Home About us Contact
|
Home About us Contact | ||
|
|
||
Thermophilus
Kinds of Thermophilus Selected AbstractsAnalyzing and monitoring of phage,bacteria interaction using CEELECTROPHORESIS, Issue 20 2009Esra Acar Soykut Abstract The utilization of CE for monitoring bacteria,phage interaction was investigated in this study. Streptococcus thermophilus and Lactobacillus bulgaricus strains and their phages were used as model bacteria and phages for the purpose of validation in this study. CE with heterogeneous polymer polyethylene oxide was utilized for the separation of intact bacteria and investigation of phage,bacteria interaction. An intact phage detection was carried out with CZE by adding SDS in the running buffer. Calibration graphs of bacteria and phages were obtained with R2 values of 0.963 and 0.937, respectively. S. thermophilus strain was infected with its virulent phage B3-X18 for investigation of phage,bacteria interaction. It was observed in capillary electropherogram that the culture was lysed depending on the multiplicity of infection value and it showed to be completely lysed when the multiplicity of infection value was 10. The interaction of S. thermophilus strain with L. bulgaricus phage was also investigated by using a CE and a microbiological method and it was observed that the L. bulgaricus phage attached itself to the cell wall of S. thermophilus strain without damaging the cell. [source] The role of the nitrate respiration element of Thermus thermophilus in the control and activity of the denitrification apparatusENVIRONMENTAL MICROBIOLOGY, Issue 2 2008Felipe Cava Summary The nitrate conjugative element (NCE) encodes the ability to respire nitrate in the facultative Thermus thermophilus NAR1 strain. This process is carried out by two heterotetrameric enzymes that catalyse the oxidation of NADH (Nrc) and the reduction of nitrate (Nar), whose expression is activated by the NCE-encoded transcription factors DnrS and DnrT. We report the presence of NCE in other facultative strains of T. thermophilus and analyse its role in subsequent steps of the denitrification pathway. We encountered that nrc mutants of denitrifying strains show a decrease in anaerobic growth rates not only with nitrate, but also with nitrite, NO and N2O, which is concomitant to their lower NADH oxidation activities in vitro. We show that nitrate, nitrite and NO are activating signals for transcription of nrc in these strains. Finally, we demonstrate that DnrS and DnrT are required for anaerobic growth not only with nitrate, but also with nitrite, NO and N2O. These data allow us to conclude that: (i) Nrc constitutes the main electron donor for the four reductases of the denitrification pathway, and (ii) the NCE controls the expression of the whole denitrification pathway and the repression of the aerobic respiration through the transcription factors DnrS and DnrT. [source] Thermus thermophilus Glycosynthases for the Efficient Synthesis of Galactosyl and Glucosyl ,-(1,3) - GlycosidesEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 10 2005Jullien Drone Abstract Inverting mutant glycosynthases were designed according to the Withers strategy, starting from wild-type Thermus thermophilus retaining Tt-,-Gly glycosidase. Directed mutagenesis of catalytic nucleophile glutamate 338 by alanine, serine, and glycine afforded the E338A, E338S, and E338G mutant enzymes, respectively. As was to be expected, the mutants were unable to catalyze the hydrolysis of the transglycosidation products. In agreement with previous results, the E338S and E338G catalysts were much more efficient than E338A. Moreover, our results showed that these enzymes were inactive in the hydrolysis of the ,- D -glycopyranosyl fluorides used as donors, and so suitable experimental conditions, under which the rate of spontaneous hydrolysis of the donor was considerably lower than that of enzymatic transglycosidation, provided galactosyl and glucosyl ,-(1,3) - glycosides in yields of up to 90,%. The structure of native Tt-,-Gly available in the Protein Data Bank offers a good basis for interpretation of our results by means of molecular modeling. Thus, in the case of the E338S mutant, a lower energy of the system was obtained when the donor and the acceptor were in the right position to form the ,-(1,3) - glycosidic bond. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2005) [source] Effect of the lactic acid bacterium Streptococcus thermophilus on stratum corneum ceramide levels and signs and symptoms of atopic dermatitis patientsEXPERIMENTAL DERMATOLOGY, Issue 5 2003Luisa Di Marzio Abstract:, A reduced amount of total ceramides could be responsible for functional abnormalities of the skin of atopic dermatitis (AD) patients. The ability of an experimental cream containing sonicated Streptococcus thermophilus to increase skin ceramide levels in healthy subjects has been previously reported. The aim of the present work was to investigate the effects of the topical administration of a S. thermophilus -containing cream on ceramide levels of stratum corneum from AD patients. A 2-week application of the cream, containing a sonicated preparation of the lactic acid bacterium S. thermophilus, in the forearm skin of 11 patients led to a significant and relevant increase of skin ceramide amounts, which could have resulted from the sphingomyelin hydrolysis through the bacterial sphingomyelinase. Moreover, in all patients the topical application of our experimental cream also resulted in the improvement of the signs and symptoms characteristic of AD skin (i.e. erythema, scaling, pruritus). [source] Purification of three aminotransferases from Hydrogenobacter thermophilus TK-6 , novel types of alanine or glycine aminotransferaseFEBS JOURNAL, Issue 8 2010Enzymes, catalysis Aminotransferases catalyse synthetic and degradative reactions of amino acids, and serve as a key linkage between central carbon and nitrogen metabolism in most organisms. In this study, three aminotransferases (AT1, AT2 and AT3) were purified and characterized from Hydrogenobacter thermophilus, a hydrogen-oxidizing chemolithoautotrophic bacterium, which has been reported to possess unique features in its carbon and nitrogen anabolism. AT1, AT2 and AT3 exhibited glutamate:oxaloacetate aminotransferase, glutamate:pyruvate aminotransferase and alanine:glyoxylate aminotransferase activities, respectively. In addition, both AT1 and AT2 catalysed a glutamate:glyoxylate aminotransferase reaction. Interestingly, phylogenetic analysis showed that AT2 belongs to aminotransferase family IV, whereas known glutamate:pyruvate aminotransferases and glutamate:glyoxylate aminotransferases are members of family I,. In contrast, AT3 was classified into family I, distant from eukaryotic alanine:glyoxylate aminotransferases which belong to family IV. Although Thermococcus litoralis alanine:glyoxylate aminotransferase is the sole known example of family I alanine:glyoxylate aminotransferases, it is indicated that this alanine:glyoxylate aminotransferase and AT3 are derived from distinct lineages within family I, because neither high sequence similarity nor putative substrate-binding residues are shared by these two enzymes. To our knowledge, this study is the first report of the primary structure of bacterial glutamate:glyoxylate aminotransferase and alanine:glyoxylate aminotransferase, and demonstrates the presence of novel types of aminotransferase phylogenetically distinct from known eukaryotic and archaeal isozymes. [source] High thermal and chemical stability of Thermus thermophilus seven-iron ferredoxinFEBS JOURNAL, Issue 23 2003Linear clusters form at high pH on polypeptide unfolding To probe the stability of the seven-iron ferredoxin from Thermus thermophilus (FdTt), we investigated its chemical and thermal denaturation processes in solution. As predicted from the crystal structure, FdTt is extremely resistant to perturbation. The guanidine hydrochloride-induced unfolding transition shows a midpoint at 6.5 m (pH 7, 20 °C), and the thermal midpoint is above boiling, at 114 °C. The stability of FdTt is much lower at acidic pH, suggesting that electrostatic interactions are important for the high stability at higher pH. On FdTt unfolding at alkaline pH, new absorption bands at 520 nm and 610 nm appear transiently, resulting from rearrangement of the cubic clusters into linear three-iron species. A range of iron,sulfur proteins has been found to accommodate these novel clusters in vitro, although no biological function has yet been assigned. [source] tmRNA from Thermus thermophilusFEBS JOURNAL, Issue 3 2003Interaction with alanyl-tRNA synthetase, elongation factor Tu The interaction of a Thermus thermophilus tmRNA transcript with alanyl-tRNA synthetase and elongation factor Tu has been studied. The synthetic tmRNA was found to be stable up to 70 °C. The thermal optimum of tmRNA alanylation was determined to be around 50 °C. At 50 °C, tmRNA transcript was aminoacylated by alanyl-tRNA synthetase with 5.9 times lower efficiency (kcat/Km) than tRNAAla, primarily because of the difference in turnover numbers (kcat). Studies on EF-Tu protection of Ala,tmRNA against alkaline hydrolysis revealed the existence of at least two different binding sites for EF-Tu on charged tmRNA. The possible nature of these binding sites is discussed. [source] Shuffling genes around in hot environments: the unique DNA transporter of Thermus thermophilusFEMS MICROBIOLOGY REVIEWS, Issue 3 2009Beate Averhoff Abstract Natural transformation permits the transport of DNA through bacterial membranes and represents a dominant mode for the transfer of genetic information between bacteria and between microorganisms of distant evolutionary lineages and even between members of different domains. This phenomenon, known as horizontal, or lateral, gene transfer, has been a major force for genome plasticity over evolutionary history, and is largely responsible for the spread of fitness-enhancing traits, including antibiotic resistance and virulence factors. In particular, for adaptation of prokaryotes to extreme environments, lateral gene transfer seems to have played a crucial role. Here, we present a survey of the natural transformation machinery of the thermophile Thermus thermophilus HB27. A tentative model of the transformation machinery comprising of components similar to proteins of type IV pili and type II secretion systems is presented. A comparative discussion of the subunits and the structure of the DNA translocator and the underlying mechanism of transfer of free DNA in T. thermophilus highlights conserved and unique features of the DNA translocator in T. thermophilus. We hypothesize that the extraordinary broad substrate specificity and the high efficiency of the T. thermophilus DNA uptake system is of major importance for thermoadaptation and interdomain DNA transfer in hot environments. [source] Effect of fat and non-fat dry matter of milk, and starter type, on the rheological properties of set during the coagulation processINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 3 2007Özlem Gün Summary The effect of fat and non-fat dry matter (NFDM) of milk, and starter type, on the rheological properties of set yoghurt during the coagulation process was studied using a rotational viscosimeter. The pH was monitored simultaneously with viscosity readings. Recombined milk was cultured with yoghurt starter culture (YC), consisting of Streptococcus thermophilus and Lactobacillus bulgaricus, or ABT1, consisting of S. thermophilus, Lactobacillus acidophilus and bifidobacteria, at 40 °C until the pH dropped to 4.7. Yoghurt made with ABT1 had higher viscosity values than yoghurt made with YC. The NFDM content of milk had a major effect on the viscosity of curd made using YC or ABT1. For YC and ABT1, three stages were identified on the viscosity,incubation time curves and viscosity behaviour was different in each stage. [source] Transcriptional analysis of the gdhA gene in Streptococcus thermophilusJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2009C. Lazzi Abstract Aims:, To study the transcriptional analysis of glutamate dehydrogenase gene, involved in the amino acid conversion to aroma compound in Streptococcus thermophilus. Methods and Results:, Analysis of the gdhA gene nucleotide sequence of S. thermophilus CNRZ1066 revealed that the coding region is 1353 nucleotides long. The deduced amino acids sequence exhibits the putative GDH active site and some conserved domains characteristic of family I of hexameric GDHs. Phylogenetic analysis revealed that the gdh gene of S. thermophilus clustered with the orthologues of other streptococci such as Streptococcus mutans, Streptococcus agalactiae and Streptococcus infantarius. Studying the structural organization of the gdhA locus the amino acid similarity of GDHs was higher than 87%, but the locus organization was not conserved. A dominant transcript of approximately 1·4 kbp was revealed by Northern blot hybridization, suggesting that gdhA mRNA is monocystronic. Primer extension showed that transcription start point of gdhA was localized 43 bp upstream of the potential start codon (ATG). Conclusions:, The gdhA represents a monocistronic operon highly conserved in phylogenetic-related bacteria. Significance and Impact of the Study:, A deeper knowledge of gdh transcriptional mechanisms could lead to develop S. thermophilus industrial starter cultures with optimized aromatic properties. [source] Characterization of dominant microbiota of a Ghanaian fermented milk product, nyarmie, by culture- and nonculture-based methodsJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2006M. Obodai Abstract Aims:, To characterize the predominant micro-organisms in a Ghanaian traditional fermented dairy product, nyarmie, made from cows' milk, using both culture- and nonculture-based methods. Methods and Results:, Samples of nyarmie were analysed from three production sites in Accra, by determining the counts on selective culture media. The microbial diversity occurring in nyarmie was also evaluated by 16S/18S ribosomal DNA PCR amplification and denaturing gradient gel electrophoresis. Results showed that nyarmie contained lactococci and lactobacilli in the range of 108 and 1010 CFU ml,1, respectively, and yeasts at around 107 CFU ml,1. The pH ranged between 3·49 and 4·25. The predominant lactic acid bacteria (LAB) in nyarmie were Leuconostocmesenteroides ssp. mesenteroides, Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lact.helveticus, Lact. delbrueckii ssp. lactis and Lactococcus lactis, while Saccharomyces cerevisiae was the predominant yeast species. Lactobacillus delbrueckii ssp. delbrueckii was not detected by cultivation but its predominance was revealed by PCR-DGGE analysis. Conclusions:, The flora in products from different producers varied in the LAB composition present and may result in variations in product quality. Significance and Impact of the Study:, Development and use of starter cultures for nyarmie may be beneficial in improving the consistency of product quality. [source] Autolytic phenotype of Lactococcus lactis strains isolated from traditional Tunisian dairy productsJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2002H. Ouzari Aims:,To evaluate the autolytic properties of Lactococcus lactis strains isolated from artisan Tunisian dairy products, their peptidoglycan hydrolase content and their activity spectrum. Methods and Results:,The autolytic phenotype of Lactococcus strains was evaluated under starvation conditions in potassium phosphate buffer. The results obtained highlighted a high degree of diversity among the strains analysed, allowing the identification of high and low autolytic Lactococcus lactis strains. Peptidoglycan hydrolase content was evaluated by renaturing SDS-PAGE using cells of Micrococcus lysodeikticus as a target for the enzymatic activity. A major activity band migrating at about 45 kDa was observed. The lytic activity, evaluated in the presence of different chemicals, was retained in 8% NaCl, 15 mmol l,1 CaCl2, and in a pH range between 5 and 9·5. The substrate specificity of peptidoglycan hydrolase from Lactococcus strains was evaluated in renaturing SDS-PAGE incorporating cells of different bacterial species. The major autolysin of Lactococcus lactis was active against cells of Lactococcus lactis subsp. lactis, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus and Listeria monocytogenes. Conclusions:,Autolytic activity is widely distributed in Lactococcus lactis and the rate of autolysis is strain-dependent. The major peptidoglycan hydrolase showed a wide spectrum of activity against several lactic acid bacteria and bacterial species involved in food-related infection. Significance and Impact of the Study:,The autolytic phenotype of Lactococcus lactis strains isolated from Tunisian artisan dairy products has been determined, and the data obtained should allow the selection of strains of technological interest in the cheese-ripening process. [source] ANTIBACTERIAL ACTIVITY AND CHEMICAL CONSTITUTIONS OF OLEA EUROPAEA L. LEAF EXTRACTSJOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 3 2010MIHRIBAN KORUKLUOGLU ABSTRACT The in vitro antimicrobial activity of aqueous, acetone, diethyl ether and ethyl alcohol extracts of olive leaves (Olea europaea L.) was studied. The aqueous extract of olive leaves had no antibacterial effect against the test microorganisms, whereas acetone extract showed inhibitory effect on Salmonella enteritidis, Bacillus cereus, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Streptococcus thermophilus and Lactobacillus bulgaricus. Furthermore, the antimicrobial activities of some phenolic compounds against microorganisms were tested. The most effective compound was found to be oleuropein while syringic acid was found ineffective. The characterization of phenolic compounds in different extracts determined by high performance liquid chromatography-air pressure chemical ionization-mass spectrometry detector (HPLC-APCI-MSD GC-MS) gas chromatography-mass spectrometry (GC-MS). The acetone and the ethyl alcohol extracts had the most and the least oleuropein content, respectively. PRACTICAL APPLICATIONS In recent years the extracts of many plant species have become popular, and attempts to characterize their bioactive principles have gained speed for many pharmaceutical and food-processing applications. Especially, antimicrobial properties of plants have revived as a consequence of current problems associated with the use of chemical preservatives. Because of consumers' negative perspectives of synthetic preservatives, attention is shifting toward natural alternatives. The findings suggest that olive leaf extracts and their phenolic compounds have good potential as antibacterial substances in food preservation as they may be more acceptable to consumers and the regulatory agencies in comparison with synthetic chemical compounds. [source] EFFECT OF THERMOPHILIC LACTIC ACID BACTERIA ON THE FATE OF ENTEROBACTER SAKAZAKII DURING PROCESSING AND STORAGE OF PLAIN YOGURTJOURNAL OF FOOD SAFETY, Issue 2 2008REYAD R. SHAKER ABSTRACT Survival and growth of Enterobacter sakazakii during processing and storage of plain yogurt were investigated. Preheated rehydrated milk was inoculated with a cocktail culture of E. sakazakii (103 cfu/mL of milk) and/or with thermophilic yogurt starter culture of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus. The inoculated milk was incubated at 40C for 5 h, then the samples were cooled and subsequently stored at 4C for up to 7 days. The results showed that E. sakazakii grew at an early stage of fermentation but declined at the end of the process. There was no significant difference between the populations of E. sakazakii in the presence or absence of lactic acid bacteria during the first 4 h of the incubation period but there was significant difference during the last hour of the incubation period. The populations of E. sakazakii decreased significantly during cooling and storage of yogurt (pH 4.2,4.7) compared with nonfermented milk samples at 4C. The presence of E. sakazakii did not have a significant effect on the growth of LAB during fermentation and storage of yogurt. The results obtained from this study indicate that the pH of yogurt and storage temperature were critical to the survival and growth of E. sakazakii in the manufacture of plain yogurt. PRACTICAL APPLICATIONS Enterobacter sakazakii prevalence in milk products and the production environment has been documented. The results obtained from this study may be of use to dairy producers to manufacture safe products using thermophilic lactic acid bacteria. These bacteria decreased the pH of milk in less than 5 h, resulting in E. sakazakii reduction. pH of yogurt during the fermentation process is a critical control point that should be monitored to produce safe products. [source] Viability During Storage of Selected Probiotic Lactobacilli and Bifidobacteria in a Yogurt-like ProductJOURNAL OF FOOD SCIENCE, Issue 8 2002S.E. Gilliland ABSTRACT: Multiple species cultures, including 2 strains of Streptococcus thermophilus and Lactobacillus acidophilus NCFM plus 1 strain each of Bifidobacterium longum and Lactobacillus casei, were used to make yogurt-like products. The lactobacilli and bifidobacteria were tested for growth in the products and subsequent viability during refrigerated storage. During fermentation, L. casei Com-5 actually declined in numbers, while L. casei E5 and E10 increased about 2 fold. Numbers of B. longum S9 increased about 3 fold while B. longum Com-4 did not increase. During storage, L. acidophilus NCFM appeared stable in all mixtures and both strains of bifidobacteria decreased. Lactobacillus casei E5 and E10 were more stable than was L. casei Com-5. [source] 2,,3,-Cyclic nucleotide 3,-phosphodiesterase: A novel RNA-binding protein that inhibits protein synthesisJOURNAL OF NEUROSCIENCE RESEARCH, Issue 5 2009Michel Gravel Abstract 2,,3,-Cyclic nucleotide 3,-phosphodiesterase (CNP) is one of the earliest myelin-related proteins to be specifically expressed in differentiating oligodendrocytes (ODCs) in the central nervous system (CNS) and is implicated in myelin biogenesis. CNP possesses an in vitro enzymatic activity, whose in vivo relevance remains to be defined, because substrates with 2,,3,-cyclic termini have not yet been identified. To characterize CNP function better, we previously determined the structure of the CNP catalytic domain by NMR. Interestingly, the structure is remarkably similar to the plant cyclic nucleotide phosphodiesterase (CPDase) from A. thaliana and the bacterial 2,-5, RNA ligase from T. thermophilus, which are known to play roles in RNA metabolism. Here we show that CNP is an RNA-binding protein. Furthermore, by using precipitation analyses, we demonstrate that CNP associates with poly(A)+ mRNAs in vivo and suppresses translation in vitro in a dose-dependent manner. With SELEX, we isolated RNA aptamers that can suppress the inhibitory effect of CNP on translation. We also demonstrate that CNP1 can bridge an association between tubulin and RNA. These results suggest that CNP1 may regulate expression of mRNAs in ODCs of the CNS. © 2008 Wiley-Liss, Inc. [source] RHEOLOGY AND TEXTURE OF SET YOGURT AS AFFECTED BY INULIN ADDITIONJOURNAL OF TEXTURE STUDIES, Issue 6 2008TATDAO PASEEPHOL ABSTRACT The effect of inulin addition on rheology and texture of nonfat yogurt (NFY) was studied by the large (texture analyzer) and small (dynamic oscillatory rheometry) deformation tests. Skim milks (12% solids) were supplemented with three commercial chicory inulins with different chain lengths at 4% level and inoculated with mixed cultures of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus. Textural and rheological tests were conducted after an overnight storage at 4C and during a shelf life of 28 days. A good correlation was observed between the two tests. During storage, all inulin-containing yogurts in comparison with the control NFY were characterized by lower values of firmness, apparent viscosity, yield stress, complex viscosity, storage modulus and loss modulus. NFYs supplemented with long-chain inulin demonstrated a rheological behavior closer to that of control full-fat yogurt. PRACTICAL APPLICATIONS The use of inulin and oligofructose in the food industry has grown recently because of their interesting nutritional and technological properties. In the work presented here, three commercial inulin products with different chemical composition and functions were added to yogurt milk, and changes in rheological properties of yogurt were investigated in comparison with nonsupplemented control yogurts. According to the manufacturer's specifications, Raftiline HP and Raftiline GR are suitable for fat replacement because of the presence of long-chain inulins, while Raftilose P95, being rich in mono- and disaccharides, is highly recommended for sugar replacement. The knowledge obtained from this study may be applied for the development of nonfat yogurts to satisfy the nutritional expectations of ever-increasing health conscious consumers and at the same time to match the sensory and textural characteristics of full-fat yogurt. [source] EFFECT OF COLD STORAGE ON CULTURE VIABILITY AND SOME RHEOLOGICAL PROPERTIES OF FERMENTED MILK PREPARED WITH YOGURT AND PROBIOTIC BACTERIAJOURNAL OF TEXTURE STUDIES, Issue 1 2008MARIA REGINA DAMIN ABSTRACT We examined the effect of storage time on culture viability and some rheological properties (yield stress, storage modulus, loss modulus, linear viscoelastic region, structural recuperation and firmness) of fermented milk made with Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus (LA) and Bifidobacterium animalis ssp. lactis in coculture with Streptococcus thermophilus (ST). Acidification profiles and factors that affect viability (postfermentation acidification, acidity and dissolved oxygen) were also studied during 35 days at 4C. Fermented milk prepared with a coculture of ST and Bifidobacterium lactis gave the most constant rheological behavior and the best cell viability during cold storage; it was superior to ST plus LA for probiotic fermented milk production. PRACTICAL APPLICATIONS Probiotic cultures should grow quickly in milk, provide adequate sensory and rheological properties to the product, and remain viable during storage. Commercially, it is very common to use yogurt starter culture (i.e. Streptococcus thermophilus[ST] and Lactobacillus delbrueckii ssp. bulgaricus) in combination with the probiotic bacteria in order to reduce fermentation time. However, LB tends to post acidify fermented milk, which reduces the viability of the probiotic bacteria; thus, it is recommended to use starter cultures devoid of this species. We found that the technological properties and the viability of the probiotic bacterium Bifidobacterium animalis ssp. lactis BL O4 in coculture with ST make it suitable for probiotic fermented milk production; it produces rheological characteristics similar to those of yogurt. [source] Conjugated linoleic acid conversion by dairy bacteria cultured in MRS broth and buffalo milkLETTERS IN APPLIED MICROBIOLOGY, Issue 5 2007C.P. Van Nieuwenhove Abstract Aims:, To evaluate strains of Lactobacilli, Bifidobacteria and Streptococci for their ability to produce conjugated linoleic acid (CLA) from free linoleic acid (LA). Methods and Results:, Eight dairy bacteria tolerant to LA were grown in MRS broth containing LA (200 ,g ml,1) and CLA was assessed. Seven bacteria were able to form CLA after 24 h of incubation, varying percentage conversion between 17% and 36%. Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium bifidum and Streptococcus thermophilus showed the highest LA conversion and were inoculated into buffalo milk supplemented with different concentration of LA. The production of CLA at 200 ,g ml,1 of LA was two- or threefold in milk than MRS broth. All evaluated strains were able to produce CLA from high LA levels (1000 ,g ml,1). Conclusions:, The most tolerant strain to LA was Lact. casei. Lacttobacillus rhamnosus produced the maximum level of CLA at high LA concentrations (800 ,g ml,1). The selected bacteria may be considered as adjunct cultures to be included on dairy fermented products manufacture. Low concentration of LA must be added to the medium to enhance CLA formation. Significance and Impact of the Study:, The production of CLA by strains using milks from regional farms as medium offer a possible mechanism to enhance this beneficial compound in dairy products and those the possibility to develop functional foods. [source] Regulation of branched-chain amino acid biosynthesis by ,-acetolactate decarboxylase in Streptococcus thermophilusLETTERS IN APPLIED MICROBIOLOGY, Issue 6 2003C. Monnet Abstract Aims: To demonstrate the presence of an active , -acetolactate decarboxylase in Streptococcus thermophilus and to investigate its physiological function. Methods and Results:Streptococcus thermophilus CNRZ385 contains a gene encoding an , -acetolactate decarboxylase. Comparison of the production of , -acetolactate and its decarboxylation products, by the parent strain and an , -acetolactate decarboxylase-deficient mutant, demonstrated the presence of a control of the pool of ,-acetolactate by valine, leucine and isoleucine. This control occurs via an allosteric activation of the , -acetolactate decarboxylase. Cell-free extracts of S. thermophilus were not able to decarboxylate the isoleucine precursor , -acetohydroxybutyrate. Conclusions: These results strongly suggest that one of the physiological functions of the , -acetolactate decarboxylase in S. thermophilus is to regulate leucine and valine biosynthesis by diverting the flux of , -acetolactate towards acetoin when the branched-chain amino acids are present at a high concentration. Significance and Impact of the Study: Regulation of branched-chain amino acid biosynthesis by , -acetolactate decarboxylase may occur in several other micro-organisms and explain some of their growth properties. [source] Inducible and constitutive expression using new plasmid and integrative expression vectors for Thermus sp.LETTERS IN APPLIED MICROBIOLOGY, Issue 6 2001K.J. Kayser Aims: To develop molecular tools and examine inducible and constitutive gene expression in Thermus thermophilus. Methods and Results: Two plasmid promoter probe vectors and an integrative promoter probe vector were constructed using a promoterless thermostable kanamycin nucleotidyltransferase (KmR) cassette. Three expression vectors were constructed based on a constitutive promoter J17, that functions in both Thermus and Escherichia coli. An inducible expression vector was constructed using the heat-shock inducible promoter (70 to 85°C) from the dnaK gene of T. flavus, and the malate dehydrogenase gene (mdh) from T. flavus was cloned and expressed in both E. coli and T. thermophilus HB27. Conclusions: This report describes the construction and use of improved promoter probe and expression vectors for use in Thermus species. The mdh gene can be used as a high temperature (85°C) reporter gene for Thermus sp. The dnaK promoter is thermo-inducible. Significance and Impact of the Study: The expression vectors and molecular tools described here are significant improvements over previously reported vectors for Thermus sp. The mdh gene and the thermo-inducible dnaK promoter will facilitate high temperature studies employing Thermus species. [source] Cation/proton antiporter complements of bacteria: why so large and diverse?MOLECULAR MICROBIOLOGY, Issue 2 2009Terry A. Krulwich Summary Most bacterial genomes have five to nine distinct genes predicted to encode transporters that exchange cytoplasmic Na+ and/or K+ for H+ from outside the cell, i.e. monovalent cation/proton antiporters. By contrast, pathogens that live primarily inside host cells usually possess zero to one such antiporter while other stress-exposed bacteria exhibit even higher numbers. The monovalent cation/proton antiporters encoded by these diverse genes fall into at least eight different transporter protein families based on sequence similarity. They enable bacteria to meet challenges of high or fluctuating pH, salt, temperature or osmolarity, but we lack explanations for why so many antiporters are needed and for the value added by specific antiporter types in specific settings. In this issue of Molecular Microbiology, analyses of the pH dependence of cytoplasmic [Na+], [K+], pH and transmembrane electrical potential in the ,poly extremophile'Natranaerobius thermophilus are the context for assessment of the catalytic properties of 12 predicted monovalent cation/proton antiporters in the genome of this thermophilic haloalkaliphile. The results provide a profile of adaptations of the poly extremophilic anaerobe, including a proposed role of cytoplasmic buffering capacity. They also provide new perspectives on two large monovalent cation/proton antiporter families, the NhaC and the cation/proton antiporter-3 antiporter families. [source] The CHAP domain of Cse functions as an endopeptidase that acts at mature septa to promote Streptococcus thermophilus cell separationMOLECULAR MICROBIOLOGY, Issue 5 2009Séverine Layec Summary Cell separation is dependent on cell wall hydrolases that cleave the peptidoglycan shared between daughter cells. In Streptococcus thermophilus, this step is performed by the Cse protein whose depletion resulted in the formation of extremely long chains of cells. Cse, a natural chimeric enzyme created by domain shuffling, carries at least two important domains for its activity: the LysM expected to be responsible for the cell wall-binding and the CHAP domain predicted to contain the active centre. Accordingly, the localization of Cse on S. thermophilus cell surface has been undertaken by immunogold electron and immunofluorescence microscopies using of antibodies raised against the N-terminal end of this protein. Immunolocalization shows the presence of the Cse protein at mature septa. Moreover, the CHAP domain of Cse exhibits a cell wall lytic activity in zymograms performed with cell walls of Micrococcus lysodeikticus, Bacillus subtilis and S. thermophilus. Additionally, RP-HPLC analysis of muropeptides released from B. subtilis and S. thermophilus cell wall after digestion with the CHAP domain shows that Cse is an endopeptidase. Altogether, these results suggest that Cse is a cell wall hydrolase involved in daughter cell separation of S. thermophilus. [source] Influence of heat impact in reconstituted skim milk on the properties of yoghurt fermented by ropy or non-ropy starter culturesMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 5 2003Peter Chr. Abstract The paper describes studies on the influence of heat impact in reconstituted skim milk on chemical and functional properties of yoghurt products. Reconstituted skim milk was heated for 20 min at 85°C, 90°C, or 95°C. Ropy (producing exopolysaccharides, EPS) or non-ropy strains of S. thermophilus and L. delbrueckii subsp. bulgaricus were used as starter culture for yoghurt manufacture. The studies have shown that the fermentation times decreased with increasing heat impact when the ropy starter culture was used, while they remained to a far extent unchanged if the non-ropy starter culture was applied. The lactic acid contents of the yoghurt products were in the same range when the milk was heated at 85°C or 90°C, while they were different when milk was heated at 95°C. There was a tendency visible that an increase in preheating leads to increased L(+)- and decreased D(,)-lactic acid contents if the non-ropy culture was applied. Using the ropy culture, it was vice versa. A slightly decrease in proteolysis with increasing heat impact was to be noted with both starter cultures. Concerning the relation of proteolysis to acidification, the fermentation process could be subdivided into three sections with different slopes if the non-ropy starter culture was used, while a linear relation was found if the ropy starter culture was applied. Regarding final product characteristics it was found that the functional properties of yoghurt decreased with increasing heat impact when the ropy starter culture was applied, while they remained to a far extent unchanged when the non-ropy starter culture was used. It can be concluded from these studies that a preheating of milk at a temperature of 85°C (20 min) is optimal in regard to final yoghurt product characteristics. [source] Primary structure of a novel subunit in ba3 -cytochrome oxidase from thermus thermophilusPROTEIN SCIENCE, Issue 11 2000Tewfik Soulimane Abstract The ba3 -type cytochrome c oxidase from Thermus thermophilus is known as a two subunit enzyme. Deduced from the crystal structure of this enzyme, we discovered the presence of an additional transmembrane helix "subunit IIa" spanning the membrane. The hydrophobic N-terminally blocked protein was isolated in high yield using high-performance liquid chromatography. Its complete amino acid sequence was determined by a combination of automated Edman degradation of both the deformylated and the cyanogen bromide cleaved protein and automated C-terminal sequencing of the native protein. The molecular mass of 3,794 Da as determined by MALDI-MS and by ESI requires the N-terminal methionine to be formylated and is in good agreement with the value calculated from the formylmethionine containing sequence (3,766.5 Da + 28 Da = 3,794.5 Da). This subunit consits of 34 residues forming one helix across the membrane (Lys5-Ala34), which corresponds in space to the first transmembrane helix of subunit II of the cytochrome c oxidases from Paracoccus denitrificans and bovine heart, however, with opposite polarity. It is 35% identical to subunit IV of the ba3 -cytochrome oxidase from Natronobacterium pharaonis. The open reading frame encoding this new subunit IIa (cbaD) is located upstream of cbaB in the same operon as the genes for subunit I (cbaA) and subunit II (cbaB). [source] A study of Streptococcus thermophilus proteome by integrated analytical procedures and differential expression investigationsPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 1 2006Simona Arena Abstract Streptococcus thermophilus is a Gram-positive bacterium belonging to the group of lactic acid bacteria, among which several genera play an essential role in manufacture of food products. Recently, a genomic consortium sequenced and annotated its entire genome, which has been demonstrated to contain 1900 coding sequences. In this study, we have revealed the expression products of almost 200 different genes using a proteomic strategy combining 2-DE plus MALDI-TOF PMF and differential 1-DE plus ,LC-ESI-IT-MS/MS. Thus, a number of cellular pathways related to important physiological processes were described at the proteomic level. Almost 50 genes were related to multiple electrophoretic species, whose heterogeneity was mainly due to variability in pI values. A 2-DE reference map obtained for lactose-grown cells was compared with those obtained after heat, cold, acid, oxidative and starvation stresses. Protein up/down-regulation measurements demonstrated that adaptation to different environmental challenges may involve the contribution of unique as well as combined physiological mechanisms. Common regulatory sites in the promoter region of genes whose expression was induced after stress were identified. These results provide a better comprehension of biochemical processes related to stress resistance in S. thermophilus, allowing defining the molecular bases of adaptative responses or markers for the identification of strains with potential industrial applications. [source] Identification of a potent immunostimulatory oligodeoxynucleotide from Streptococcus thermophilus lacZANIMAL SCIENCE JOURNAL, Issue 5 2009Takeshi SHIMOSATO ABSTRACT Immunostimulatory sequences of oligodeoxynucleotides (ODNs), such as CpG ODNs, are potent stimulators of innate immunity. Here, we identified a strong immunostimulatory CpG ODN, which we named MsST, from the lac Z gene of Streptococcus (S.) thermophilus ATCC19258, and we evaluated its immune functions. In in vitro studies, MsST had a similar ability as the murine prototype CpG ODN 1555 to induce inflammatory cytokine production and cell proliferation. In mouse splenocytes, MsST increased the number of CD80+CD11c+and CD86+CD11c+ dendritic cells and CD4+CD25+ regulatory T cells. We also analyzed the effects of MsST on the expression of regulatory cytokines by real-time quantitative PCR. MsST was more potent at inducing interleukin-10 expression than the ODN control 1612, indicating that MsST can augment the regulatory T cell response via Toll-like receptor 9, which plays an important role in suppressing T helper type 2 responses. These results suggest that S. thermophilus, whose genes include a strong Immunostimulatory sequence-ODN, is a good candidate for a starter culture to develop new physiologically functional foods and feeds. [source] Major microbiota of lactic acid bacteria from Matsoni, a traditional Georgian fermented milkANIMAL SCIENCE JOURNAL, Issue 1 2007Kenji UCHIDA ABSTRACT A total of 26 samples of Matsoni were collected in Georgia. From these samples 80 strains of lactic acid cocci and 173 strains of lactobacilli were isolated. The number of lactic acid bacteria varied between 105 and 1010 colony forming unit (CFU)/mL. All the isolated lactic acid bacteria were thermophilic bacteria that could grow at 45°C. The predominant lactic acid bacteria were Streptococcus thermophilus and Lactobacillus delbrueckii spp. bulgaricus in 25 samples of Matsoni, while Lactobacillus helveticus was also a predominant species, together with the two previous species in one sample of Matsoni. We showed there was diversity in both S. thermophilus and L. delbrueckii spp. bulgaricus at the strain level by randomly amplified polymorphic DNA analysis. [source] Structure of the putative thioesterase protein TTHA1846 from Thermus thermophilus HB8 complexed with coenzyme A and a zinc ionACTA CRYSTALLOGRAPHICA SECTION D, Issue 8 2009Toshiaki Hosaka TTHA1846 is a conserved hypothetical protein from Thermus thermophilus HB8 with a molecular mass of 15.1,kDa that belongs to the thioesterase superfamily (Pfam 03061). Here, the 1.9,Å resolution crystal structure of TTHA1846 from T. thermophilus is reported. The crystal structure is a dimer of dimers. Each subunit adopts the so-called hot-dog fold composed of five antiparallel ,-strands flanked on one side by a rather long ,-helix and shares structural similarity to a number of thioesterases. Unexpectedly, TTHA1846 binds one metal ion and one ligand per subunit. The ligand density was modelled as coenzyme A (CoA). Its structure was confirmed by MALDI,TOF mass spectrometry and electron-density mapping. X-ray absorption fine-structure (XAFS) measurement of the crystal unambiguously characterized the metal ion as zinc. The zinc ion is tetrahedrally coordinated by the side chains of Asp18, His22 and Glu50 and the CoA thiol group. This is the first structural report of the interaction of CoA with a zinc ion. From structural and database analyses, it was speculated that the zinc ion may play an inhibitory role in the enzymatic activity. [source] Structure of the nondiscriminating aspartyl-tRNA synthetase from the crenarchaeon Sulfolobus tokodaii strain 7 reveals the recognition mechanism for two different tRNA anticodonsACTA CRYSTALLOGRAPHICA SECTION D, Issue 10 2007Yoshiteru Sato In protein synthesis, 20 types of aminoacyl-tRNA synthetase (aaRS) are generally required in order to distinguish between the 20 types of amino acid so that each achieves strict recognition of the cognate amino acid and the cognate tRNA. In the crenarchaeon Sulfolobus tokodaii strain 7 (St), however, asparaginyl-tRNA synthetase (AsnRS) is missing. It is believed that AspRS instead produces Asp-tRNAAsn in addition to Asp-tRNAAsp. In order to reveal the recognition mechanism for the two anticodons, GUC for aspartate and GUU for asparagine, the crystal structure of St -AspRS (nondiscriminating type) has been determined at 2.3,Å resolution as the first example of the nondiscriminating type of AspRS from crenarchaea. A structural comparison with structures of discriminating AspRSs indicates that the structures are similar to each other overall and that the catalytic domain is highly conserved as expected. In the N-terminal domain, however, the binding site for the third anticodon nucleotide is modified to accept two pyrimidine bases, C and U, but not purine bases. The C base can bind to form a hydrogen bond to the surrounding main-chain amide group in the discriminating AspRS, while in the nondiscriminating AspRS the corresponding amino-acid residue is replaced by proline, which has no amide H atom for hydrogen-bond formation, thus allowing the U base to be accommodated in this site. In addition, the residues that cover the base plane are missing in the nondiscriminating AspRS. These amino-acid changes make it possible for both C and U to be accepted by the nondiscriminating AspRS. It is speculated that this type of nondiscriminating AspRS has been introduced into Thermus thermophilus through horizontal gene transfer. [source] | |||||||||||||||||||||||||||||||