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[crystal structure of thermitase and stability of subtilisins].crystal structure of thermitase, a serine proteinase from thermoactinomyces vulgaris, has been determined by x-ray diffraction at 1.4 a resolution. the atomic model of thermitase refined to an r-factor of 0.149 contains 1997 protein atoms, 182 water molecules and 2 ca2+ ions. the tertiary structure of thermitase is similar to that of subtilisin bpn'. the greatest variations are connected with insertions and deletions in the amino acid sequence, which are located on the surface of the molecule. h ...19902198028
viable endospores of thermoactinomyces vulgaris in lake sediments as indicators of agricultural history.bacteria of the genus thermoactinomyces form endospores with an extreme longevity in natural habitats. we isolated thermoactinomyces sacchari from 9,000-year-old varved (annually laminated) sediment; thus, t. sacchari is probably one of the oldest known living organisms. more importantly, we tested and verified the hypothesis that there is a relationship between concentrations of dormant, viable endospores of t. vulgaris in lake sediments and the extent of agriculture in the catchments of the la ...19902202253
[study of the thrombolytic and fibrinolytic properties of thiol- dependent serine proteinase (tsp) from thermoactinomyces vulgaris in vivo].experiments on male albino rats showed that the thyol-dependent serine proteinase (tsp) dissolved the thrombus in the jugular vein for 2-5 hr. intravenous injection of tsp activated the fibrinolytic system of intact animals by increasing the levels of the plasminogen activator and plasmin in the euglobulin fraction. the response of the fibrinolytic system on the intravenous injection of tsp (2mg/200 g) was different: in some rats, fibrinolysis was activated, while on others, it was inhibited. ts ...19902281048
serum igg antibody concentrations against environmental microbes in mares and foals during different seasons and effect of stabling practices.over periods of 22 and 14 months, igg antibody concentrations in serum samples obtained monthly from 14 mares and 19 foals, respectively, were measured by use of elisa against antigens of the following environmental microbes: aspergillus umbrosus, penicillium brevicompactum, rhodotorula glutinis, absidia corymbifera, aspergillus fumigatus, humicola grisea, micropolyspora faeni, and thermoactinomyces vulgaris. the mares and foals were on pasture from early june until early october, then were stab ...19902327613
class-specific antibodies during follow up of patients with farmer's lung.sequential serum samples of 13 patients with acute farmer's lung (fl) taken during a follow-up of 18-36 months, were tested for antibodies of immunoglobulin g (igg), iga, igm and ige classes against thermoactinomyces vulgaris and micropolyspora faeni, and compared with contemporary lung function parameters. in the acute phase, antibodies of several ig classes were present, those of igg and iga being most common. at the end of the follow-up, the mean values of all antibody titres were lower than ...19902340884
type iii hypersensitivity reaction in mushroom growers.some respiratory symptoms in mushroom growers such as mushroom worker's lung develop by inhalation of certain agents arising from the environment of mushroom cultivation. recently we observed mushroom workers who had respiratory symptoms which might be type iii hypersensitivity reaction to the antigen of pleurotus floridae. we gave questionnaires to all the mushroom growers at one of the biggest cultivation areas of mushrooms, pleurotus floridae in pocheon, kyunggi province. those with respirato ...19911742253
[a study on interleukin 2 and soluble interleukin 2 receptor in serum and bronchoalveolar lavage fluid from patients with farmer's lung].interleukin 2 (il-2) and soluble interleukin 2 receptor (il-2r) were assayed in serum and bronchoalveolar lavage fluid (balf) from patients with farmer's lung. patients with farmer's lung had significantly higher levels of soluble il-2r in serum and balf than normal controls. the level of soluble il-2r was correlated with the disease activity, and fell to within the normal range after successful treatment with corticosteroids. on the other hand, asymptomatic dairy farmers with precipitating anti ...19911770680
thermophilic actinomycetes associated with agro-environment of punjab state (india).thermophilic actinomycetes (tas) are unique high temperature aerobic bacteria which belong to the group actinomycetes. while working on hypersensitivity pneumonitis (hp), an immunological disorder, resulting from the inhalation of spores of thermophilic actinomycetes, it was considered worthwhile to record their occurrence in the environment of punjab. thermophilic actinomycetes isolated by spread plate method from nine different substrates of the punjab environment were identified as 8 differen ...19911813626
studies on properties of thermostable beta-amylase from thermoactinomyces sp.v4.a thermostable beta-amylase from thermoactinomyces sp. v4 isolated from soil is reported. the enzyme has optimum activity at 70 degrees c and ph6. it is stable at 50 degrees c (4h) and retains 92% of activity at 55 degrees c (2h). the extent of hydrolysis soluble starch is about 77% and the product is identified as beta-maltose by paper chromatography and rotatory examination. sulfhydryl reagent has no effect on the beta-amylase activity.19911839610
occurrence and significance of precipitating antibodies against thermophilic actinomycetes in the sera of dairy herd workers, nangali, delhi.the study was prompted by the lack of information on the role of thermophilic actinomycetes in hypersensitivity pneumonitis in india. it reports the occurrence of precipitating antibodies against clinically important thermophilic actinomycetes in the sera of a population sample of dairy herd workers, nangali, delhi. of 112 workers investigated, 28 (25%) showed precipitins against faenia rectivirgula, 4 (3.2%) against saccharomonospora viridis, 2 against thermoactinomyces thalpophilus and one eac ...19911867472
thermostable phenylalanine dehydrogenase of thermoactinomyces intermedius: cloning, expression, and sequencing of its gene.the gene encoding the thermostable phenylalanine dehydrogenase [ec 1.4.1.-] of a thermophile, thermoactinomyces intermedius, was cloned and its complete dna sequence was determined. the phenylalanine dehydrogenase gene (pdh) consists of 1,098 nucleotides and encodes 366 amino acid residues corresponding to the subunit (mr 41,000) of the hexameric enzyme. the amino acid sequence deduced from the nucleotide sequence of the pdh gene of t. intermedius was 56.0 and 42.1% homologous to those of the ph ...19911880121
[immunologic response to microorganisms from a polluted ventilation system in a working environment].authors point out the morbidity of employees working in ventilation systems contaminated with various microorganisms. they analysed 96 workers exposed to air conditioning system (group a), and 71 workers (group b) breathing normal ambient air. the workers of both groups were subjected clinically by functionally and immunologically. preparation of antigens "mmm" (monday morning miseries) was used as an original method by ajello et al. for producing antigens from systemic mycotic agents and subseq ...19911916435
molecular cloning and primary structure of thermoactinomyces vulgaris carboxypeptidase t. a metalloenzyme endowed with dual substrate specificity.a gene coding for an extracellular zn-carboxypeptidase of thermoactinomyces vulgaris has been cloned and sequenced (embl x56901). this enzyme named carboxypeptidase t reveals simultaneously both types of substrate specificity characteristic of mammalian carboxypeptidases a and b. the carboxypeptidase t gene is primarily expressed in e. coli as a non-active preproenzyme with an additional 98 amino acid residues at the n-terminus. primary structure alignment of mature carboxypeptidase t and mammal ...19911936254
[a thermostable alpha-amylase from thermoactinomyces vulgaris: purification and characterization].alpha-amylase from thermoactinomyces vulgaris (strain 94-2a) was purified by cellulose chromatography and gel-chromatography on sephadex g-75 and subsequently characterised. the enzyme shows a single band in the polyacrylamide gel electrophoresis (page). the isoelectric point was determined to be ph 5.4, and the molecular mass was estimated as 53,000 dalton by page. the amino acid composition was determined; it shows characteristics of other microbial alpha-amylases. a comparison of the n-termin ...19911953690
circulating antibodies against thermophilic actinomycetes in farmers and mushroom workers.sera from individuals exposed to moldy hay, mushroom compost and unexposed individuals, were studied for antibodies against thermophilic actinomycetes and aspergillus fumigatus. antigens extracted from thermoactinomyces vulgaris strains isolated from china were used in the study along with antigens from standard strains to screen the sera of individuals exposed to farms and unexposed controls from both china and the united states. it was found that very little antibody response to a. fumigatus a ...19911960394
distribution, purification, and characterization of thermostable phenylalanine dehydrogenase from thermophilic actinomycetes.phenylalanine dehydrogenase (l-phenylalanine:nad oxidoreductase, deaminating; ec 1.4.1.-) was found in various thermophilic actinomycetes. we purified the enzyme to homogeneity from thermoactinomyces intermedius ifo 14230 by heat treatment and by red sepharose 4b, deae-toyopearl, sepharose cl-4b, and sephadex g-100 chromatographies with a 13% yield. the relative molecular weight of the native enzyme was estimated to be about 270,000 by gel filtration. the enzyme consists of six subunits identica ...19912061279
comparison of methods of production of farmer's lung antigens.the extracellular growth products of micropolyspora faeni, thermoactinomyces vulgaris and thermoactinomyces candidus were prepared by three methods; (1) double dialysis; (2) growth in tryptone soya broth (tsb) and (3) synthetic broth (aoac). it was found that double dialysis was superior to growth in tsb or aoac for the production of m. faeni antigens, as determined by immunoblot. double dialysis, tsb and aoac were similar for antigens of t. vulgaris and t. candidus. both organisms showed bands ...19921282842
purification and characterization of a thermostable alkaline protease from alkalophilic thermoactinomyces sp. hs682.protease secreted into the culture medium by alkalophilic thermoactinomyces sp. hs682 was purified to an electrophoretically homogeneous state through only two chromatographies using butyl-toyopearl 650m and sp-toyopearl 650s columns. the purified enzyme has an apparent relative molecular mass of 25,000 according to gel filtration on a sephadex g-75 column and sds-page and an isoelectric point above 11.0. its proteolytic activity was inhibited by active-site inhibitors of serine protease, dfp an ...19921368301
[exogenous allergic alveolitis: assessment of antigen-specific igg antibodies using a new chemiluminescence method].the demonstration of elevated levels of antigen-specific igg antibodies is of importance for the diagnostic workup of patients with a suspected hypersensitivity pneumonitis. we established a solid phase immunoassay for the determination of these antibodies based on chemiluminescence (clia). the clia demonstrated a low level of unspecific binding. the reproducibility of the clia was evaluated by calculating the intra- and inter-variation; the variation coefficient was 4% (intra-variation) resp. 1 ...19921372432
[serine proteinase from the archaebacterium halobacterium mediterranei--an analog of eubacterium subtilisin].a homogeneous serine proteinase was isolated from cultural filtrates of the extreme halophilic bacteria halobacterium mediterranei 1538 using affinity chromatography on bacitracin-sepharose, ultrafiltration and gel filtration on sephadex g-75, with a 48% yield and 260-fold purification. the enzyme was completely inactivated by specific inhibitors of serine proteinases, pmsf and dfp, as well as by hg2+ and pcmb. the enzyme activity was strongly dependent of nacl concentration, the enzyme being in ...19921391225
primary structure of carboxypeptidase t: delineation of functionally relevant features in zn-carboxypeptidase family.the primary structure of carboxypeptidase t--a zn-dependent extracellular enzyme of thermoactinomyces vulgaris--was determined from the cloned cpt gene nucleotide sequence and compared to zn-carboxypeptidases from various organisms. the compilation and analysis of multiple alignment accompanied by consideration of available tertiary structure data have shown that in the overall spatial structure and active site arrangement cpt is similar to other enzymes constituting the zn-carboxypeptidase fami ...19921449602
farmer's lung disease in north-western india--a preliminary report.a study of farmer's lung (fl) disease was carried out in 197 subjects engaged in farming and having respiratory complaints of varying duration. it revealed that 13.2% of the subjects had precipitating antibodies against thermophilic actinomycetes, with faenia rectivirgula (micropolyspora faeni) alone accounting for 85% of the positive reactions. precipitating antibodies against thermoactinomyces vulgaris and t. thalpophilus were observed only in 1.5% and 0.5% of the subjects, respectively. two s ...19921459662
[case report of farmer's lung disease in siblings in kagoshima prefecture].two cases of farmer's lung disease in siblings are reported. a 54-year-old male farmer, who had been engaged in stock work for 20 years, presented to our clinic for the second episode of fever, productive cough and shortness of breath. chest roentgenogram revealed diffuse micronodular pattern, and mild hypoxemia was recognized on arterial blood gas analysis. cytology obtained from balf showed lymphocytosis, with especially increased okt3, okt4 positive cells and okt4/8 ratio. the diagnosis was c ...19921464985
[changes in type ii alveolar epithelial cells in experimental allergic alveolitis].changes of type ii alveolar epithelial cells (type ii cells) were studied in experimental allergic alveolitis by using enzyme histochemical, electron microscopic and morphometric techniques. the results showed that the number of type ii cells increased obviously after exposed to thermoactinomyces vulgaris (tv). the peak was on the 15th day and then decreased gradually till to normal on the 180th day. type ii cells could be divided into two populations in tv-treated rats, namely, the large type i ...19921473211
hypersensitivity pneumonitis in athymic nude mice. additional evidence of t cell dependency.we previously demonstrated that c57 black/6 mice develop lung lesions similar to human hypersensitivity pneumonitis (hp) by repeated transnasal administration of thermoactinomyces vulgaris (tv) antigen. to elucidate the role of t cells in the development of this disease, tv antigen (90 micrograms/day) was transnasally administered to athymic nude c57 black/nu/nu (nu) mice and their littermates (+/nu) three times a week for 3 wk. the nude mice developed minimal lung lesions, whereas their thymus- ...19921489144
crystal structure of carboxypeptidase t from thermoactinomyces vulgaris.the crystal structure of carboxypeptidase t from thermoactinomyces vulgaris has been determined at 0.235-nm resolution by x-ray diffraction. carboxypeptidase t is a remote homologue of mammalian zn-carboxypeptidases. in spite of the low degree of amino acid sequence identity, the three-dimensional structure of carboxypeptidase t is very similar to that of pancreatic carboxypeptidases a and b. the core of the protein molecule is formed by an eight-stranded mixed beta sheet. the active site is loc ...19921521526
a serine proteinase of an archaebacterium, halobacterium mediterranei. a homologue of eubacterial subtilisins.a homogeneous serine proteinase secreted by the extreme halophilic bacterium halobacterium mediterranei 1538 was isolated by affinity chromatography on bacitracin-sepharose with a yield of 48% (260-fold purification). the enzyme reveals an optimum for pyroglutamyl-ala-ala-leu p-nitroanilide hydrolysis at ph 8.0-8.5 (km 0.14 mm; k(cat). 36.9 s-1). its activity increases linearly with nacl concentration over the range 2-5 m. the substrate specificity of the enzyme is comparable with that of secret ...19921637313
molecular cloning and sequencing of the gene for a halophilic alkaline serine protease (halolysin) from an unidentified halophilic archaea strain (172p1) and expression of the gene in haloferax volcanii.the gene of a halophilic alkaline serine protease, halolysin, from an unidentified halophilic archaea (archaebacterium) was cloned and its nucleotide sequence was determined. the deduced amino acid sequence showed that halolysin consists of 411 amino acids, with a molecular weight of 41,963. the highest homology was found with thermitase from thermoactinomyces vulgaris. halolysin has a long c-terminal extension of approximately 120 amino acids which has not been found in other extracellular subt ...19921732209
suprageneric classification of thermoactinomyces vulgaris by nucleotide sequencing of 5s ribosomal rna.the 5s rrna nucleotide sequence of thermoactinomyces vulgaris was determined and compared with published sequences of representative gram-positive bacteria. the primary and secondary structure of the sequence is of the type characteristic of gram-positive bacteria that have dna with a low proportion of guanine plus cytosine. it was evident from the phylogenetic trees that t. vulgaris has little in common with actinomycetes but is related to the genus bacillus, showing a moderately high relations ...19937688999
[thermoactinomyces vulgaris as a cause of extrinsic allergic alveolitis].thermoactinomycetes are microorganisms similar to the aerobic actinomycetes that grow at temperatures between 32 65 degrees c and are usually found in soil, hay and warm environment. they are the agent of extrinsic allergy alveolitis producing an interstitial process of immunologic, generated by the inhalation of the environmental dust that contain spores from these fungi. there is one case of a middle age man who works in a boiler of a foundry company of province de buenos aires in argentina. t ...19938160571
mushroom worker's lung: serologic reactions to thermophilic actinomycetes present in the air of compost tunnels.vast numbers of spores of the thermophilic actinomycetes excellospora flexuosa, thermomonospora alba, t. curvata and t. fusca were collected from the air in fermentation tunnels during the spawning of mushroom compost, i.e. over 10(9) cfu m-3 of air. five different genera of fungi, namely, aspergillus, aureobasidium, cladosporium, penicillium and scytalidium, were found at only 10(3) cfu m-3 of air. agaricus bisporus, used for spawning, was absent. sera of 10 mushroom growers affected by mushroo ...19938326995
responses of horses affected with chronic obstructive pulmonary disease to inhalation challenges with mould antigens.eight control and 8 asymptomatic copd-affected horses were given, on separate occasions, inhalation challenges with extracts of micropolyspora faeni, aspergillus fumigatus and thermoactinomyces vulgaris. all horses were also given nebulised phosphate-buffered saline (pbs) challenges and 'natural challenges' (ncs), i.e. exposure to hay and straw, as control challenges. responses were assessed by clinical, pulmonary mechanics, arterial blood gas tensions, arterial blood ph and bronchoalveolar lava ...19938354208
evaluation of intradermal mould antigen testing in the diagnosis of equine chronic obstructive pulmonary disease.intradermal end-point titres for commercial aqueous extracts of micropolyspora faeni, thermoactinomyces vulgaris and aspergillus fumigatus were determined in control and chronic obstructive pulmonary disease (copd) affected horses. the intradermal end-point titres of control and copd-affected horses were not significantly different and values for individual horses for m. faeni, a. fumigatus and t. vulgaris were not correlated with the pulmonary dysfunction or with the bronchoalveolar lavage flui ...19938354210
site-directed mutagenesis of a hexapeptide segment involved in substrate recognition of phenylalanine dehydrogenase from thermoactinomyces intermedius.phenylalanine dehydrogenase from thermoactinomyces intermedius and leucine dehydrogenase from bacillus stearothermophilus show a 59% sequence similarity in their substrate-binding domains, although their substrate specificities are different. we prepared a phenylalanine dehydrogenase mutant enzyme whose inherent hexapeptide segment (124f-v-h-a-a-129r) in the substrate-binding domain was replaced by the corresponding part of leucine dehydrogenase (m-d-i-i-y-q) in order to investigate the mechanis ...19938407879
two bacteria causing farmer's lung: fine structure of thermoactinomyces vulgaris and saccharopolyspora rectivirgula.the fine structure of thermoactinomyces vulgaris and saccharopolyspora rectivirgula is described by transmission electron microscopy. these two bacteria are the most common microbes causing farmer's lung. the fine structure of hyphae, germination of endospores and the details of conidial wall layers of t. vulgaris, as well as the fine structure of septate hypha and globose, polygonal conidia of s. rectivirgula are described. the conidial wall of t. vulgaris consisted of an inner multilayered spo ...19938474530
the effect of cations on the thermophilic character of alkaline phosphatase from thermoactinomyces vulgaris.the effect of cations on the thermophilic character of alkaline phosphatase from thermoactinomyces vulgaris, is described. the optimal ph and temperature were 9.5 and 55 degrees c to 65 degrees c, respectively. the partial removal of cations with ethylene diamine tetraacetic acid converted the enzyme to mesophilic and susceptible to chemical denaturation. their complete removal caused complete inhibition. the addition of 0.3mm cobalt and 10mm magnesium added before heating were found to be optim ...19938490579
a neopullulanase-type alpha-amylase gene from thermoactinomyces vulgaris r-47.shimizu et al. and ourselves have reported some enzymatic properties of an alpha-amylase from thermoactinomyces vulgaris r-47 that hydrolyzed pullulan to produce panose [m. shimizu et al., agric. biol. chem., 42, 1681-1688 (1978); y. sakano et al., agric. biol. chem., 46, 1121-1129 (1982)]. in this study, we cloned a gene for an alpha-amylase, which was different from the one mentioned above but also hydrolyzed pullulan to produce panose, from t. vulgaris r-47, and analyzed the entire primary st ...19937763540
purification and properties of the highly thermostable alkaline protease from an alkaliphilic and thermophilic bacillus sp.thermostable alkaline protease from an alkaliphilic thermophile bacillus sp. b18' was purified by using deae- and cm-toyopearl 650m column chromatographies. molecular weights of the enzyme determined by sds-page and gel filtration were 30,000 and 28,000, respectively. the optimum ph and temperature toward the hydrolysis of casein were ph 12-13 and 85 degrees c, both of which are higher than those of a mesophilic alkaline protease from an alkaliphile, bacillus sp. b21-2. the enzyme was stable at ...19937764036
physiology of some actinomycete genera.actinomycetes are widespread in the environment and are mainly organotrophic. studies of their ecology have been primarily focussed on their detection and isolation, with comparatively little attention to the control mechanisms that determine their occurrence and behaviour in their natural environments. this session provided some diverse examples of approaches to this problem. several actinomycete genera produce motile spores. the significance of flagella proteins and factors influencing spore m ...19938140284
regulation of carboxypeptidase e. effect of ca2+ on enzyme activity and stability.carboxypeptidase e (cpe), an enzyme that functions in the post-translational processing of bioactive peptides, is a member of the metallocarboxypeptidase gene family. a 12-residue region of cpe has 70% amino acid identity with the bacterial enzyme carboxypeptidase t (cpt); in cpt, this region has been identified previously as the ca(2+)-binding region (teplyakov, a., polyakov, k., obmolova, g., strokopytov, b., kuranova, i., osterman, a., grishin, n., smulevitch, s., zagnitko, o., galperina, o., ...19948157647
identification of active site lysyl residues of phenylalanine dehydrogenase by chemical modification with methyl acetyl phosphate combined with site-directed mutagenesis.a monoanionic acetylation reagent, methyl acetyl phosphate, was used to acetylate lysyl residues of the recombinant thermostable phenylalanine dehydrogenase from thermoactinomyces intermedius. the enzyme was irreversibly inactivated with the reagent in a time- and dose-dependent manner. simultaneous addition of substrate and coenzyme markedly protected the enzyme from inactivation. acetylated lysyl residues presumably occurring at the active site were determined by differential modification; the ...19947706231
alpha-amylases from thermoactinomyces vulgaris: characteristics, primary structure and structure prediction.two amylolytic active protein fractions (named alpha-amylase 1 and alpha-amylase 2) were isolated from the bacterium thermoactinomyces vulgaris strain 94-2a. alpha-amylase 1 had a molecular mass of 51.6 kda, whereas alpha-amylase 2 consists of two fragments which have molecular masses of 17.0 and 34.6 kda, respectively. these two fragments are products from a proteolytic cleavage of alpha-amylase 1 at amino acid position 303 (tryptophan) by a serine protease (thermitase) which is also produced b ...19947822101
construction and characterization of chimeric enzyme consisting of an amino-terminal domain of phenylalanine dehydrogenase and a carboxy-terminal domain of leucine dehydrogenase.phenylalanine dehydrogenase of thermoactinomyces intermedius acts preferentially on l-phenylalanine and l-tyrosine, whereas leucine dehydrogenase of bacillus stearothermophilus acts almost exclusively on l-leucine and some other branched-chain l-amino acids. the two enzymes share a sequence similarity (47%). aiming at elucidation of the mechanism of substrate recognition by the two amino acid dehydrogenases, we have genetically constructed a chimeric enzyme consisting of an n-terminal domain of ...19947883771
phylogenetic diversity in the genus bacillus and comparative ribosomal protein at-l30 analyses of the genus thermoactinomyces and relatives.the ribosomal l30 proteins from strains of 27 species belonging to the genera bacillus, escherichia, staphylococcus, lactobacillus, leuconostoc and thermoactinomyces were analysed, together with at-l30 proteins from selected actinomycetes. the results of partial amino acid sequencing of l30 preparations revealed that the members of the genera escherichia, staphylococcus and thermoactinomyces were homogeneous within each genus. in contrast, phylogenetic diversity existed in the genus bacillus, wh ...19947921265
the gene amye(tv1) codes for a nonglucogenic alpha-amylase from thermoactinomyces vulgaris 94-2a in bacillus subtilis.we isolated the gene amye(tv1) from thermoactinomyces vulgaris 94-2a encoding a nonglucogenic alpha-amylase (amytv1). a chromosomal dna fragment of 2,247 bp contained an open reading frame of 483 codons, which was expressed in escherichia coli and bacillus subtilis. the deduced amino acid sequence of the amytv1 protein was confirmed by sequencing of several peptides derived from the enzyme isolated from a t. vulgaris 94-2a culture. the amino acid sequence was aligned with several known alpha-amy ...19947944369
cloning and sequencing of a serine proteinase gene from a thermophilic bacillus species and its expression in escherichia coli.the gene for a serine proteinase from a thermophilic bacillus species was identified by pcr amplification, and the complete gene was cloned after identification and isolation of suitably sized restriction fragments from southern blots by using the pcr product as a probe. two additional, distinct pcr products, which were shown to have been derived from other serine proteinase genes present in the thermophilic bacillus species, were also obtained. sequence analysis showed an open reading frame of ...19947993087
the purification, characterization, cloning and sequencing of the gene for a halostable and thermostable leucine dehydrogenase from thermoactinomyces intermedius.leucine dehydrogenase has been purified to homogeneity from a moderate thermophilic actinomycete, thermoactinomyces intermedius ifo 14230. the enzyme can be stored without loss of its activity at a low temperature (e.g., 4 degrees c) for over two years. the enzyme was more thermostable at higher concentrations of salts such as nacl and kcl. it retained about 90% of activity on incubation at 70 degrees c for at least 40 min in the presence of 3 m nacl. the michaelis constants for nad, l-leucine, ...19948020469
occurrence of thermophilic actinomycetes in natural substrates in nigeria.thermophilic actinomycetes were isolated from 163 (48.95%) of 333 samples of vegetable substrates and soil from different sites in anambra and enugu states in nigeria. thermoactinomyces (tha.) vulgaris was the most common, occurring in 32.4% of samples while tha. thalpophilus was isolated from 20.1%. tha. sacchari, saccharomonospora (sam.) viridis and saccharopolyspora (sap.) rectivirgula were isolated from 3-10.5% of the samples examined. streptomyces (stm.) thermovulgaris occurred in 2.7% and ...19948060118
a convenient enzymatic synthesis of 4(2)-alpha-isomaltosylisomaltose using thermoactinomyces vulgaris r-47 alpha-amylase ii (tva ii). 19948087809
establishment of oil-degrading bacteria associated with cyanobacteria in oil-polluted soil.a unique natural microbial cocktail with promising potential for remediating oil-polluted desert in the gulf region is reported. oil-degrading micro-organisms immobilized within dense cyanobacterial mats on oily coasts of the arabian gulf were successfully established in oil-contaminated sand. those micro-organisms biodegraded 50% of the oil within 10-20 weeks. nocardioforms belonging to the genus rhodococcus predominated in the first few weeks, but after 22 weeks pseudomonas spp. increased, sha ...19957698954
comparison of primary structures and substrate specificities of two pullulan-hydrolyzing alpha-amylases, tva i and tva ii, from thermoactinomyces vulgaris r-47.thermoactinomyces vulgaris r-47 produces two alpha-amylases, tva i, an extracellular enzyme, and tva ii, an intracellular enzyme. both enzymes hydrolyze pullulan to produce panose, and also hydrolyze cyclodextrins. we cloned and sequenced the tva i gene. the tva i gene consisted of 1833 base pairs, and the deduced primary structure was composed of 611 amino-acid residues, including an n-terminal signal sequence consisting of 29 amino-acid residues. the similarity between the amino-acid sequence ...19957548164
chemotaxis of alveolar macrophages and neutrophils in response to microbial products derived from organic dust.the inhalation of organic dust has been implicated to cause a large number of occupational lung diseases. a common event in these diseases is an inflammatory reaction affecting airway and alveolar tissue that is characterized by the recruitment of different types of inflammatory cells. mechanisms of chemotaxis of alveolar macrophages (ams) and neutrophils (pmns) in response to microbial products derived from organic dust were studied. seven agents known to be etiological agents causing respirato ...19957588495
carboxypeptidase t. 19957674954
dairy farmers have increased methacholine bronchial responsiveness independent of sensitization to mold antigens.patients with farmer's lung disease (fld) and dairy farmers have nonspecific bronchial hyperresponsiveness. to examine the factors determining bronchial hyperresponsiveness among dairy farmers, we studied airway functions, airway responses to inhaled methacholine, serum total ige levels, and antigen-specific ige levels in 37 dairy farmers and 11 local control subjects. the 37 dairy farmers consisted of three groups; 12 farmers with episodes of fld (fld group), 13 farmers with serum antibody to m ...19957767522
chemotaxis of alveolar macrophages and neutrophils in response to microbial products derived from organic dust.mechanisms of chemotaxis of alveolar macrophages (ams) and neutrophils (pmns) in response to microbial products derived from organic dust were studied using the blindwell chemotaxis chamber technique. seven different known etiological agents causing respiratory symptoms were used for experiments: cell extract and endotoxin from pantoea agglomerans (synonyms: erwinia herbicola, enterobacter agglomerans), cell extracts from thermoactinomyces vulgaris and aspergillus fumigatus, protease from bacill ...19958705013
thermitase - kinetic differentiation to the subtilisins.in the last 12 years more than 80 peptidyl substrates and substrate analogous inhibitors were synthesized in our group and used to characterize thermitase, a serine proteinase of the subtilisin family produced by thermoactinomyces vulgaris. the kinetic parameters of hydrolysis and inhibition reactions, respectively, with thermitase were determined and compared with those found with related enzymes. conclusions with respect to the extent of the active site of thermitase and to the specificity of ...19968796317
the medical and epidemiologic effects on workers of the levels of airborne thermoactinomyces spp. spores present in australian raw sugar mills.the objectives of the study were to determine whether there was a significant risk of members of the work force of raw sugar mills developing bagassosis. airborne thermoactinomyces sacchari spores were measured to determine whether they were sufficient to cause acute bagassosis, and whether there was any evidence of previous exposure to sufficient airborne t. sacchari spores to cause the development of chronic bagassosis in any of the work force. monitoring of total airborne bacteria spore conce ...19968931309
hypersensitivity pneumonitis in workers exposed to esparto grass (stipa tenacissima) fibers.esparto grass (stipa tenacissima), which is commonly found in the mediterranean countries, has a wide variety of uses. five stucco makers who had cough, dyspnea, malaise, and fever after exposure to esparto fiber used in their jobs showed a significant decrease in symptoms when they were away from work. precipitating antibodies against an esparto extract were found in the sera of all patients. specific igg antibodies against the esparto extract were also demonstrated in all patient sera, as were ...19968939163
microflora and acarofauna of bed dust from homes in upper silesia, poland.pyroglyphid mites are considered a major cause of house dust allergy. the occurrence and possible pathogenic role of other biologic components of house dust, in particular bacteria, has received less attention.19968564627
purification and characterization of a thermostable alkaline protease from thermoactinomyces sp. e79 and the dna sequence of the encoding gene.a thermophilic thermoactinomyces sp. e79 producing a highly thermostable alkaline protease was isolated from soil. the protease, produced extracellularly by thermoactinomyces sp. e79, was purified by deae-sepharose cl-6b and butyl-toyopearl 650m column chromatography. the relative molecular mass was estimated to be 31,000 by sds-polyacrylamide gel electrophoresis. enzyme activity was inhibited by phenylmethylsulfonyl fluoride, suggesting the enzyme to be a serine protease. the optimum temperatur ...19968704314
[the influence of organic dust on the chemotaxis of lung cells. experimental studies].mechanisms of chemotaxis of alveolar macrophages (ams) and neutrophils (pmns) in response to microbial products derived from organic dust were studied using blindwell chemotaxis chamber technique. seven different agents (extract and endotoxin of erwinia herbicola, extracts from thermoactinomyces vulgaris and aspergillus fumigatus, thermophilic protease and two preparations of glucans), were used for experiments. these agents were evaluated for their ability to direct attraction of ams and pmns a ...19969190243
[indoor air studies of mould fungus contamination of homes of selected patients with bronchial asthma (with special regard to evaluation problems)].investigations of indoor air of the homes of seven patients with asthma bronchiale who showed up with positive reactions following intracutaneous application of fungal allergens revealed that their places of residence were contaminated by fungal and bacterial spores. the number of colony forming units of mesophilic fungal spores of the indoor air ranged from 100 to 1000 cfu/m3 and this was much higher than the mould flora of the outdoor air determined simultaneously. the major fungi species foun ...19969376049
construction of a new leucine dehydrogenase with preferred specificity for nadp+ by site-directed mutagenesis of the strictly nad+-specific enzyme.on the basis of sequence comparison between nad+-dependent leucine dehydrogenase (leudh) from thermoactinomyces intermedius and nadp+-dependent dehydrogenases, a set of amino acid residues that are supposed to determine the coenzyme specificity of leudh were assigned. systematic replacement of these amino acids by others was done with the aim to switch its natural coenzyme specificity to a new one preferring nadp+. single d203a, double d203a-i204r and triple d203a-i204r-d210r mutation enzymes we ...19979278282
isolation and growth physiology of novel thermoactinomycetes.a total of 34 thermophilic isolates identified as members of the genus thermoactinomyces by a range of chemotaxonomic, microscopic and determinative biochemical tests, were isolated from two acid soils. growth studies in shake flask and fermenter identified the isolates to be moderately acidophilic with growth occurring between ph 4.5 and 6.0 with optima at ph 5.0. the isolates differed considerably from known thermoactinomyces cultures in their ph profile, colony morphology and in several bioch ...19979449806
cloning and expression of an intracellular alkaline protease gene from alkalophilic thermoactinomyces sp. hs682.an intracellular alkaline serine protease gene of alkalophilic thermoactinomyces sp. hs682 was cloned and expressed in escherichia coli. sequence analysis showed a putative promoter region, a putative transcriptional termination signal, and an open reading frame of 963 bases, coding for a polypeptide of 321 amino acids. the protease expressed in e. coli was purified by deae-toyopearl 650m and sephadex g-75 chromatography. the n-terminal sequence (30 amino acids) of the purified protein was coinc ...19979058969
purification and characterization of serine proteinase of the glu,asp-specific enzyme family from thermoactinomyces species.enzyme catalyzing hydrolysis of a substrate of glu,asp-specific proteinases (z-glu-pna) and cleaving bond glu13-ala14 in the oxidized insulin b chain was purified to homogeneity from the culture medium of thermoactinomyces species using hydrophobic chromatography on phenyl-sepharose cl 4b as the key purification step. the molecular weight of the proteinase is 23 kd. the enzyme is completely inhibited by diisopropyl fluorophosphate and is stable at ph 5-11. the ph optimum for the hydrolysis of az ...19979159871
cloning, sequencing and overexpression of the leucine dehydrogenase gene from bacillus cereus.the l-leucine dehydrogenase gene from bacillus cereus (dsm 626) was cloned from a partial genomic library and sequenced. the open reading frame has 1101 bp and codes for a protein of 39.9 kda. the deduced amino acid sequence of the leudh from b. cereus shares 70-80% identity with leudh's from the thermophilic strains b. stearothermophilus and thermoactinomyces intermedius. the active protein was overexpressed in escherichia coli to yield approximately 30% of the total soluble protein.19979188201
expression of the carboxypeptidase t gene from thermoactinomyces vulgaris in stable protoplast type l-forms of proteus mirabilis.the structural gene of the carboxypeptidase t (cpt) was successfully expressed in cell wall-less l-form cells of proteus mirabilis. the dna sequence encoding the phoa leader peptide was fused with a truncated cpt gene encoding the mature enzyme. the modified gene in a puc-based kanamycin resistance vector under the control of the lac promoter was transformed into l-form cells of p. mirabilis. they were able to produce the recombinant cpt both as a secretory and as a cell-bound insoluble form. th ...19989503606
a cysteine-dependent serine protease associated with the dormant spores of bacillus cereus: purification of the protein and cloning of the corresponding gene.subtilisin-like serine protease, which is associated with the dormant spores of bacillus cereus, was solubilized by washing the spores with 2 m kcl and purified to homogeneity by carbobenzoxy-d-phenylalanine-liganded affinity column chromatography and hydrophobic interaction column chromatography. enzyme activity was completely inhibited by reagents for sulfhydryl groups such as hgcl2 as well as by conventional subtilisin inhibitors, suggesting the enzyme to be cysteine-dependent. the enzyme ret ...19989532782
conversion of neopullulanase-alpha-amylase from thermoactinomyces vulgaris r-47 into an amylopullulanse-type enzyme.tva i, an alpha-amylase from thermoactinomyces vulgaris r-47, is a versatile enzyme which hydrolyzes the alpha-(1-->4)-glucosidic linkages of pullulan to produce panose, known as neopullulanase activity, and the alpha-(1-->6)-glucosidic linkages of certain oligosaccharides. we modified the ala-357, gln-359, and tyr-360 residues located in region ii, one of the four regions conserved in alpha-amylase family enzymes, and deleted 11 consecutive amino acid residues located after the c-terminus of re ...19989538203
characteristics of airborne actinomycete spores.airborne actinomycete spores, important contaminants in occupational and residential environments, were studied with respect to their (i) release into the air, (ii) aerodynamic and physical size while airborne, and (iii) survival after collection onto agar with an impactor. three actinomycete species were selected for the tests to exemplify the three main spore types: streptomyces albus for arthrospores, micromonospora halophytica for aleuriospores, and thermoactinomyces vulgaris for endospores. ...19989758803
analysis of airborne actinomycete spores with fluorogenic substrates.the reactions between seven fluorogenic substrates and different groups of enzymes, esterases, lipases, phosphatases, and dehydrogenases, were studied in a search for a new method for the detection of actinomycete spores. fluorescence measurement was chosen as a fast and sensitive method for microbial analysis. the focus of the research was on the spores of important air contaminants: streptomyces albus and thermoactinomyces vulgaris. for the measurement of the enzymatic activity, the chosen flu ...19989797299
effect of inhalation of organic dust-derived microbial agents on the pulmonary phagocytic oxidative metabolism of guinea pigs.the effect of inhalation exposure of various biological agents associated with organic dusts on the function of guinea pigs pulmonary phagocytes was investigated. agents included antigens of erwinia herbicola, thermoactinomyces vulgaris, and aspergillus fumigatus; endotoxin of erwinia herbicola; bacterial protease; and a fungal glucan preparation. pulmonary parameters monitored in this study were cellular differential counts from bronchoalveolar lavage, and superoxide anion and/or hydrogen perox ...19989447225
novel rhodococci and other mycolate actinomycetes from the deep sea.a large number of mycolate actinomycetes have been recovered from deep-sea sediments in the nw pacific ocean using selective isolation methods. the isolates were putatively assigned to the genus rhodococcus on the basis of colony characteristics and mycolic acid profiles. the diversity among these isolates and their relationship to type strains of rhodococcus and other mycolate taxa were assessed by curie point pyrolysis mass spectrometry (pyms). three major (a, c, d) and two minor (b, e) groups ...199810068786
[cloning and expression of a serine proteinase gene from thermoactinomyces sp. in bacillus subtilis cells].a gene coding for thermostable serine protease from thermoactinomyces sp. k50 is cloned and expressed in bacillus subtilis cells. restriction map of cloned dna fragment is determined. thermostability and temperature optimum of proteolytic activity of the cloned gene product are lower than those of the natural proteinase of thermoactinomyces sp. k50. serine protease, a product of cloned gene, is highly sensitive to proteolysis and its degradation can be prevented by ca2+ ions.199910190105
crystal structure of thermoactinomyces vulgaris r-47 alpha-amylase ii (tvaii) hydrolyzing cyclodextrins and pullulan at 2.6 a resolution.the crystal structure of thermoactinomyces vulgaris r-47 alpha-amylase ii (tvaii) has been determined by multiple isomorphous replacement at 2.6 a resolution. tvaii was crystallized in an orthorhombic system with the space group p212121 and the cell dimensions a=118.5 a, b=119.5 a, c=114.5 a. there are two molecules in an asymmetric unit, related by the non-crystallographic 2-fold symmetry. diffraction data were collected at 113 k and the cell dimensions reduced to a=114.6 a, b=117.9 a, c=114.2 ...199910222200
isolation and partial characterisation of extracellular keratinase from a wool degrading thermophilic actinomycete strain thermoactinomyces candidus.the keratinase production by the thermophilic actinomycete strain thermoactinomyces candidus was induced by sheep wool as the sole source of carbon and nitrogen in the cultivation medium. for complete digestion of wool by the above strain, both keratinolytic serine proteinase and cellular reduction of disulfide bonds were involved. evidence was presented that substrate induction was a major regulatory mechanism and the keratinase biosynthesis was not completely repressed by addition of other car ...199910408094
igg antibodies, chronic bronchitis, and pulmonary function values in farmer's lung patients and matched controls.we measured igg antibody levels against eight different microbes in farmer's lung (fl) patients an average of 14 years after the first diagnosed episode of fl and in matched controls. the study population consisted of 87 fl patients and 81 control farmers, matched by age, sex, and smoking habits. clinical studies included the measurement of igg antibody levels against absidia corymbifera, aspergillus umbrosus, a. fumigatus, humicola grisea, saccharopolyspora rectivirgula, penicillium brevicompac ...199910604554
directed evolution converts subtilisin e into a functional equivalent of thermitase.we used directed evolution to convert bacillus subtilis subtilisin e into an enzyme functionally equivalent to its thermophilic homolog thermitase from thermoactinomyces vulgaris. five generations of random mutagenesis, recombination and screening created subtilisin e 5-3h5, whose half-life at 83 degrees c (3.5 min) and temperature optimum for activity (topt, 76 degrees c) are identical with those of thermitase. the topt of the evolved enzyme is 17 degrees c higher and its half-life at 65 degree ...199910065710
transfer of 'thermoactinomyces glaucus' ifo 12530 and 'thermoactinomyces monosporus' ifo 14050 to the genus saccharomonospora as members of saccharomonospora glauca.two available strains of 'thermoactinomyces glaucus' and 'thermoactinomyces monosporus', 't. glaucus' ifo 12530 and 't. monosporus' ifo 14050, were considered not to be members of the genus thermoactinomyces and that they belonged to the genus saccharomonospora on the basis of the colors of colonies and 16s rdna sequences. some chemotaxonomic characteristics also showed that the two strains belong to the genus saccharomonospora. the two strains contained meso-diaminopimelic acid, galactose, and ...200012483576
synthesis of allysine ethylene acetal using phenylalanine dehydrogenase from thermoactinomyces intermedius.allysine ethylene acetal [(s)-2-amino-5-(1,3-dioxolan-2-yl)-pentanoic acid (2)] was prepared from the corresponding keto acid by reductive amination using phenylalanine dehydrogenase (pdh) from thermoactinomyces intermedius atcc 33205. glutamate, alanine, and leucine dehydrogenases, and pdh from sporosarcina species (listed in order of increasing effectiveness) also gave the desired amino acid but were less effective. the reaction requires ammonia and nadh. nad produced during the reaction was r ...200010713207
structure, specificity and function of cyclomaltodextrinase, a multispecific enzyme of the alpha-amylase family.cyclomaltodextrinase (cdase, ec 3.2.1.54), maltogenic amylase (ec 3. 2.1.133), and neopullulanase (ec 3.2.1.135) are reported to be capable of hydrolyzing all or two of the following three types of substrates: cyclomaltodextrins (cds); pullulan; and starch. these enzymes hydrolyze cds and starch to maltose and pullulan to panose by cleavage of alpha-1,4 glycosidic bonds whereas alpha-amylases essentially lack activity on cds and pullulan. they also catalyze transglycosylation of oligosaccharides ...200010825529
phylogenetic analysis of the genus thermoactinomyces based on 16s rdna sequences.the almost complete 16s rdna sequences of the type strains of eight validly described species and two invalid species of the genus thermoactinomyces were determined and phylogenetically analysed. the validly described thermoactinomyces species formed phylogenetic lineages related to the family bacillaceae, as shown previously. however, the available strains of 'thermoactinomyces glaucus' and 'thermoactinomyces monosporus' exhibited their closest phylogenetic affinities not to the genus thermoact ...200010843048
production of cutinolytic esterase by filamentous bacteria.thirty-eight strains of filamentous bacteria, many of which are thermophilic or thermotolerant and commonly found in composts and mouldy fodders, were examined for their ability to produce cutinolytic esterase (cutinase) in culture media supplemented with cutin, suberin or cutin-containing agricultural by-products. initially, the ability of culture supernatants to hydrolyse the artificial substrate p-nitrophenyl butyrate was determined by spectrophotometric assays. only one bacterium, thermoacti ...200010886609
specificity of a neutral zn-dependent proteinase from thermoactinomyces sacchari toward the oxidized insulin b chain.the proteolytic specificity of the neutral zn-dependent proteinase from thermoactinomyces sacchari was determined by analysis of the peptides obtained after incubation with the oxidized insulin b chain as a substrate. the enzyme is an endopeptidase with broad specificity. in total, 12 peptide bonds in the b chain of insulin were hydrolyzed. the major requirement is that a hydrophobic residue such as leu, val, or phe should participate with the alpha-amino group in the bond to be cleaved. however ...200010919403
serum kl-6 concentrations in dairy farmers.serum kl-6 (krebs von den lungen-6) has been recognized to be a marker for the activity of diffuse interstitial lung diseases. the purpose of the study is to evaluate serum kl-6 measurement as a marker for farmer's lung disease (fld).200010936139
dna-dna relatedness among thermoactinomyces species: thermoactinomyces candidus as a synonym of thermoactinomyces vulgaris and thermoactinomyces thalpophilus as a synonym of thermoactinomyces sacchari.dna-dna relatedness of all validly described thermoactinomyces species was determined to infer the genetic relationships between them. the levels of dna-dna relatedness among the type strains of thermoactinomyces species ranged from 2-5 to 92.8%. based on dna relatedness data, the type strains of thermoactinomyces intermedius, thermoactinomyces putidus, thermoactinomyces dichotomicus and thermoactinomyces peptonophilus were considered to be distinct species of the genus thermoactinomyces. howeve ...200011034503
cloning and expression of the ccpa gene encoding catabolite control protein from thermoactinomyces sp. e79.the gene ccpa encoding the catabolite control protein a (ccpa) of thermoactinomyces sp. e79 has been cloned and characterized. nucleotide sequence analysis of the ccpa clone showed that the cloned fragment contained the full structural gene for a protein of 346 amino acids. the predicted amino acid sequence shows similarity to the transcriptional regulators of the lacl-galr family; a highly conserved helix-turn-helix motif, which might bind to dna, was identified through comparison with regulato ...200011129608
analysis of catalytic residues of thermoactinomyces vulgaris r-47 alpha-amylase ii (tva ii) by site-directed mutagenesis.to confirm that the catalytic residues (asp325, glu354, and asp421) are necessary for the hydrolysis of starch, pullulan, and cyclodextrins, we constructed tva ii mutated by site-directed mutagenesis. the mutated enzymes (d325n, e354q, and d421n) had markedly reduced levels of activity, less than 0.006% of the wild type, indicating that these three residues are the catalytic sites for these substrates. even e354d had reduced levels of activity, less than 0.05% of wild type. these four mutated en ...200011210138
studies on the hydrolyzing mechanism for cyclodextrins of thermoactinomyces vulgaris r-47 alpha-amylase 2 (tvaii). x-ray structure of the mutant e354a complexed with beta-cyclodextrin, and kinetic analyses on cyclodextrins.crystals of the mutant e354a of thermoactinomyces vulgaris r-47 alpha-amylase 2 (tvaii) complexed with beta-cyclodextrin were prepared by a soaking method, and the diffraction data were collected at 100 k, using synchrotron radiation (spring-8). the crystals belong to an orthorhombic system with the space group p2(1)2(1)2(1) and cell dimensions a = 111.1 a, b = 117.7 a, c = 113.3 a, which is almost isomorphous with crystals of the wild-type tvaii, and the structure was refined to an r-factor = 0 ...200111226882
[a new thermostable proteinase from thermoactinomyces sp.27a. cloning and gene expression].a library of thermoactinomyces sp. 27a, producer of thermostable proteases of different groups, has been created. gene coding for thermostable neutral proteinase was cloned and expressed in bac. subtilis cells. restriction map for cloned dna fragment was created and physicochemical parameters of recombinant proteinase were characterized. the thermostability and optimum of proteolytic activity of the enzyme was lower than in the natural thermoactinomyces sp. strain, which can be due to heterologo ...200111236740
the deletion of amino-terminal domain in thermoactinomyces vulgaris r-47 alpha-amylases: effects of domain n on activity, specificity, stability and dimerization.thermoactinomyces vulgaris r-47 alpha-amylases, tva i and tva ii, have a domain n, which is an extra structure in the family 13 enzymes. to investigate the roles of domain n in tvas, we constructed tvas-deltan mutants which are deleted in domain n, and y14,16,68a and y41,82,95a mutants of tva ii. tvas-deltan were unstable under alkaline conditions, and their thermal stabilities were 10 degrees c lower than that of wild-types. the specific activities of tvas-deltan for pullulan, starch, cyclodext ...200111302176
structures of thermoactinomyces vulgaris r-47 alpha-amylase ii complexed with substrate analogues.the structures of thermoactinomyces vulgaris r-47 alpha-amylase ii mutant (d325ntva ii) complexed with substrate analogues, methyl beta-cyclodextrin (m beta-cd) and maltohexaose (g6), were solved by x-ray diffraction at 3.2 a and 3.3 a resolution, respectively. in d325ntva ii-m beta-cd complex, the orientation and binding-position of beta-cd in tva ii were identical to those in cyclodextin glucanotransferase (cgtase). the active site residues were essentialy conserved, while there are no residue ...200111330677
thermostable, alkalophilic and cellulase free xylanase production by thermoactinomyces thalophilus subgroup c.thermoactinomyces thalophilus produced cellulase free extracellular endo-1,4-beta-xylanase (ec 3.2.1.8) at 50 degrees c and ph 8.5. maximum xylanase production was achieved in fermentation medium using birchwood xylan as substrate after 96 h of growth at 50 degrees c. other agricultural substrates such as wheat bran, wheat straw, sugarcane bagasse and cornstover produced less xylanase. the crude enzyme preparation from mutant t. thalophilus p2 grown under optimised fermentation conditions showed ...200111339942
phylogenetic diversity of thermophilic actinomycetes and thermoactinomyces spp. isolated from mushroom composts in korea based on 16s rrna gene sequence analysis.forty one strains isolated from 21 samples of various mushroom composts in korea were analyzed by 16s rrna gene sequencing to investigate the phylogenetic diversity of thermophilic actinomycetes. the 25 strains of thermophilic actinomycete isolates were related to the five genera, pseudonocardia, saccharomonospora, saccharopolyspora, streptomyces, and thermobifida, within the order actinomycetales, and 16 strains were classified into the genus thermoactinomyces within the family bacillaceae. mos ...200111506914
random amplified ribosomal dna restriction analysis for rapid identification of thermophilic actinomycete-like bacteria involved in hypersensitivity pneumonitis.hypersensitivity pneumonitis (hp) is a pulmonary disease characterised by inflammation that can be caused by, amongst other substances, a subset of 4 thermophilic mycelial bacteria: saccharopolyspora rectivirgula, saccharomonospora viridis, thermoactinomyces sacchari, and thermoactinomyces vulgaris. air sampling analyses in highly contaminated environments are often performed to evaluate exposure to these species which are difficult and fastidious to identify by conventional techniques. the aim ...200111518332
role of phe286 in the recognition mechanism of cyclomaltooligosaccharides (cyclodextrins) by thermoactinomyces vulgaris r-47 alpha-amylase 2 (tvaii). x-ray structures of the mutant tvaiis, f286a and f286y, and kinetic analyses of the phe286-replaced mutant tvaiis.phe286 located in the center of the active site of alpha-amylase 2 from thermoactinomyces vulgaris r-47 (tvaii) plays an important role in the substrate recognition for cyclomaltooligosaccharides (cyclodextrins). the x-ray structures of mutant tvaiis with the replacement of phe286 by ala (f286a) and tyr (f286y) were determined at 3.2 a resolution. their structures have no significant differences from that of the wild-type enzyme. the kinetic analyses of phe286-replaced variants showed that the v ...200111527532
novel glucoamylase-type enzymes from thermoactinomyces vulgaris and methanococcus jannaschii whose genes are found in the flanking region of the alpha-amylase genes.a region downstream of the gene for pullulan-hydrolyzing alpha-amylase, tva ii, of thermoactinomyces vulgaris r-47 was sequenced, and an open reading frame encoding an enzyme homologous to glucoamylase was found. the nucleotide sequence of this enzyme, designated tga, consists of 1,953 base pairs corresponding to a protein of 651 amino acid residues. the tga gene was subcloned and expressed in escherichia coli. enzymatic analyses showed that, like other glucoamylases, tga produced beta-d-glucose ...200111549021
physiology, biochemistry and taxonomy of deep-sea nitrile metabolising rhodococcus strains.a collection of nitrile-hydrolysing rhodococci was isolated from sediments sampled from a range of deep coastal, and abyssal and hadal trench sites in the nw pacific ocean, as part of our programme on the diversity of marine actinomycetes. nitrile-hydrolysing strains were obtained by batch enrichments on nitrile substrates with or without dispersion and differential centrifugation pre-treatment of sediments, and were recovered from all of the depths sampled (approximately 1100-6500 m). two isola ...200111759050
identification and properties of type i-signal peptidases of bacillus amyloliquefaciens.the use of bacillus amyloliquefaciens for enzyme production and its exceptional high protein export capacity initiated this study where the presence and function of multiple type i signal peptidase isoforms was investigated. in addition to type i signal peptidases sips(ba) [meijer, w.j.j., de jong, a., bea, g., wisman, a., tjalsma, h., venema, g., bron, s. & van dijl, j.m. (1995) mol. microbiol. 17, 621-631] and sipt(ba) [hoang, v. & hofemeister, j. (1995) biochim. biophys. acta 1269, 64-68] whi ...200211856304
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