Adaptation of membrane fluidity of rye wheat seedlings according to temperature

Order parameters of chloroplast membrane lipids of rye wheat seedlings differing in cold hardiness were compared before after hardening. Seedlings grown at 25° exhibited similar membrane microviscosities. When hardened, the cultivars most resistant to freezing temperatures possessed the most fluid membranes, while those sensitive to cold were unable to alter them. Changes in linolenic acid levels alone cannot be responsible for the observed phenomena.     

Adaptation ofDrosophila enzymes to temperature. III. Evolutionary conservatism in mitochondrial enzymes

Summary The evolutionary behavior of two mitochondrial enzymes (L-glycerol 3-phosphate:cytochrome c oxidoreductase E.C.1.1.1.95,GPO, and L-malate: NAD⁺ oxidoreductase, E.C.1.1.1.37, m-MDH) obtained from several temperate and tropicalDrosophila species was examined by comparing their catalytic properties, which related to temperature (K_m-E_a-Q₁₀-Thermostability). MitochondrialGPO or m-MDH obtained either from temperate or from tropical species was found to exhibit similar catalytic properties while for both cytosolic enzymes, theGPDH and s-MDH, K_m patterns were similar among species from the same thermal habitat and different between thermal habitats. In combination with other observations reported in the literature these facts support the view that the function, and probably the structure, of mitochondrial enzymes are better conserved in evolution than those of the corresponding enzymes found in the cytosol. It is proposed that the relative invariance of the mitochondrial enzymes structure is probably linked to a necessary relative invariance of molecular interactions inside the mitochondrion.     

Adaptation of Drosophila enzymes to temperature--I. Acetylcholinesterase and NADP-dependent isocitrate-dehydrogenase.

1. The relationship between Km and assay temperature was examined in three tropical and two temperate Drosophila species, and in the cosmopolitan species, D. melanogaster, for isocitrate dehydrogenase and acetylcholinesterase. 2. For both enzymes Km patterns were similar among species from the same habitat, and different between habitats. No such parallelism was seen with respect to thermal inactivation. 3. The Q10 values in general reflected temperature dependent changes in Km, but exceptions were noted.     

A note on stability of purified enzymes to dehydration

Highly purified enzymes from commercial sources were dried from aqueous solutions over CaCl₂ or CaSO₄ at 25° c, 4° c or −4° C. Several heat stable enzymes containing SS groups (chymotrypsin, lipase, pepsin and trypsin) retained most of their activity following drying while papain and ribonuclease retained about one-fourth of their original activity. Most SH containing enzymes lost most if not all activity during drying (catalase, hexokinase, glucose and xanthine oxidases, alcohol, glutamate, isocitrate and glucose-6-phosphate dehydrogenases). Lactic dehydrogenase was the exception in this group retaining 75% of its original activity after drying. Neither sucrose nor mannitol were effective in protecting ribonuclease against inactivation during drying. Temperature during drying had little effect on inactivation.     

Adaptation to larval crowding in Drosophila ananassae leads to the evolution of population stability

Density-dependent selection is expected to lead to population stability, especially if r and K tradeoff. Yet, there is no empirical evidence of adaptation to crowding leading to the evolution of stability. We show that populations of Drosophila ananassae selected for adaptation to larval crowding have higher K and lower r, and evolve greater stability than controls. We also show that increased population growth rates at high density can enhance stability, even in the absence of a decrease in r, by ensuring that the crowding adapted populations do not fall to very low sizes. We discuss our results in the context of traits known to have diverged between the selected and control populations, and compare our results with previous work on the evolution of stability in D. melanogaster. Overall, our results suggest that density-dependent selection may be an important factor promoting the evolution of relatively stable dynamics in natural populations.     

Adaptation of enzymes to temperature: myofibrillar adenosine triphosphatase from two terrestrial isopod species

A comparison of the temperature profiles of the myofibrillar ATPase in Porcellio spinicornis and Metoponorthus pruinosus showed that Porcellio enzyme had a maximum activity between pH 6.8-7.2 at 9 degrees C, and that from Metoponorthus between pH 7.0-7.4 at 45 degrees C. The energy of activation for Porcellio enzyme was estimated to be 2.964 kcal/mol and that for Metoponorthus enzyme 9.91 X 10(-1) kcal/mol. The significance of these findings in relation to the natural habitats of these two species is discussed.     

Adaptation of Drosophila enzymes to temperature IV. Natural selection at the alcohol-dehydrogenase locus

Studies were carried out on the temperature-dependent kinetic properties (K_m, Q₁₀, E_a, thermostability) of alcohol-dehydrogenase allozymes from D. melanogaster. It was shown that there is a parallelism between the biochemical properties of the enzymes and the behaviour of the genes in natural and cage populations. Furthermore, the relationship between the temperature-dependent kinetic properties of alcohol dehydrogenase and assay temperature was examined in three tropical and two temperate Drosophila species. K_m patterns were similar among species from the same habitat and different between habitats. No such parallelism was seen with respect to thermal inactivation. The Q₁₀ values in general reflected temperature-dependent changes in K_m. It is discussed that the mechanistic connection observed between the biochemical and population levels (intra-and interspecifically) strongly suggests that temperature acts as a selective factor on the structural Adh locus in the genus Drosophila.     

Protein engineering to change thermal stability for food enzymes.

In this review we have briefly indicated how the present state of knowledge allows proteins to be mutated to increase or decrease stability. We have discussed experiments on both model proteins and those of relevance to the food industry, and show how hydrophobic forces are a major driving force for folding as well as having a major role in thermostability. We have also indicated the large contribution that hydrogen bonding, electrostatic interactions and, in a less well predicted way, disulphide bridges make to thermostability.     

A new method to evaluate the thermal stability of lyophilized enzymes

Summary The thermal inactivation of lyophilized chymotrypsin was studied at controlled water activities. At 60 °C the enzyme showed good stability except at aw 0.97, whereas at 75 °C considerable inactivation occured at most water activities. Increasing the amount of buffer on the preparation decreased the stability significantly. The optimal temperature of enzymatic activity was increased 14 °C, when the water activity was decreased from 1 to 0.5.     

Adaptation of intestinal cell membrane enzymes to low temperatures in the Antarctic teleost Pagothenia bernacchii

The enzymatic activity (expressed as milliunits per milligram total proteins) of three intestinal brush-border membrane enzymes, leucine aminopeptidase, alkaline phosphatase and maltase, measured over a range of temperatures between 1.5 and 37 °C, has been found to be much higher in the Antarctic fish Pagothenia bernacchii than in the temperate fish Anguilla anguilla. To explain this experimental observation the apparent Michaelis-Menten constant, the maximal velocity, the activation energy values and the thermal stability of these three enzymes were measured. The apparent Michaelis-Menten constant values of leucine amino peptidase and alkaline phosphatase were different in the intestine mucosal homogenate of the two fish at each measured temperature (from a minimum of 2.5 to a maximum of 37 °C). However, the values found at 2.5 °C for the Antarctic species and 15 °C for the eel where comparable. Furthermore, its value was unchanged in eel intestine apical membranes, both in the presence and without enzyme lipid microenvironment. While the maximal enzymatic activities of the leucine aminopeptidase and maltase did not decrease without their enzyme lipid microenvironment, produced by treatment with Triton X-100, the impairment of alkaline phosphatase maximal activity cannot be significantly differentiated from a non-specific inhibitory effect of the detergent. The activation energy values of leucine amino peptidase, alkaline phosphatase and maltase were lower in the Antarctic fish (11.7, 5.6 and 11.8 kcal·mol^-1, respectively) than in the eel (13.6, 7.6 and 13.1 kcal·mol^-1, respectively). The thermal stability of alkaline phosphatase and maltase is different in Pagothenia bernacchii and Anguilla anguilla intestinal homogenate.Abbreviations BBM brush border membrane - E _a activation energy - EGTA ethyleneglycol-bis-(-amino ethylether)N, N-tetraacetic acid - HEPES 2-[4-(2-hydroxyethyl)-1-piperazinyl]-ethane sulphonic acid - Km_app apparent Michaelis-Menten constant - PMSF phenylmethyl-sulphonyl fluoride - TRIS TRIS (hydroxymethyl)-aminomethane     

Aquatic snails of the Bulinus africanus group in Zambia identified according to morphometry and enzymes

The Bulinus africanus species group (Planorbidae) of freshwater snails has been reported to be represented in Zambia by two species, B. africanus (Krauss) and B. globosus (Morelet), both named as intermediate hosts for Schistosoma haematobium. Uncertainty in identification of these snails from morphology led to the present investigation, combining morphometry (shell and copulatory organ) with enzyme analysis. Observations of both kinds were made usually on the same individual snails, from collecting sites mostly in the Lusaka area or at Lake Kariba. Particular attention was given to the proportional relationship between the penis sheath and the preputium of the copulatory organ, a character used previously to distinguish B. africanus from B. globosus in south-eastern Africa. The enzyme profile MDH-1, AcP-2, PGD-1 and PGM-2 was common to all snails examined from 25 populations; GPI and HBDH were polymorphic. The enzyme data indicate that the samples represent a single species. Shell characters varied continuously. The copulatory organ was generally of the form known for B. globosus. Although the copulatory organ of a few individuals had proportions overlapping the range reported for B. africanus, the present variation was continuous and was not bimodal. It is concluded that all these specimens are conspecific and may be identified as B. globosus. Previous identifications of B. africanus from Zambia appear to need substantiation and it seems that if this species is present at all in the sampled areas, it must be uncommon. It is relevant in regard to possible strain differences within S. haematobium in Zambia, that our observations indicate that only a single species of intermediate host is involved in transmission.     

Thermal and chemical stability of solid-state enzymes

The thermal deactivation of solid-state acid-phosphatase has been studied, in the presence and in the absence of organic solvents. The experimental results have been modelled in terms of a single-step, non-linear, irreversible kinetic model. Enzyme stability was strongly affected by deactivation temperature and initial water content of the enzyme preparation. In contrast, no direct influence of solvent hydrophobicity was detected. The results were compared with those obtained in aqueous solution.