Search results for "Enzyme Assay"
showing 10 items of 155 documents
Purification and characterization of leucine aminopeptidase from kidney bean cotyledons
1992
A leucine aminopeptidase (EC 3,4,11.1) was purified from cotyledons of resting kidney beans (Phaseolus vulgaris L. cv. Processor) by acidic extraction, ammonium sulfate fractionation and chromatography on DEAE-Sephacel, Sephacryl S-300, Mono Q HPLC and Superose HPLC columns. The yield of the 317-fold purified enzyme was 9%. On gel filtrations on Sephacryl S-300 and Superose HPLC the elution volumes of the enzyme corresponded to an M, of 360 000. The enzyme gave one band on native gel electrophoresis and an electrophoretic titration in an immobilized pH gradient gave a single curve with a pI of 4.8. Two bands were observed in an SDS-gel electrophoresis with Mr values of 58 000 and 60 000 bot…
Effects of protein on retention of ADH enzyme activity encapsulated in trehalose matrices by spray drying
2008
The retention of the enzymatic activity of alcohol dehydrogenase (ADH) on spray drying was examined under various drying conditions. Trehalose was used in the formulation of feed stocks for the spray drying. The retention of ADH activity was dependent on the concentration of ADH in the feed solution. The addition of other proteins, such as bovine serum albumin and β-lactoglobulin, exhibited an additional improvement of the retention of ADH activity. The inlet and outlet temperature of the drying air was another key factor for the enzyme activity on the spray drying. The surface morphology of the spray-dried particle was changed drastically with the addition of proteins.
Deoxyribonucleases in Herpes simplex Virus Type 1 and 2 Infected Primary Rabbit Kidney Cells
1980
Abstract In primary rabbit kidney cells infected with herpes simplex virus four different neutral deoxyribonuclease activities can be detected by means of the deoxyribonuclease assay in DNA-containing polyacrylamide gels following their separation by discelectrophoresis. The method is suitable to follow independently the change in each activity of the different enzymes using only about 5 × 105 cells for each assay during the time-course of infection. Under these conditions one enzyme activity is constant, two disappear while the activity of a fourth one present only in infected cells, increases.
The Effect of Phosphinothricin (Glufosinate) on Glutathione Synthesis in Plants
1990
The inhibitory effect of DL-phosphinothricin (glufosinate) on glutathione synthesis was studied in vivo and in vitro. The influence of phosphinothricin on γ-glutamylcysteine synthetase was compared with the already known effects of l-buthionine sulfoximine and l-methionine sulfoximine. The results showed that phosphinothricin and buthionine sulfoximine are inhibitors of γ-glutamylcysteine synthetase of plants. With both substances the enzyme was inhibited by 50 % at a concentration of 7 . 10−4M (pI50 = 3.15). Methionine sulfoximine reduced the enzyme activity by 50% at 5 . 10−2 M (pI50 = 1.30). It is discussed that the target enzyme of phosphinothricin is the glutamine synthetase whereas th…
Glutathione-dependent defence system and monooxygenase enzyme activities in Arctic charr Salvelinus alpinus (L.) exposed to ozone
2000
Abstract One of the major obstacles of the increasing usage of ozone in aquaculture is the lack of relevant risk assessment in culture conditions. Before the apparent advantages of ozonation can be utilised efficiently, the safety margins and biological basis of ozone toxicity should be assessed. In this research, 1-year-old Arctic charr ( Salvelinus alpinus (L.)) were exposed to an ozone concentration high enough to inactivate Aeromonas sp. bacteria in freshwater, but too low to be directly lethal to the fish themselves. The effects of ozone exposure on the activity of glutathione-dependent antioxidant enzymes and monooxygenase reactions were studied in blood and in liver. The fish were ac…
Inhibition of herpesvirus DNA synthesis by 9-beta-D-arabinofuranosyladenine in cellular and cell-free systems.
1977
9-beta-D-Arabinofuranosyladenine 5'-triphosphate (ara-ATP) is an inhibitor both of DNA polymerase-alpha and -beta from noninfected rabbit kidney cells and of the DNA-dependent DNA polymerase induced by herpes simplex virus Type 1 (strain IES). The studies were performed with partially purified enzymes, and each of the different polymerase preparations contained only one DNA-dependent DNA polymerase species. These enzymes were inhibited in a competitive manner. The HSV-induced DNA-dependent DNA polymerase was 39-fold more sensitive to ara-ATP than was cellular DNA polymerase-beta and 116-fold more sensitive than cellular DNA polymerase-alpha. The affinity of the HSV-induced enzyme for ara-AT…
Immediate early response of the marine sponge Suberites domuncula to heat stress: Reduction of trehalose and glutathione concentrations and glutathio…
1997
The marine sponge Suberites domuncula was used to identify early markers for thermal stress. Cubes from sponges have been kept for 30 min at 31°C (10 °C higher than the ambient temperature). After this treatment the sponge cubes were kept again at 21°C. To demonstrate that the animals reacted to the elevated temperature, the expression of heat shock protein (HSP) was determined. Using an antibody raised against HSP70, it was found by Western blotting that the animals specifically express a 45 kDa polypeptide after heat treatment. It was shown that even after 10 min of heat treatment the steady-state concentration of trehalose drops by 40% from a base level of 13 nmol/mg protein. The activit…
Biochemical Characterization of an Acetylcholine-hydrolyzing Enzyme from Bean Seedlings.
1980
An acetylcholine hydrolyzing enzyme was prepared and purified (40 times) from dwarf bean hypocotyl hooks. The purity of the enzyme was proved by polyacrylamide gel electrophoresis. The molecular weight of the enzyme was determined to be 65,000 daltons. Enzyme activity was the highest at pH 8.0 and between 30 and 36 C. The enzyme had an apparent affinity constant (K(m)) for acetylcholine of 460/micromolar. The affinity for substrate analogs increased from butyrylthiocholine to propionylthiocholine to acetylthiocholine. The enzyme activity was inhibited by choline, neostigmine, physostigmine, manganese, and calcium. Magnesium had no influence on the enzyme activity. We conclude that the enzym…
The Effect of Light on the Growth of Pea Plants and the Subsequent Influence in Shikimate Oxidoreductase (EC 1.1.1.25) Activity
1981
Summary Pea plants were cultured in white light, red and far red light, and in the dark during a period of three weeks. At several states of development we investigated the activity of the enzyme shikimate oxidoreductase, the amount of fresh and dry matter, and the contents of protein in stem, leaves, cotyledons, and roots. The enzyme activity was found to be distributed organ-specifically and uninfluenced by the phytochrome system, but it was significantly depressed in plants grown in the dark compared to plants grown in white light. Enzyme activity occurred also in non photosynthetic plants. Regarding the different light conditions the activity of shikimate oxidoreductase was found to cor…
Endogenous Role of Microsomal Epoxide Hydrolase
2005
The specific activities of microsomal epoxide hydrolase with 16α,17α-epoxyandrosten-3-one (androstene oxide) as substrate were measured in various metabolically important and in various steroidogenic organs of the male and female rat and compared with the activities of 16α,17α-epoxyestratrienol (estroxide) and benzo[a]pyrene 4,5-oxide. Androstene oxide was an exceptionally good substrate. The specific activities differed widely between organs but the ratio of the activities towards these substrates was constant in all organs investigated. The ratios compared to benzo[a]pyrene 4,5-oxide were 2.5 for estroxide, and 8.6 for androstene oxide. The ontogenetic development of specific epoxide hydr…