Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.4.1.2 (glutamate dehydrogenase)
 4,380 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A method was developed for extracting enzymes from micro-organisms closely associated with ammonia-treated straw (NH3-S) that had been incubated in nylon bags in the rumen. Incubation of washed straw with 125 ml carbon tetrachloride/l and 20 micrograms lysozyme/ml for 3 h at 37 degrees gave carboxymethylcellulase (EC 3.2.1.4; CMCase) and NAD-linked glutamate dehydrogenase (EC 1.4.1.2; GDH) activities greater than those extracted by sonication. GDH associated with NH3-S increased with incubation time and was highest in sheep receiving a high-barley diet. Particle-bound CMCase activity reached a peak between 16 and 24 h and declined thereafter. Particle-bound GDH activity showed no correlation with dry matter (DM) degradation in the rumens of sheep fed on a range of diets. In contrast, CMCase activity after 24 h was highly correlated with DM degradability of the same samples at 24 h (r 0.98) and 48 h (r 0.94). It was concluded that GDH and CMCase can be used as indices of the total population of colonizing rumen micro-organisms and of the fibre-degrading population respectively, and that these enzymes can therefore be used to assess rapidly and with great sensitivity variations in the rumen environment that affect the rate of fibre breakdown.
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PMID:Use of particle-bound microbial enzyme activity to predict the rate and extent of fibre degradation in the rumen. 303 99

Mesophyll cells (MCs) and bundle-sheath cells (BSCs) of leaves of the C4 plant maize (Zea mays L.) were separated by cellulase digestion to determine the relative proportion of the glutamine synthetase (GS; EC 6.3.1.2) or the NADH-glutamate dehydrogenase (GDH; EC 1.4.1.2) isoforms in each cell type. The degree of cross-contamination between our MC and BSC preparations was checked by the analysis of marker proteins in each fraction. Nitrate reductase (EC 1.6.6.1) proteins (110 kDa) were found only in the MC fraction. In contrast, ferredoxin-dependent glutamate synthase (Fd-GOGAT; EC 1.4.7.1) proteins (160 kDa) were almost exclusively present in the BSC fraction. These results are consistent with the known intercellular distribution of nitrate reductase and Fd-GOGAT proteins in maize leaves and show that the cross-contamination between our MC and BSC fractions was very low. Proteins corresponding to cytosolic GS (GS-1) or plastidic GS (GS-2) were found in both the MC and BSC fractions. While equal levels of GS-1 (40 kDa) and GS-2 (44 kDa) polypeptides were present in the BSC fraction, the GS-1 protein level in the MC fraction was 1.8-fold higher than the GS-2 protein pool. Following separation of the GS isoforms by anion-exchange chromatography of MC or BSC soluble protein extracts, the relative GS-1 activity in the MC fraction was found to be higher than the relative GS-2 activity. In the BSC fraction, the relative GS-1 activity was very similar to the relative GS-2 activity. Two isoforms of GDH with apparent molecular weights of 41 kDa and 42 kDa, respectively, were detected in the BSC fraction of maize leaves. Both GDH isoenzymes appear to be absent from the MC fraction. In the BSCs, the level of the 42-kDa GDH isoform was 1.7-fold higher than the level of the 41-kDa GDH isoform. A possible role for GS-1 and GDH co-acting in the synthesis of glutamine for the transport of nitrogen is discussed.
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PMID:Glutamine synthetase and glutamate dehydrogenase isoforms in maize leaves: localization, relative proportion and their role in ammonium assimilation or nitrogen transport. 1114 64

Etiolated pea (Pisum sativum) epicotyls synthesize a buffer-soluble cellulase (cellulase A) and a salt-soluble cellulase (cellulase B) (EC 3.2.1.4) after treatment with high (0.5%) auxin levels. Only cellulase A increased in activity after treatment with low (0.005%) auxin. Cellulase A was released into the supernatant after homogenization of tissue in dilute buffer (buffer-soluble), had a pH optimum at 5.5, was relatively thermostable, and its activity was inhibited by NaCl. Cellulase B was released by 1 m NaCl (salt-soluble) from excised tissue segments or from the insoluble residue remaining after removal of the buffer-soluble form. It had a pH optimum at 7.0, was thermolabile, and required salt for maximum activity. When subjected to polyacrylamide gel electrophoresis, the cellulase fraction released by NaCl from excised segments showed two bands of cellulase activity compared to several for the buffer-soluble fraction. Electrophoretic analysis of the buffer and salt-soluble fractions for marker enzymes indicated the presence of malate dehydrogenase activity in all fractions and glutamate dehydrogenase activity in the buffer-soluble fraction only.Exposure of intact pea epicotyls to 70 mul/l of ethylene gas for 3 days did not affect cellulase A activity, but caused a 5-fold increase in cellulase B activity (enzyme released by salt from the buffer-insoluble residue). We concluded that ethylene and auxin generate different forms of cellulase.
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PMID:Extraction and partial characterization of cellulases from expanding pea epicotyls. 1665 14

In a switch-over experiment, eight male animals, four each of sheep and goats of local breeds with mean body weight of 26. 8 +/- 2.0 and 30.0 +/- 2.1 kg, were fed Dichanthium annulatum (DA) grass and four browse species viz. Helictris isora, Securengia virosa, Leucaena leucocephala (LL) and Hardwickia binnata (HB) in four feeding trials to assess their supplementary effect on activity of rumen enzymes. The sheep and goats were offered DA grass with individual browse in 75:25 and 50:50 proportions, respectively, for more than 3 months during each feeding trial, and rumen liquor samples were collected twice at 0 and 4 h post feeding after 60 and 90 days of feeding. Glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and glutamate dehydrogenase (GDH) enzymes were determined in the bacteria and protozoa fractions of rumen liquor, while cellulase enzyme activity was measured in mixed rumen liquor. LL and HB had the highest and lowest contents of CP, while fibre contents were lower in early than later browse leaves. Supplementation of browse leaves significantly (P < 0.05) affect the specific activity of GDH enzyme in bacteria fraction of rumen liquor of animal species, while GDH activity was similar in protozoa fraction of rumen liquor of sheep and goats on all DA grass-browse-supplemented diets except DA-HB (42.8 units/mg protein), where activity was significantly (P < 0.05) low. Specific activities of GOT and GPT enzymes in both bacteria and protozoa fractions of rumen liquor differ significantly (P < 0.05) due to supplementation of browse leaves to DA grass. Browse leaves significantly (P < 0.05) affect the cellulase enzyme activity in animal rumen liquor, being highest on DA-LL (193.4) and lowest on DA-HB diet (144.8 microg sugar/mg protein). Goat exhibited higher activities of GOT and GPT than sheep in both bacteria and protozoa fraction of rumen liquor, while cellulase activity was similar between the animal species on the grass-browse leaves diets. Results indicate that browse leaves supplementation affect the enzyme activities of sheep and goats rumen, while the goats rumen liquor had higher activities of GOT, GPT and GDH enzyme than sheep.
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PMID:Effect of tropical browse leaves supplementation on rumen enzymes of sheep and goats fed Dichanthium annulatum grass-based diets. 2035 30

Escherichia coli is a common host for recombinant protein production in which production titers are highly dependent on the employed expression system. Promoters are thereby a key element to control gene expression levels. In this study, a novel PLICable promoter toolbox was developed which enables in a single cloning step and after a screening experiment to identify out of ten IPTG-inducible promoters (T7, A3, lpp, tac, pac, Sp6, lac, npr, trc and syn) the most suitable one for high level protein production. The target gene is cloned under the control of different promoters in a single and efficient cloning step using the ligase-free cloning method PLICing (phosphorothioate-based ligase-independent gene cloning). The promoter toolbox was firstly validated using three well producible proteins (a cellulase from a metagenome library, a phytase from Yersinia mollaretii and an alcohol dehydrogenase from Pseudomonas putida) and then applied to two enzymes (3D1 DNA polymerase and glutamate dehydrogenase mutant) which are poorly produced in E. coli. By applying our PLICable pET-promoter toolbox, the authors were able to increase production by two-fold for 3D1 DNA polymerase (lac promoter) and 29-fold for glutamate dehydrogenase mutant H52Y (trc promoter).
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PMID:Screening through the PLICable promoter toolbox enhances protein production in Escherichia coli. 2775 30