Purification and some properties of isocitrate dehydrogenase of a blowfly Aldrichina grahami

• 1.1. NADH-dependent isocitrate dehydrogenase has been purified 110-fold from the crude extract of the flight muscle mitochondria of Aldrichina grahami.
• 2.2. The purification procedure involved Triton X-100 treatment of isolated mitochondria, column chromatography on DEAE-cellulose, Affi-gel blue, and P-cellulose.
• 3.3. The purified enzyme was homogeneous by criteria of the polyacrylamide gel electrophoresis.
• 4.4. The enzyme of the blowfly contains more acidic amino acids and less hydrophobic amino acids than that of pig heart.
• 5.5. The molecular weight was determined to be 330,000 daltons. The subunit construction differs from ghat of mammalian isocitrate dehydrogenase.

     

Anaerobic metabolism in sublittoral living Mytilus galloprovincialis in the mediterranean—IV. Role of amino acids in adaptation to low salinities during anaerobiosis and aerobiosis

• 1.1. When Mytilus galloprovincialis were transferred from 38 to 19%. sea water (S), the metabolism became anaerobic for at least 8 hr. After 24 hr the animals were entirely aerobic again.
• 2.2. Upon transfer to 19%. S, the total free amino acid concentration in haemolymph doubled within 4 hr, remaining nearly constant thereafter, up to 48 hr.
• 3.3. In the posterior adductor muscle a strong decrease of alanine and glycine occurred at 48 hr exposure to 19%. S, and a smaller decrease of glutamate; taurine remained relatively constant. When transferred again to 38%. S after 14 days, a strong overcompensation occurred in the concentrations of alanine and proline, and a smaller overcompensation in those of threonine and serine.
• 4.4. In the gill no distinct change in the amino acid pool occurred during 14 days of exposure, with the exception of a decrease in serine. When transferred again to 38%. S, a strong overcompensation occurred in alanine, proline, glycine and serine, and a smaller in glutamate and threonine.
• 5.5. No evidence for anaerobic metabolism in the decrease of the amino acid pool was found.
• 6.6. M. galloprovincialis is less able to adapt to low salinities than the more euryhaline M. edulis.

     

Driving forces for the uphill transport of amino acids into epidermal brush border membrane vesicles of the sea anemone,Anemonia sulcata (Cnidaria,anthozoa)

• 1.1. The transport of amino acids into membrane vesicles prepared from epidermal tentacle tissue of the sea anemone, Anemonia sulcata, depends on an electrochemical potential difference caused, e.g. by sodium chloride gradients.
• 2.2. Potassium or choline chloride gradients energized the transport less effectively than sodium chloride gradients. Both Na⁺-ions and Cl⁻-ions were required for the amino acid transport.
• 3.3. The uphill transport of amino acids along the downhill movement of driver ions (sodium chloride gradient conditions) was characterized by an overshoot; under sodium chloride equilibrium conditions, however, an accumulation of amino acids within the vesicles could not be measured.
• 4.4. Potassium diffusion potentials in combination with valinomycin indicated that hyperpolarization (vesicle inside negative) and hypopolarization (vesicle inside positive) enhanced or depressed the accumulation of amino acids within the vesicles.
• 5.5. Being at the phylogenetic base of the Eumetazoa, cnidarians show characteristics for the transmembrane transport of amino acids comparable to those established for vertebrates.

     

Metabolic responses of the mussels Perna viridis and Perna indica to declining oxygen tension at different salinities

• 1.1. Oxygen uptake and ammonia loss were monitored during responses to reductions of both salinity and oxygen tension (PO₂) in the marine mussels Perna viridis and Perna indica from southern India.
• 2.2. The proportional contribution of protein to total catabolic substrates under natural environmental conditions was as much as 96% in P. viridis, relative to only 19% in P. indica.
• 3.3. Normoxic oxygen consumption remained statistically unchanged in P. viridis conditioned to salinities between 32 and 15‰, with no obvious signs of distress. Although equally unaffected at salinities between 32 and 20‰, P. indica showed significantly reduced oxygen uptake following transfer from 32 to 15‰, and had died within the next 7 days.
• 4.4. At salinities greater than 20‰, P. viridis was better able than P. indica to regulate oxygen consumption independent of PO₂.
• 5.5. P. indica showed a compensatory increase in oxyregulatory capacity at 15‰. This exceeded unstressed abilities, helping to maintain albeit reduced oxygen uptake throughout wider ranges of PO₂.
• 6.6. Different responses recorded in each of these tropical and often intertidal species were in accordance with their natural distributions. Nevertheless, the oxyregulatory capacity in both species was higher than in bivalves from temperate and/or subtidally restricted habitats.

     

Changes in free amino acid composition in haemolymph of larvae of the wax moth,Galleria mellon ella L., during cold acclimation

• 1.1. Free amino acids were analysed in the haemolymph of Galleria mellonella larvae by HPLC chromatography with o-phthaldialdehyde (OPA)-l-thio-β-d-glucose as derivatization agent.
• 2.2. Fourteen primary amino acids were detected among which glutamine, alanine, γ-aminobutyric acid (GABA) and glycine predominated and constituted 67.7% of the amino acids found.
• 3.3. The concentration of GABA increased significantly with the age of larvae entering the wandering phase and reached a maximum during metamorphosis.
• 4.4. Analysis of cold-acclimated larvae revealed a net increase of free primary amino acids from 96 to 151.8 μmol/ml during consecutive acclimation to 0°C within 20 days and to 205.4μmol/ml during cold shock injury at 0°C (3 hr).
• 5.5. The bulk of this increase was accounted for by alanine, glycine, phenylalanine and lysine.

     

Effect of eyestalk ablation on oxygen consumption of callinectes similis exposed to salinity changes

• 1.1. The effect of eyestalk ablation on preadults of Callinectes similis exposed to a constant salinity (30%.) and to simulated tidal changes in salinity (30-11 to 30%.) were measured.
• 2.2. In constant salinity, crabs showed a persistent respiratory rhythm, with a maximum oxygen consumption during the day. Under these conditions, ablation significantly increased the respiratory rate but not the rhythm.
• 3.3. In variable salinities, the highest respiratory rates occurred in salinities of 11 and 16%. during the night. In these crabs, ablation of eyestalks and subsequent injection of eyestalk extracts did not alter the respiration rate rhythm.
• 4.4. The circadian rhythm is controlled by the periodicity of environmental changes instead of the influence of eyestalk hormones.
• 5.5. Regulation of metabolism in C. similis associated with osmoregulation involves other neurosecretory organs.

     

Changes in composition and proteolytic enzyme activities of artemia during early development

• 1.1. The proximate composition, total and free amino acids, and proteases of Artemia nauplii were determined during early development.
• 2.2. Moisture increased from 71.0% to 80.8%, crude protein decreased from 13.2% to 8.8%, crude fat and ash varied slightly.
• 3.3. The total amino acids decreased. Free amino acids changed in three patterns.
• 4.4. Trypsin, chymotrypsin, carboxypeptidase A, B and cathepsin B and C increased in activity. The activity of trypsin was lower, while cathepsin B and C were the highest.
• 5.5. The protease activities were maximal at pH 7.5 and 8.0, and at 45°C on casein.
• 6.6. The optimal pH for carboxypeptidase A was 4.0, for carboxypeptidase B was 4.5, for trypsin and chymotrypsin were 7.0–7.5. The protease(s) active at pH 9.0–9.5 were to be determined.

     

Chemical feeding stimulants for the herbivorous fish,Tilapia zillii

• 1.1. Chemical feeding stimulants for an herbivorous fish, Tilapia zillii have been determined by fractionation and bioassay of substances derived from a model food plant.
• 2.2. Stimulation was produced by amino acids; glutamic acid, aspartic acid, serine, lysine and alanine produced the bulk of stimulatory activity.
• 3.3. These amino acids are among the most abundant in the test plant, and are markedly different from the amino acids found to stimulate feeding in carnivorous fish.
• 4.4. On the basis of these results, a chemically-mediated mechanism of feeding niche separation is postulated.

     

A starvation test to determine optimal salinities for larval freshwater prawns,Macrobrachium rosenbergii

• 1.1. A starvation test was conducted in small beakers with stage 1 (S1) and stage 2 (S2) Macrobrachium rosenbergii larvae to determine optimal salinities.
• 2.2. Experiments were first performed with S2 larvae at 13 ppt to identify a suitable medium made with artificial sea salts.
• 3.3. A broad-range (0–35 ppt) and a subsequent narrow-range (9–16 ppt) salinity experiment with S2 larvae were used to identify 13 ppt as the optimal salinity, with 12 ppt as the next best; this agrees well with most previous estimates of optimal salinities for rearing larvae.
• 4.4. S1 larvae were also tested in a narrow-range salinity experiment but were not used further because, unlike starved S2 larvae, they molted during the experiment.
• 5.5. Identification of the optimal salinity was not affected by 50% daily water exchange or by bright light.
• 6.6. Exposure of larvae to three different salinities—7, 13 and 19 ppt—during S1 influenced the width of the optimal salinity range for S2 larvae.

     

The free amino acids of muscle in the cephalochordate Branchiostoma,the cyclostome Myxine and some fishes

• 1.1. Branchiostoma and Myxine have the highest concentrations of amino acids (207 and 234 mM) of the five species investigated.
• 2.2. The predominant amino acids are glycine, proline, alanine, taurine, serine and valine, which form 83–98% of the total, except in Latimeria (60%).
• 3.3. Total amino acids are considered from the point of view of osmotic concentration in relation to other nitrogenous compounds of muscle.

     

Specific dynamic action (SDA) in the supralittoral isopod,Ligia pallasii: Identification of components of apparent SDA and effects of dietary amino acid quality and content on SDA

• 1.1. Specific Dynamic Action (SDA) effects of diet were investigated in the supralittoral isopod, Ligia pallasii, using defined chemical diets.
• 2.2. “Apparent SDA”, or the total rise in metabolic rate following a meal, was resolved in animals eating a nutritionally complete chemical diet into three components: 8% mechanical costs of moving food through the gut, 40% “excitement costs” due to investigator disturbance and presence of food, and 52% SDA.
• 3.3. Excitement costs in animals exposed to food but which chose not to eat showed non-significant variation between diets containing different levels of chemical nutrients, but were significantly less on a diet containing only cellulose and agar.
• 4.4. SDA increased with increasing concentration of amino acids in the diet.
• 5.5. Substitution of whole-protein casein for free amino acids in the diet had no significant SDA effect, while substitution of free amino acids in the ratio found in casein more than doubled the SDA effect.
• 6.6. Deletion of alanine from the diet caused no significant effect on SDA, while deletion of phenylalanine caused a highly significant elevation in SDA.

     

Amino acid synthesis during hyperosmotic stress in penaeus aztecus postlarvae

• 1.1. Glycine, proline, and taurine are the quantitatively most important amino acid osmolytes in Penaeus aztecus postlarvae.
• 2.2. Taurine dominates the amino acid pool in low salinity, while proline dominates the amino acid pool at higher salinities.
• 3.3. Although not major contributors to the pool, glutamate and alanine are constitutively synthesized from [¹⁴C]glucose and [¹⁴C]glutamate under constant salinity and under hyperosmotic stress treatments.
• 4.4. Proline synthesis from [¹⁴C]-precursors is apparent under constant high (but not low) salinity and is significantly induced by hyperosmotic stress.
• 5.5. No appreciable glycine synthesis was observed from [¹⁴C]glucose or [¹⁴C]glutamate under any experimental conditions.

     

Activities of the three ammonia-forming enzymes in the tissues of the brackish-water bivalve Corbicula japonica

• 1.1. Activities of the three ammonia-forming enzymes, glutamate dehydrogenase, AMP deaminase and serine dehydrase (SerDH), were measured in tissues of gill, digestive diverticula, mantle and foot muscle of the brackish-water bivalve Corbicula japonica.
• 2.2. High levels of SerDH activity were detected in gill and digestive diverticula, while the activity levels of the other two enzymes were low.
• 3.3. The result suggests the significance of SerDH in amino acid degradation of this species.

     

Stomach digestion and its effect upon protein hydrolysis in the intestine of rainbow trout (Salmo gairdneri Richardson)

• 1.1. The in vitro digestion of soya protein by pancreatic proteases of the trout was measured following various conditions of stomach digestion.
• 2.2. Peptic hydrolysis results in an increase of small peptides.
• 3.3. Acid treatment and peptic proteolysis do not affect the degradation of insoluble proteins or the formation of free amino acids during intestinal digestion, but they do lead to a significant shift from soluble polypeptides to di- and oligopeptides.

     

Specific dynamic action (SDA) in the supralittoral isopod,Ligia pallasii: Effect of ration and body size on SDA

• 1.1. Ration and body size effects on specific dynamic action (SDA) were investigated in the supralittoral isopod Ligia pallasii using seaweed and chemical diets.
• 2.2. SDA increased asymptotically with ingested meal size for all diets.
• 3.3. Body weight had a significant positive effect on SDA for only one of the six diets tested, but weak tendencies were present in the data for the other diets.
• 4.4. SDA appeared to increase geometrically with increasing concentration of amino acids at high ration levels.

     

Chemical composition of North Atlantic krill

• 1.1. The main chemical components of Meganyctiphanes norvegica (M. Sars), Thysanoessa inermis (Krøyer) and T. raschii (M. Sars) have been examined.
• 2.2. Protein accounted for 42–47% of the dry weight of M. norvegica and 32–50% of the dry weight of the Thysanoessa species. On a wet weight basis, the protein content was relatively constant and independent of season.
• 3.3. The dominating amino acids in the bulk protein of the krill were glutamic acid/glutamine, aspartic acid/asparagine, glycine, alanine, lysine and leucine.
• 4.4. Lipids were present in amounts of 13–29% of the dry weight in M. norvegica, 15–50% in T. inermis and 12–44% in T. raschii, and the lipid content varied with season.
• 5.5. The main nitrogen extractives in krill, expressed on a dry weight basis, were free amino acids (5–10%), trimethylamine oxide (about 4%), peptides (about 1%) and nucleotides (0.4–1.3%). Trimethylamine and ammonia were present in very low concentrations in living krill.
• 6.6. The amino acids taurine, glycine, proline, arginine, sarcosine and alanine made up 89–93 mol% of the free amino acid pool.
• 7.7. The ash content of krill was in the order of 10–13% of the dry weight, and fluoride represented 1040 and 3200 ppm in the Thysanoessa species and M. norvegioca, respectively.

     

Role of arginase transferred from the vesicula seminalis during mating and changes in amino acid pools of the spermatophore after ejaculation in the silkworm,Bombyx mori

.

• 1.1. Ejaculation of seminal fluid into the spermatophore formed inside the female bursa copulatrix terminates 20 min after the beginning of copulation of Bombyx mori. The amounts of amino acids transferred are small, but the amounts of amino acids in the spermatophore continue to increase after this time due to protein degradation and amino acid interconversions.
• 2.2. Arginase, which has high activity in the vesicula seminalis, is transferred to the spermatophore without loss of activity during ejaculation. During mating, ornithine and much urea are formed in the spermatophore, indicating activity of the transferred arginase.
• 3.3. In the spermatophore, marked increase of alanine with low concentrations of ornithine and glutamate suggests the presence of a pathway for the active formation of 2-oxoglutarate with pyruvate via glutamate from arginine. During mating, proline and glutamine also increase, but at low rates.
• 4.4. Of the exocrine glands in the male reproductive system, the vesicula seminalis secretes the highest concentration of glutamate (30% of the total amino acids); serine is the amino acid present at highest concentration in secretions of other glands (20–30%). No urea was found in the secretions of any of the glands.

     

Thermochemistry of N-C bonds hydrolysis in amides,peptides, N-acetyl amino acids and high-energy N-C bonds hydrolysis in N-acetyl imidazole and urea

• 1.1. The reaction enthalpies of hydrolysis of amides, peptides and N-acetyl amino acids were calculated for both ionized and un-ionized forms of reaction components.
• 2.2. The average values of reaction enthalpies of amides, peptides and N-acetyl amino acids hydrolysis were essentially different from each other for ionized forms of reaction components and were equal for un-ionized forms of reaction components in the error interval.
• 3.3. As an example of high-energy N-C bonds N-acetyl imidazol and urea were discussed. It was found that the reaction enthalpies of hydrolysis of above compounds were different from analogous thermodynamic values of hydrolysis of amides, peptides and N-acetyl amino acids for any forms of components.

     

Effect of dietary d-amino acids on growth,survival, ammonia excretion and specific dynamic action in the supralittoral isopod,Ligia pallasii

• 1.1. Growth, survival, ammonia excretion and Specific Dynamic Action (SDA) were assessed in the supralittoral isopod Ligia pallasii eating chemical diets with differing proportions of d- and l-amino acids. Growth and survival decreased in direct proportion to increasing dietary intake of d-amino acids.
• 2.2. Survival on diets with greater than 50% content of d-amino acids (based on total amino acids in diet) was lower than that expected based on previous work, suggesting a deleterious effect of the d-isomers.
• 3.3. Ammonia excretion and SDA correlated negatively with increasing dietary content of d-amino acids.
• 4.4. The general conclusion is that d-amino acids play no role in anabolic or energy metabolism in Ligia, and that poor performance at higher dietary levels of d-amino acids may relate to their interference with transport pathways for the normal l-forms.

     

Proteolysis post mortem in North Atlantic krill

• 1.1. The autoproteolytic processes in selected species of North Atlantic krill, Meganyctiphanes norvegica (M. Sars), Thysanoessa inermis (Krøyer) and T. raschii (M. Sars) have been examined at 0°C by following the release of peptides and free amino acids.
• 2.2. The krill contains high levels of peptide hydrolases, and autoproteolysis seems to be due mainly to digestive enzymes localized in the hepatopancreas and the intestinal tract of the animals.
• 3.3. During autoproteolysis the individual amino acids were generally released at rates corresponding to their proportion in the bulk protein of the krill. The major exceptions were alanine which accumulated in amounts larger than was to be expected from the composition of the krill protein, and glutamic acid/glutamine, aspartic acid/asparagine, arginine, and to some extent glycine, proline and serine, which accumulated to a lesser extent than was to be expected.
• 4.4. Storage of krill for 1 week resulted in only minor changes in the total content of amino acids as determined after acid hydrolysis, with the exception of alanine which increased in concentration.
• 5.5. The results suggest that the formation of free alanine is partly due to reactions other than proteolysis.
• 6.6. The release of free amino acids was accompanied by a considerable increase in the amount of small peptides, and glutamic acid/glutamine, aspartic acid/asparagine, glycine and proline tended to accumulate in these peptides.
• 7.7. The autoproteolytic activity of the Thysanoessa species showed seasonal variations, probably in response to food availability. In the case M. norvegica, the results suggest that there are smaller fluctuations in the level of proteolytic enzymes, probably indicating less pronounced variations in the food intake over the year.