SEARCH · Search PubMed

Results for “Trehalase”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

SourceResearch connectionFrom yearThrough year

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 163 records · Page 9Linked to original sources

PubMed1980

Effect of high pressure on the heat activation in vivo of trehalase in the spores of Phycomyces blakesleeanus.

The effect of pressure on the heat activation in vivo of trehalase in the spores of Phycomyces blakesleeanus has been investigated in order to obtain information about the molecular mechanism of the activation. For a protein conformational change directly induced in the enzyme by the heat treatment an upward shift with about 2-6 K/1000 atm (1.013 X 10(5) kPa) is to be expected in the moderate high-pressure region. On the other hand, for a phospholipid phase transition causing the activation, a continuous upward shift with about 20 K/1000 atm is to be expected. For trehalase activation we find a continuous upward shift of the activation temperature with about 5-9 K/1000 atm. The denaturation of trehalase, which occurs at slightly higher temperatures, is influenced by pressure completely as expected for a protein conformational change. The application of high pressure during spore heat activation makes it possibe to break the dormancy of the spores without concomitant activation of trehalase.

Alcohols↗

PubMed1999

Stress tolerance in doughs of Saccharomyces cerevisiae trehalase mutants derived from commercial Baker's yeast.

Accumulation of trehalose is widely believed to be a critical determinant in improving the stress tolerance of the yeast Saccharomyces cerevisiae, which is commonly used in commercial bread dough. To retain the accumulation of trehalose in yeast cells, we constructed, for the first time, diploid homozygous neutral trehalase mutants (Deltanth1), acid trehalase mutants (Deltaath1), and double mutants (Deltanth1 ath1) by using commercial baker's yeast strains as the parent strains and the gene disruption method. During fermentation in a liquid fermentation medium, degradation of intracellular trehalose was inhibited with all of the trehalase mutants. The gassing power of frozen doughs made with these mutants was greater than the gassing power of doughs made with the parent strains. The Deltanth1 and Deltaath1 strains also exhibited higher levels of tolerance of dry conditions than the parent strains exhibited; however, the Deltanth1 ath1 strain exhibited lower tolerance of dry conditions than the parent strain exhibited. The improved freeze tolerance exhibited by all of the trehalase mutants may make these strains useful in frozen dough.

Carbon Dioxide↗

PubMed2002

Identification of Candida glabrata by a 30-second trehalase test.

Rapid (30-s) trehalase tests done with material from colonies of 482 yeasts suspended in a drop of trehalose solution on a commercially supplied glucose test strip were positive for 225 (99.1%) of 227 Candida glabrata isolates grown on either of two differential media, Candida ID medium or CandiSelect medium. The test was positive for only 3 (1.2%) and 12 (4.7%) of 255 isolates of other medically important yeast species grown on the same two media, respectively. A rapid maltase test done with a subset of 255 yeast isolates was negative for all but 1 of 64 trehalase-positive C. glabrata isolates, raising the specificity of the rapid testing for C. glabrata to 98.4 to 100%, depending on the isolation medium used. Rapid trehalase and maltase tests done independently in two laboratories with 217 yeast isolates showed sensitivities of 96.0 to 98.0% and specificities of 98.2 to 99.4% for identification of C. glabrata from colonies grown on Candida ID medium. The specificity was much lower because of frequent false-positive trehalose test results when the source of colonies was Sabouraud agar formulated with 4% glucose. We conclude that direct recognition of C. albicans as blue colonies on Candida ID isolation medium coupled with the performance of the 30-s trehalase and maltase tests for C. glabrata among the white colonies on this medium will allow the rapid presumptive identification of the two yeast species most commonly encountered in clinical samples.

Candida glabrata↗

PubMed1988

Trehalase deficiency in Greenland.

Small-intestinal surgical biopsy specimens from 97 adult Greenlanders showed an incidence of trehalase deficiency in at least 8%. Trehalose tolerance tests performed in three individuals with low trehalase activity (6 IU/g protein) showed no increase in the blood glucose concentration. Three trehalase-deficient patients had lactase deficiency too. Trehalase deficiency is not considered to have any significance from a nutritional point of view.

Adult↗

PubMed1997

Gene expression of trehalase during post-dormant development of the brine shrimp, Artemia: comparison of the two species.

Based on a homology screening approach, two degenerate oligonucleotides were employed as primers in a polymerase chain reaction to amplify a fragment of DNA encoding trehalase with a template of cDNA derived from embryos of American Artemia. Sequence analysis revealed that the fragment was composed of 228 bp comprising 76 amino acids, and highly homologous to trehalases of Tenebrio molitor (mealworm beetle), rabbit, Caenorhabditis elegans, Bombyx mori (silkworm) and Escherichia coli treA and treF (58-38%, in order of description). This fragment was used as a hybridization probe. A Northern blot analysis on American Artemia showed three transcripts of 5.0, 2.7 and 2.2 kb, and the two larger transcripts were also detected in Chinese Artemia. The developmental profile of the gene expression and the trehalase activity suggest that the transcripts of 5.0 and (or) 2.7 kb in both Artemia may be directly or indirectly related to translation of the trehalase. A Southern blot analysis on both Artemia suggested the existence of two highly homologous genes or one gene having an intron within the region where the probe binds in their haploid genome.

Amino Acid Sequence↗

PubMed1991

Activation of yeast trehalase by heat shock.

1. Activation of Saccharomyces cerevisiae trehalase by heat shock was shown in all strains tested, including mutants in which the response to a glucose signal was absent. A low concentration of cAMP favored the response as seen in 2nd log cells or in ras2 and cyr1ts mutant strains. The heat shock effect upon trehalase activity was not observed under conditions of catabolite repression. 2. Neither hexokinase PII nor the heat shock protein hsp26 seemed to be involved in the activation of trehalase by heat shock. However, mutant strains deleted in the polyubiquitin gene showed only a 2-fold activation of the enzyme while in control strains a 5- to 7-fold irreversible activation was observed. 3. An alternative mechanism of trehalase activation by removal of an inhibitor through ligation with ubiquitin is discussed. Activation by cAMP-independent phosphorylation is also considered.

Culture Media↗

PubMed1991

Modulation of trehalase activity in two insect cell lines by virus infection and trehalose.

Trehalase (EC 3.2.1.28), an important glycosidase involved in regulating trehalose levels and metabolic energy in insects, was measured in cell lines from fall army worm, Spodoptera frugiperda and salt marsh caterpillar, Estigmene acrea, treated with either glucose or trehalose in the presence or absence of Tipula Iridescent Virus (TIV), a cytoplasmic deoxyribovirus. In medium containing 15-35 mM trehalose, both of these cells increased their trehalase activities by 4.5 to 8x the basal levels from cells in glucose medium. Trehalase activity was rapidly reduced after cells were exposed to TIV. Maximum loss in activity (70-90%), occurring about the same time as peak viral DNA synthesis, was significantly delayed when cells were pre-incubated with 30 mM trehalose. These experiments demonstrate the potential utility of trehalase as a marker for monitoring stresses induced by viral infection and changes in nutrition.

Animals↗

PubMed1981

Trehalose Toxicity in Cuscuta reflexa: CORRELATION WITH LOW TREHALASE ACTIVITY.

A toxic effect of alpha,alpha-trehalose in an angiospermic plant, Cuscuta reflexa (dodder), is described. This disaccharide and its analogs, 2-aminotrehalose and 4-aminotrehalose, induced a rapid blackening of the terminal region of the vine which is involved in elongation growth. From the results of in vitro growth of several angiospermic plants and determination of trehalase activity in them, it is concluded that the toxic effect of trehalose in Cuscuta is because of the very low trehalase activity in the vine. As a result, trehalose accumulates in the vine and interferes with some process closely associated with growth. The growth potential of Lemna (a duckweed) in a medium containing trehalose as the carbon source was irreversibly lost upon addition of trehalosamine, an inhibitor of trehalase activity. It is concluded that, if allowed to accumulate within the tissue, trehalose may be potentially toxic or inhibitory to higher plants in general. The presence of trehalase activity in plants, where its substrate has not been found to occur, is envisaged to relieve the plant from the toxic effects of trehalose which it may encounter in soil or during association with fungi or insects.

Journal Article↗

PubMed1967

Mechanisms of protection of trehalase against heat inactivation in Neurospora.

The half-life of trehalase and invertase at 65 and 60 C was found to be much greater when intact ascospores of Neurospora tetrasperma were heated, as compared with extracts. By contrast, no protection was afforded these enzymes when they were heated in intact conidia and mycelium of N. crassa or N. tetrasperma. The protective effect of ascospores for trehalase was further investigated by heating ascospore extracts before and after dialysis. The removal of small molecules by dialysis lowered the heat resistance of trehalase significantly in such extracts. When the dialysate from extracts of mycelium, conidia, or ascospores was added to dialyzed enzyme extracts, that from ascospores was by far the most active. However, the same dialysates had only a small protective effect on invertase. The addition of ashed dialysates did not protect trehalase, and trehalose and glucose protected less effectively than the dialysate.

Dialysis↗

PubMed2001

Determination of 1,5-anhydro-D-glucitol on the basis of its inhibitory effect on trehalase activity.

1,5-Anhydro-D-glucitol (1,5-AG) was found to inhibit trehalase and trehalose phosphorylase activities competitively, because of its structural similarity with D-glucose. Trehalase from Nocardia sp., one of the most 1,5-AG-sensitive enzymes, was used in the determination of 1,5-AG concentration, which is a useful marker for the diagnosis of diabetes. A good linear relationship was observed between 1,5-AG concentration in the range of 0.02 to 1.0 mM and the extent of trehalase inhibition by 1,5-AG. The 1,5-AG concentration range could be determined by estimating enzymatically the amount of the reaction product, D-glucose, produced by the trehalase.

Journal Article↗

PubMed1997

Determination of trehalose by flow injection analysis using immobilized trehalase.

A new method for the determination of trehalose by flow injection analysis (FIA) is described. The basic principle is the hydrolysis of the disaccharide trehalose into its monomer d-glucose by trehalase, a periplasmic enzyme of Escherichia coli. d-glucose is quantified spectrophotometrically after reaction with hexokinase and glucose-6-phosphate dehydrogenase. Trehalase is prepared by osmotic shock from a recombinant E. coli strain and precipitated with ammonium sulfate. The enzyme is immobilized on VA-Epoxy Biosynth from Riedel-de-Haën. The immobilization rate is about 60%. The FIA signals show a nonlinear dependence on the trehalose concentration. The resulting curve corresponds to a second-order polynomial that serves as a calibration function for test samples. Immobilized trehalase was used during a period of 4 months without any loss of suitability. Several samples of fermentation broth were tested. The results are verified by HPLC. Within an interval of 2 to 10 g/L trehalose the recovery is about 100-120% with a precision of 7% (coefficient of variation).

Autoanalysis↗

PubMed1982

Localization of trehalase in vacuoles and of trehalose in the cytosol of yeast (Saccharomyces cerevisiae).

Protoplasts of Saccharomyces cerevisiae synthesized and degraded trehalose when they were incubated in a medium containing traces of glucose and acetate. Such protoplasts were gently lyzed by the polybase method and a particulate and soluble fraction was prepared. Trehalose was found in the soluble fraction and the trehalase activity mostly in the particulate fraction which also contained the vacuoles besides other cell organelles. Upon purification of the vacuoles, by density gradient centrifugation, the specific activity of trehalase increased parallel to the specific content of vacuolar markers. This indicates that trehalose is located in the cytosol and trehalase in the vacuole. It is suggested that trehalose, in addition to its role as a reserve may also function as a protective agent to maintain the cytosolic structure under conditions of stress.

Cell Compartmentation↗

PubMed1988

Differential location of regulatory and nonregulatory trehalases in Candida utilis cells.

The isolation of vacuoles by density gradient centrifugation of protoplast lysates from Candida utilis cells showed a high specific activity for nonregulatory trehalase in vacuoles whereas the regulatory trehalase activatable by phosphorylation behaves as a cytoplasmic enzyme. The vacuolar trehalase is a glycoprotein that can be precipitated by Con A-Sepharose. Treatment of this enzyme with endo H reduced its reactivity with the lectin without loss of enzyme activity and decreased its apparent molecular weight by gel filtration.

Candida↗

PubMed1979

Effects of cortisone and thyroxine on intestinal trehalase activity in infant mouse.

Cortisone acetate (25 microgram/g b.wt/day) administration to 8-day-old suckling mice induces a premature increase of trehalase activity along the entire small intestine. On the other hand, thyroxine (1 microgram/g b.wt/day) in unable to provoke a precocious increase of trehalase activity. Trehalase appears to be the only brush border membrane disaccharidase controlled solely by glucocorticoid hormones during the postnatal maturation of the intestine.

Animals↗

PubMed1993

The interaction of Saccharomyces cerevisiae trehalase with membranes.

Plasma membranes isolated from cells of Saccharomyces cerevisiae previously submitted to a heat-shock showed a 10-fold increase in membrane-bound trehalase activity. Trehalase was purified to a high specific activity and was shown to be inhibited by glucose 6-phosphate and by the addition of a neutral phospholipid-like surfactant. Purified trehalase binds spontaneously to egg phosphatidylcholine small unilamellar vesicles, when in its active, phosphorylated form. When the enzyme was treated with alkaline phosphatase no binding was observed. The significance of this reversible binding for the control of trehalose metabolism in yeast cells is still unknown.

Cell Membrane↗

PubMed1978

Purification and properties of trehalase from the thermophilic fungus Humicola lanuginosa.

Trehalase (alpha,alpha-Trehalose glucohydrolase, EC 3.2.1.28) was partially solubilized from the thermophilic fungus Humicola lanuginosa RM-B, and purified 184-fold. The purified enzyme was optimally active at 50 degrees C in acetate buffer at pH 5.5. It was highly specific for alpha,alpha-trehalose and had an apparent Km = 0.4 mM at 50 degrees C. None of the other disaccharides tested either inhibited or activated the enzyme. The molecular weight of the enzyme was around 170 000. Trehalase from mycelium grown at 40 and 50 degrees C had similar properties. The purified enzyme, in contrast to that in the crude-cell free extract, was less stable. At low concentration, purified trehalase was afforded protection against heat-inactivation by "protection against heat-inactivation by "protective factor(s)" present in mycelial extracts. The "protective factor(s)" was sensitive to proteolytic digestion. It was not diffusible and was stable to boiling for at least 30 min. Bovine serum albumin and casein also protected the enzyme from heat-inactivation.

Dialysis↗

PubMed1996

Calystegine B4, a novel trehalase inhibitor from Scopolia japonica.

GLC-MS analysis has been developed for screening plants of the family Solanaceae for new calystegines. GLC-MS analyses of the extract of Scopolia japonica showed the presence of a new tetrahydroxy-nor-tropane alkaloid in addition to the known calystegines A3, A5, B1, B2, B3, and C1. We gave this new alkaloid the trivial name calystegine B4. The structure of calystegine B4 was determined as 1 alpha, 2 beta, 3 alpha, 4 alpha-tetrahydroxy-nor-tropane from a variety of NMR spectral data. Calystegines B1, B2, and C1 are potent competitive inhibitors with Ki values ranging from 10(-6) to 10(-7) M for almond beta-glucosidase, while calystegine B4 inhibited this enzyme in a competitive manner, with a Ki value of 7.3 microM. Calystegine B2 is also a potent inhibitor of green coffee bean alpha-galactosidase, whereas calystegine B4 exhibited no significant activity for this enzyme. Among rat intestinal glycosidases, only trehalase was potently inhibited by calystegine B4, with an IC50 value of 9.8 microM. Furthermore, calystegine B4 potently inhibited pig kidney trehalase in a competitive manner, with a Ki value of 1.2 microM, but it was almost inactive against yeast and fungal trehalases.

Alkaloids↗

PubMed1995

Phenotypic features of trehalase mutants in Saccharomyces cerevisiae.

In the yeast Saccharomyces cerevisiae, some studies have shown that trehalose and its hydrolysis may play an important physiological role during the life cycle of the cell. Recently, other studies demonstrated a close correlation between trehalose levels and tolerance to heat stress, suggesting that trehalose may be a protectant which contributes to thermotolerance. We had reported lack of correlation between trehalose accumulation and increase in thermotolerance under certain conditions, suggesting that trehalose may not mediate thermotolerance [Nwaka, S., et al. (1994) FEBS Lett. 344, 225-228]. Using mutants of the trehalase genes, NTH1 and YBR0106, we have demonstrated the necessity of these genes in recovery of yeast cells after heat shock, suggesting a role of these genes in thermotolerance (Nwaka, S., Kopp, M., and Holzer, H., submitted for publication). In the present paper, we have analysed the expression of the trehalase genes under heat stress conditions and present genetic evidence for the 'poor-heat-shock-recovery' phenotype associated with NTH1 and YBR0106 mutants. Furthermore, we show a growth defect of neutral and acid trehalase-deficient mutants during transition from glucose to glycerol, which is probably related to the 'poor-heat-shock-recovery' phenomenon.

Gene Expression Regulation, Fungal↗