Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.2.1.26 (invertase)
4,927 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

An enzymatic spectrophotometric assay for the determination of sucrose in unextracted samples of serum and urine was developed. The method entailed the coupling of invertase-catalyzed sucrose hydrolysis with a fructose dehydrogenase-catalyzed oxidation of the liberated fructose. The latter reaction generated reducing equivalents that were transferred to a tetrazolium salt with a concomitant increase in absorbance at 570 nm. The assay, which was carried out in microtiter plates, had a minimum detectable sucrose concentration of 0.03 mmol/liter and run-to-run and within-run coefficients of variation of 7.5 and 6.7%, respectively, and showed a good correlation with urine sucrose determination by GLC (r = 0.92). The assay range of 0.03-2.10 mmol/liter is suitable for the quantitation of serum sucrose following iv administration and for the quantitation of urine sucrose at basal levels and following the consumption of an oral test dose of sucrose. This method was used to analyze urine samples from a group of human subjects who consumed 20 g of sucrose for the assessment of gastroduodenal permeability. This convenient assay provides for the rapid and specific estimation of sucrose and has the potential to be used in a variety of manual, semiautomated, or automated formats.
...
PMID:Coupled enzymatic assay for the determination of sucrose. 902 15

Taurocholate transport was studied in brush border membrane vesicles (BBMV) isolated from chicken small (duodenum, jejunum, and ileum) and large (proximal cecum and rectum) intestines, using a rapid filtration technique. The purity of the BBMV was verified by the finding that the specific activity of sucrase (a brush border membrane enzyme marker) was severalfold greater in vesicles than corresponding values in mucosal homogenate. The functional integrity of isolated BBMV was evaluated by the uptake of D-glucose, which showed a transient increase in the presence of Na+. A Na+-dependence of taurocholate uptake was shown in BBMV prepared from ileum, cecum, and rectum, as taurocholate transport was transiently increased (accumulation) in the presence of a Na+ gradient between the external medium and intravesicular medium. The magnitude of the accumulation was similar among ileum, cecum, and rectum. In contrast, BBMV prepared from duodenum and jejunum did not show any Na+-dependent taurocholate transport, as the taurocholate uptake was not affected when a Na+ gradient was replaced by a K+ gradient. The use of taurochenodeoxycholate in the incubation medium inhibited Na+-dependent taurocholate transport in the ileum, cecum, and rectum. This study is the first to show the presence of a Na+-dependent bile salt transport in BBMV obtained from chicken ileum, proximal cecum, and rectum.
...
PMID:Taurocholate transport by brush border membrane vesicles from different regions of chicken intestine. 956 44

A potent natural alpha-glucosidase inhibitor called kotalanol has been isolated from an antidiabetic traditional Ayurvedic medicine, the roots and stems of Salacia reticulata Wight, through bioassay-guided separation. The structure of kotalanol was elucidated on the basis of chemical and physicochemical evidence to be the inner salt comprised of 1-deoxyheptosyl-3-sulfate anion and 1-deoxy-4-thio-D-arabinofuranosyl sulfonium cation. Kotalanol was found to show more potent inhibitory activity against sucrase than salacinol and acarbose.
...
PMID:Kotalanol, a potent alpha-glucosidase inhibitor with thiosugar sulfonium sulfate structure, from antidiabetic ayurvedic medicine Salacia reticulata. 973 18

We report the characterization of N-linked oligosaccharides on six foreign glycoproteins secreted from the methylotrophic yeast Pichia pastoris. These proteins included: a bacterial enzyme, Bacillus licheniformis alpha-amylase; three fungal enzymes, Saccharomyces cerevisiae invertase, Penicillium minioluteum dextranase, and Mucor pusillus aspartic protease; and two higher eukaryotic proteins, Boophilus microplus (tick) gut antigen and bovine enterokinase catalytic subunit. The carbohydrates on these proteins were observed to vary in size, with Man8GlcNAc2 and Man9GlcNAc2 structures being the most frequently observed species. Substantial amounts of shorter oligomannoside structures were present only on invertase, and longer structures (up to Man18GlcNAc2) were common on aspartic protease and enterokinase. Phosphorylated oligosaccharides were observed on one protein, aspartic protease. Unlike oligosaccharides on glycoproteins secreted from S. cerevisiae, no terminal alpha1,3-linked mannosylation was observed on any of the six P. pastoris-secreted proteins. Changing the growth and induction medium from a minimal salt-based medium to a molasses-based medium had little effect on the size of the oligomannosides. From these results, it is apparent that most foreign proteins secreted from P. pastoris are not subjected to the extensive mannosylation (hyperglycosylation) that commonly occurs in proteins secreted from S. cerevisiae.
...
PMID:Variation in N-linked oligosaccharide structures on heterologous proteins secreted by the methylotrophic yeast Pichia pastoris. 979 Aug 82

The three subunits of the nascent polypeptide-associated complex (alpha, beta1, beta3) in Saccharomyces cerevisiae are encoded by three genes (EGD2, EGD1, BTT1). We found the complex bound to ribosomes via the beta-subunits in a salt-sensitive manner, in close proximity to nascent polypeptides. Estimation of the molecular weight of the complex of wild-type cells and cells lacking one or two subunits revealed that the composition of the complex is variable and that as yet unknown proteins might be included. Regardless of the variability, a certain balance of the subunits has to be maintained: the deletion of one subunit causes downregulation of the remaining subunits at physiological growth temperature. Cells lacking both beta-subunits are unable to grow at 37 degrees C, most likely due to a toxic effect of the alpha-subunit. Based on in vitro experiments, it has been proposed that the function of mammalian nascent-polypeptide associated complexes (NAC) is to prevent inappropriate targeting of non-secretory nascent polypeptides. In vivo, however, the lack of NAC does not cause secretion of signal-less invertase in yeast. This result and the lack of a drastic phenotype of cells missing one, two or three subunits at optimal conditions (28 degrees C, YPD-medium) suggest either the existence of a substitute for NAC or that cells tolerate or 'repair' the damage caused by the absence of NAC.
...
PMID:Initial characterization of the nascent polypeptide-associated complex in yeast. 1021 98

The response to moderate salt stress of a Scytonema species isolated from a soil crust in the arid region of central Australia was studied. An increase in intracellular trehalose and sucrose concentrations was detected by NMR and HPLC analysis following salt stress, maximal amounts being produced by exposure to 150 mM NaCl after 48 h. When the organism was subsequently returned to normal growth conditions, the cellular concentrations of these solutes decreased. The biosynthesis of trehalose and sucrose was studied and found, in both cases, to involve both sugar phosphate synthase and phosphatase enzymes. The combined synthase activities and the individual phosphatase activities in cell extracts were increased by salt stress. Trehalose phosphorylase was the only catabolic enzyme detected for trehalose; neither trehalase nor phosphotrehalase activities could be detected. This is the first report of trehalose phosphorylase activity in cyanobacteria. Both trehalose and sucrose phosphorylase activities increased in salt-stressed cells, whereas the activity of invertase did not change.
...
PMID:Involvement of the compatible solutes trehalose and sucrose in the response to salt stress of a cyanobacterial Scytonema species isolated from desert soils. 1056 66

A major difficulty with isolating enzymatically or chemically released oligosaccharides from large-scale glycoprotein deglycosylation reactions is the time-consuming chromatography, desalting, and concentration steps required to prepare a glycan fraction of manageable proportions. To overcome these time and preparative chromatography equipment requirements, we have developed a rapid organic solvent precipitation/extraction procedure that allows sequential isolation of endo-beta-N-acetylglucosaminidase H (EC 3.2.1.96)-released high-mannose and hybrid, peptide-N(4)-(N-acetyl-beta-glucosaminyl) Asn amidase (EC 3.5.1. 52)-released complex, and beta-eliminated O-linked glycans without the need for intermediate chromatography, desalting, or concentration steps. The method involves precipitation of protein and released glycans at -20 degrees C in 80% acetone and extraction of the glycans from the pellet with 60% aqueous methanol after each deglycosylation step. Three pools of essentially salt- and detergent-free oligosaccharides (high-mannose/hybrid, complex, and O-linked) can be isolated in a high yield in 4 days with this protocol, which has been extensively tested using bovine RNase B, human bile salt-stimulated lipase expressed in Pichia pastoris, hen ovalbumin, bovine fetuin, bovine thyroglobulin, and several invertase preparations from wild-type and mutant yeast strains.
...
PMID:Selective organic precipitation/extraction of released N-glycans following large-scale enzymatic deglycosylation of glycoproteins. 1066 Apr 52

The non-pathogenic, dimorphic, ascomycetous yeast Arxula adeninivorans LS3 is halotolerant. It can grow in a minimal medium containing up to 20% NaCl. The growth parameters are only weakly influenced by 10% NaCl. However, NaCl in a concentration higher than 10% causes a decrease in the specific growth rate, a longer adaptation phase and a lower cell count in the stationary growth phase. Concentrations of glycerol and trehalose, which differed 100-fold in magnitude in a salt free medium, are also influenced differently by salt. NaCl induces accumulation of intracellular glycerol in exponentially growing cells but a reduced concentration of intracellular trehalose in stationary cells. Transcripts of the genes ARFC3, encoding a component of the replication factor C, and GAA, encoding a secretory glucoamylase, can be detected only in cells cultured in media with NaCl concentrations below 10%. Furthermore, NaCl in high concentration reduces the level of secreted proteins including glucoamylase end invertase.
...
PMID:Halotolerance of the yeast Arxula adeninivorans LS3. 1095 59

We investigated the salt tolerance of transgenic tobacco, in which yeast invertase is expressed in the apoplastic (Apo-Inv) spaces. Whereas photosynthetic activities in wild-type tobacco in light were inhibited under salt stress, transgenic Apo-Inv tobacco maintained constant photosynthetic activities. The physical appearance of plants under salt stress also indicates that yeast invertase expression in the apoplastic space is beneficial for inducing salt tolerance. Apo-Inv tobacco had a much higher osmotic pressure increase in the cell sap than did wild-type tobacco under this type of stress. The physiological importance of sucrose metabolism under salt stress is discussed.
...
PMID:Improved salt tolerance of transgenic tobacco expressing apoplastic yeast-derived invertase. 1123 May 81

Three different isoforms of invertases have been detected in the developing internodes of barley (Hordeum vulgare). Based on substrate specificities, the isoforms have been identified to be invertases (beta-fructosidases EC 3.2.1.26). The soluble (cytosolic) invertase isoform can be purified to apparent homogeneity by diethylaminoethyl cellulose, Concanavalin-A Sepharose, organo-mercurial Sepharose, and Sephacryl S-300 chromatography. A bound (cell wall) invertase isoform can be released by 1 molar salt and purified further by the same procedures as above except omitting the organo-mercurial Sepharose affinity chromatography step. A third isoform of invertase, which is apparently tightly associated with the cell wall, cannot be isolated yet. The soluble and bound invertase isoforms were purified by factors of 60- and 7-fold, respectively. The native enzymes have an apparent molecular weight of 120 kilodaltons as estimated by gel filtration. They have been identified to be dimers under denaturing and nondenaturing conditions. The soluble enzyme has a pH optimum of 5.5, Km of 12 millimolar, and a Vmax of 80 micromole per minute per milligram of protein compared with cell wall isozyme which has a pH optimum of 4.5, Km of millimolar, and a Vmax of 9 micromole per minute per milligram of protein.
...
PMID:Purification and characterization of soluble (cytosolic) and bound (cell wall) isoforms of invertases in barley (Hordeum vulgare) elongating stem tissue. 1153 66


<< Previous 1 2 3 4 5 6 7 8 9 Next >>

---