweden)

Barbara Bertasi

2016-06-01

Full Text Available In temperate climates, a seasonal trend was observed in the incidence of human campylobacteriosis cases, with peaks reported in spring and autumn in some countries, or in summer in others; a similar trend was observed in Campylobacter spp. dairy cattle faecal shedding, suggesting that cattle may play a role in the seasonal peak of human infection. The objectives of this study were to assess if a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk exists and to evaluate a possible relation between this and the increase of human campylobacteriosis incidence in summer months. The results showed a mean prevalence of 1.6% of milk samples positive for thermophilic Campylobacter spp. with a wide range (0.0-3.1% in different months during the three years considered. The statistical analysis showed a significant difference (PCampylobacter spp. between warmer and cooler months (2.3 vs 0.6%. The evidence of a seasonal trend in thermophilic Campylobacter spp. contamination of raw milk sold for direct consumption, with an increase of the prevalence in warmer months, may represent one of the possible links between seasonal trend in cattle faecal shedding and seasonal trend in human campylobacteriosis.

Potential Antiviral Agents from Marine Fungi: An Overview

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Soheil Zorofchian Moghadamtousi

2015-07-01

Full Text Available Biodiversity of the marine world is only partially subjected to detailed scientific scrutiny in comparison to terrestrial life. Life in the marine world depends heavily on marine fungi scavenging the oceans of lifeless plants and animals and entering them into the nutrient cycle by. Approximately 150 to 200 new compounds, including alkaloids, sesquiterpenes, polyketides, and aromatic compounds, are identified from marine fungi annually. In recent years, numerous investigations demonstrated the tremendous potential of marine fungi as a promising source to develop new antivirals against different important viruses, including herpes simplex viruses, the human immunodeficiency virus, and the influenza virus. Various genera of marine fungi such as Aspergillus, Penicillium, Cladosporium, and Fusarium were subjected to compound isolation and antiviral studies, which led to an illustration of the strong antiviral activity of a variety of marine fungi-derived compounds. The present review strives to summarize all available knowledge on active compounds isolated from marine fungi with antiviral activity.

Marine Fungi: A Source of Potential Anticancer Compounds

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Sunil K. Deshmukh

2018-01-01

Full Text Available Metabolites from marine fungi have hogged the limelight in drug discovery because of their promise as therapeutic agents. A number of metabolites related to marine fungi have been discovered from various sources which are known to possess a range of activities as antibacterial, antiviral and anticancer agents. Although, over a thousand marine fungi based metabolites have already been reported, none of them have reached the market yet which could partly be related to non-comprehensive screening approaches and lack of sustained lead optimization. The origin of these marine fungal metabolites is varied as their habitats have been reported from various sources such as sponge, algae, mangrove derived fungi, and fungi from bottom sediments. The importance of these natural compounds is based on their cytotoxicity and related activities that emanate from the diversity in their chemical structures and functional groups present on them. This review covers the majority of anticancer compounds isolated from marine fungi during 2012–2016 against specific cancer cell lines.

Feasibility of thermophilic anaerobic processes for treating waste activated sludge under low HRT and intermittent mixing.

Science.gov (United States)

Leite, Wanderli; Magnus, Bruna Scandolara; Guimarães, Lorena Bittencourt; Gottardo, Marco; Belli Filho, Paulo

2017-10-01

Thermophilic anaerobic digestion (AD) arises as an optimized solution for the waste activated sludge (WAS) management. However, there are few feasibility studies using low solids content typically found in the WAS, and that consider uncommon operational conditions such as intermittent mixing and low hydraulic retention time (HRT). In this investigation, a single-stage pilot reactor was used to treat WAS at low HRT (13, 9, 6 and 5 days) and intermittent mixing (withholding mixing 2 h prior feeding). Thermophilic anaerobic digestion (55 °C) was initiated from a mesophilic digester (35 °C) by the one-step startup strategy. Although instabilities on partial alkalinity (1245-3000 mgCaCO 3 /L), volatile fatty acids (1774-6421 mg/L acetic acid) and biogas production (0.21-0.09 m 3 /m 3 reactor .d) were observed, methanogenesis started to recover in 18 days. The thermophilic treatment of WAS at 13 and 9 days HRT efficiently converted VS into biogas (22 and 21%, respectively) and achieved high biogas yield (0.24 and 0.22 m 3 /kgVS fed , respectively). Intermittent mixing improved the retention of methanogens inside the reactor and reduced the washout effect even at low HRT (5% TS). Copyright © 2017 Elsevier Ltd. All rights reserved.

Hydrogen production by hyperthermophilic and extremely thermophilic bacteria and archaea: mechanisms for reductant disposal

NARCIS (Netherlands)

Verhaart, M.R.A.; Bielen, A.A.M.; Oost, van der J.; Stams, A.J.M.; Kengen, S.W.M.

2010-01-01

Hydrogen produced from biomass by bacteria and archaea is an attractive renewable energy source. However, to make its application more feasible, microorganisms are needed with high hydrogen productivities. For several reasons, hyperthermophilic and extremely thermophilic bacteria and archaea are

Microbial community changes in methanogenic granules during the transition from mesophilic to thermophilic conditions

DEFF Research Database (Denmark)

Zhu, Xinyu; Kougias, Panagiotis; Treu, Laura

2017-01-01

Upflow anaerobic sludge blanket (UASB) reactor is one of the most applied technologies for various high-strength wastewater treatments. The present study analysed the microbial community changes in UASB granules during the transition from mesophilic to thermophilic conditions. Dynamicity...

Enrichment of Thermophilic Syntrophic Anaerobic Glutamate-Degrading Consortia using a Dialysis Membrane Reactor

NARCIS (Netherlands)

Plugge, C.M.; Stams, A.J.M.

2002-01-01

A dialysis cultivation system was used to enrich slow-growing moderately thermophilic anaerobic bacteria at high cell densities. Bicarbonate buffered mineral salts medium with 5 mM glutamate as the sole carbon and energy source was used and the incubation temperature was 55 degrees C. The reactor

Distinctive properties of high hydrogen producing extreme thermophiles, Caldicellulosiruptor saccharolyticus and Thermotaga elfii

NARCIS (Netherlands)

Niel, van E.W.J.; Budde, M.A.W.; Haas, de G.G.; Wal, van der F.J.; Claassen, P.A.M.; Stams, A.J.M.

2002-01-01

Growth and hydrogen production by two extreme thermophiles during sugar fermentation was investigated. In cultures of Caldicellulosiruptor saccharolyticus grown on sucrose and Thermotoga elfii grown on glucose stoichiometries of 3.3 mol of hydrogen and 2 mol of acetate per mol C6-sugar unit were

Bioleaching of electronic scrap by mixed culture of moderately thermophilic microorganisms

Science.gov (United States)

Ivǎnuş, D.; ǎnuş, R. C., IV; Cǎlmuc, F.

2010-06-01

A process for the metal recovery from electronic scrap using bacterial leaching was investigated. A mixed culture of moderately thermophilic microorganisms was enriched from acid mine drainages (AMDs) samples collected from several sulphide mines in Romania, and the bioleaching of electronic scrap was conducted both in shake flask and bioreactor. The results show that in the shake flask, the mixture can tolerate 50 g/L scrap after being acclimated to gradually increased concentrations of scrap. The copper extraction increases obviously in bioleaching of scrap with moderately thermophilic microorganisms supplemented with 0.4 g/L yeast extract at 180 r/min, 74% copper can be extracted in the pulp of 50 g/L scrap after 20 d. Compared with copper extractions of mesophilic culture, unacclimated culture and acclimated culture without addition of yeast extract, that of accliniated culture with addition of yeast extract is increased by 53%, 44% and 16%, respectively. In a completely stirred tank reactor, the mass fraction of copper and total iron extraction reach up to 81% and 56%, respectively. The results also indicate that it is necessary to add a large amount of acid to the pulp to extract copper from electronic scrap effectively.

Heterologous expression of cellobiohydrolases in filamentous fungi

DEFF Research Database (Denmark)

Zoglowek, Marta; Lübeck, Peter S.; Ahring, Birgitte K.

2015-01-01

Cellobiohydrolases are among the most important enzymes functioning in the hydrolysis of crystalline cellulose, significantly contributing to the efficient biorefining of recalcitrant lignocellulosic biomass into biofuels and bio-based products. Filamentous fungi are recognized as both well...... into valuable products. However, due to low cellobiohydrolase activities, certain fungi might be deficient with regard to enzymes of value for cellulose conversion, and improving cellobiohydrolase expression in filamentous fungi has proven to be challenging. In this review, we examine the effects of altering...... promoters, signal peptides, culture conditions and host post-translational modifications. For heterologous cellobiohydrolase production in filamentous fungi to become an industrially feasible process, the construction of site-integrating plasmids, development of protease-deficient strains and glycosylation...

Distinguishing ectomycorrhizal and saprophytic fungi using carbon and nitrogen isotopic compositions

Directory of Open Access Journals (Sweden)

Weiguo Hou

2012-05-01

Full Text Available Ectomycorrhizal fungi, a group of widespread symbiotic fungi with plant, obtain carbon source from trees and improve plant mineral nutrient uptake with their widespread hyphal network. Ectomycorrhizal fungi can be used as inoculants to improve the survival rates of plantation. Saprophytic fungi use the nutrition from the debris of plant or animals, and it is difficult to distinguish the saprophytic and ectomycorrhizal fungi by morphological and anatomic methods. In this research, the differences of stable carbon and nitrogen isotopic compositions of these fungi were analyzed. The results showed that the abundances of 13C of were higher than those of ectomycorrhizal fungi and the abundances of 15N of saprophytic fungi were lower than those of ectomycorrhizal fungi. Such differences of stable carbon and nitrogen isotopic compositions between ectomycorrhizal fungi and saprophytic fungi can be ascribed to their different nutrition sources and ecological functions. These results collectively indicate that stable carbon and nitrogen isotopic compositions are an effective proxy for distinguishing between ectomycorrhizal and saprophytic fungi.

Thermal adaptation of mesophilic and thermophilic FtsZ assembly by modulation of the critical concentration.

Directory of Open Access Journals (Sweden)

Luis Concha-Marambio

Full Text Available Cytokinesis is the last stage in the cell cycle. In prokaryotes, the protein FtsZ guides cell constriction by assembling into a contractile ring-shaped structure termed the Z-ring. Constriction of the Z-ring is driven by the GTPase activity of FtsZ that overcomes the energetic barrier between two protein conformations having different propensities to assemble into polymers. FtsZ is found in psychrophilic, mesophilic and thermophilic organisms thereby functioning at temperatures ranging from subzero to >100°C. To gain insight into the functional adaptations enabling assembly of FtsZ in distinct environmental conditions, we analyzed the energetics of FtsZ function from mesophilic Escherichia coli in comparison with FtsZ from thermophilic Methanocaldococcus jannaschii. Presumably, the assembly may be similarly modulated by temperature for both FtsZ orthologs. The temperature dependence of the first-order rates of nucleotide hydrolysis and of polymer disassembly, indicated an entropy-driven destabilization of the FtsZ-GTP intermediate. This destabilization was true for both mesophilic and thermophilic FtsZ, reflecting a conserved mechanism of disassembly. From the temperature dependence of the critical concentrations for polymerization, we detected a change of opposite sign in the heat capacity, that was partially explained by the specific changes in the solvent-accessible surface area between the free and polymerized states of FtsZ. At the physiological temperature, the assembly of both FtsZ orthologs was found to be driven by a small positive entropy. In contrast, the assembly occurred with a negative enthalpy for mesophilic FtsZ and with a positive enthalpy for thermophilic FtsZ. Notably, the assembly of both FtsZ orthologs is characterized by a critical concentration of similar value (1-2 μM at the environmental temperatures of their host organisms. These findings suggest a simple but robust mechanism of adaptation of FtsZ, previously shown

Mesophilic and thermophilic anaerobic co-digestion of rendering plant and slaughterhouse wastes.

Science.gov (United States)

Bayr, Suvi; Rantanen, Marianne; Kaparaju, Prasad; Rintala, Jukka

2012-01-01

Co-digestion of rendering and slaughterhouse wastes was studied in laboratory scale semi-continuously fed continuously stirred tank reactors (CSTRs) at 35 and 55 °C. All in all, 10 different rendering plant and slaughterhouse waste fractions were characterised showing high contents of lipids and proteins, and methane potentials of 262-572 dm(3)CH(4)/kg volatile solids(VS)(added). In mesophilic CSTR methane yields of ca 720 dm(3) CH(4)/kg VS(fed) were obtained with organic loading rates (OLR) of 1.0 and 1.5 kg VS/m(3) d, and hydraulic retention time (HRT) of 50 d. For thermophilic process, the lowest studied OLR of 1.5 kg VS/m(3) d, turned to be unstable after operation of 1.5 HRT, due to accumulating ammonia, volatile fatty acids (VFAs) and probably also long chain fatty acids (LCFAs). In conclusion, mesophilic process was found to be more feasible for co-digestion than thermophilic process, methane yields being higher and process more stable in mesophilic conditions. Copyright © 2011 Elsevier Ltd. All rights reserved.

Thermophilic enzymes and their applications in biocatalysis: a robust aldo-keto reductase.

Science.gov (United States)

Willies, Simon; Isupov, Misha; Littlechild, Jennifer

2010-09-01

Extremophiles are providing a good source of novel robust enzymes for use in biocatalysis for the synthesis of new drugs. This is particularly true for the enzymes from thermophilic organisms which are more robust than their mesophilic counterparts to the conditions required for industrial bio-processes. This paper describes a new aldo-keto reductase enzyme from a thermophilic eubacteria, Thermotoga maritima which can be used for the production of primary alcohols. The enzyme has been cloned and over-expressed in Escherichia coli and has been purified and subjected to full biochemical characterization. The aldo-keto reductase can be used for production of primary alcohols using substrates including benzaldehyde, 1,2,3,6-tetrahydrobenzaldehyde and para-anisaldehyde. It is stable up to 80 degrees C, retaining over 60% activity for 5 hours at this temperature. The enzyme at pH 6.5 showed a preference for the forward, carbonyl reduction. The enzyme showed moderate stability with organic solvents, and retained 70% activity in 20% (v/v) isopropanol or DMSO. These properties are favourable for its potential industrial applications.

Bacterial community analysis of swine manure treated with autothermal thermophilic aerobic digestion.

Science.gov (United States)

Han, Il; Congeevaram, Shankar; Ki, Dong-Won; Oh, Byoung-Taek; Park, Joonhong

2011-02-01

Due to the environmental problems associated with disposal of livestock sludge, many stabilization studies emphasizing on the sludge volume reduction were performed. However, little is known about the microbial risk present in sludge and its stabilized products. This study microbiologically explored the effects of anaerobic lagoon fermentation (ALF) and autothermal thermophilic aerobic digestion (ATAD) on pathogen-related risk of raw swine manure by using culture-independent 16S rDNA cloning and sequencing methods. In raw swine manure, clones closely related to pathogens such as Dialister pneumosintes, Erysipelothrix rhusiopathiae, Succinivibrioan dextrinosolvens, and Schineria sp. were detected. Meanwhile, in the mesophilic ALF-treated swine manure, bacterial community clones closely related to pathogens such as Schineria sp. and Succinivibrio dextrinosolvens were still detected. Interestingly, the ATAD treatment resulted in no detection of clones closely related to pathogens in the stabilized thermophilic bacterial community, with the predominance of novel Clostridia class populations. These findings support the superiority of ATAD in selectively reducing potential human and animal pathogens compared to ALF, which is a typical manure stabilization method used in livestock farms.

Common wood decay fungi found in the Caribbean Basin

Science.gov (United States)

D. Jean. Lodge

2016-01-01

There are hundreds of wood-decay fungi in the Caribbean Basin, but relatively few of these are likely to grow on manmade structures built of wood or wood-composites. The wood-decay fungi of greatest concern are those that cause brown-rot, and especially brown-rot fungi that are resistant to copper-based wood preservatives. Some fungi that grow in the Caribbean and...

Genome Studies on Nematophagous and Entomogenous Fungi in China

Science.gov (United States)

Zhang, Weiwei; Cheng, Xiaoli; Liu, Xingzhong; Xiang, Meichun

2016-01-01

The nematophagous and entomogenous fungi are natural enemies of nematodes and insects and have been utilized by humans to control agricultural and forestry pests. Some of these fungi have been or are being developed as biological control agents in China and worldwide. Several important nematophagous and entomogenous fungi, including nematode-trapping fungi (Arthrobotrys oligospora and Drechslerella stenobrocha), nematode endoparasite (Hirsutella minnesotensis), insect pathogens (Beauveria bassiana and Metarhizium spp.) and Chinese medicinal fungi (Ophiocordyceps sinensis and Cordyceps militaris), have been genome sequenced and extensively analyzed in China. The biology, evolution, and pharmaceutical application of these fungi and their interacting with host nematodes and insects revealed by genomes, comparing genomes coupled with transcriptomes are summarized and reviewed in this paper. PMID:29376926

Fossil evidence of the zygomycetous fungi

NARCIS (Netherlands)

Krings, M.; Taylor, T.N.; Dotzler, N.

2013-01-01

Molecular clock data indicate that the first zygomycetous fungi occurred on Earth during the Precambrian, however, fossil evidence of these organisms has been slow to accumulate. In this paper, the fossil record of the zygomycetous fungi is compiled, with a focus on structurally preserved

Foliar fungi of Scots pine (Pinus sylvestris)

OpenAIRE

Millberg, Hanna

2015-01-01

Scots pine (Pinus sylvestris) is an ecologically and economically important tree species in Fennoscandia. Scots pine needles host a variety of fungi, some with the potential to profoundly influence their host. These fungi can have beneficial or detrimental effects with important implications for both forest health and primary production. In this thesis, the foliar fungi of Scots pine needles were investigated with the aim of exploring spatial and temporal patterns, and development with needle...