Applications of Hydrolytic and Decarboxylating Enzymes in Biotransformations

Peptides, and oligosaccharides and glycosides, can be synthesised by making use of the ‘reverse hydrolytic activity’ of proteases and glycosidases respectively. In applying these enzymes to the practical synthesis of these classes of compound, several factors need to be considered, namely the need to shift the rate-determining step through the use of activated substrates, the need to minimise competing hydrolysis of these and the need to minimise hydrolysis of the products. In spite of these problems, the enzymatic methods have many attractive features, not least amongst which is the absolute control of stereochemistry in acyl transfer and glycosyl transfer respectively.     

Regional environments, life-history patterns, and habitat use of spirostreptid millipedes in arid regions

Among diplopods with desert populations, only three species of Spirostreptida have been studied in an ecological context. The present review compares regional environments, life-history patterns, and uses of habitat by Orlhoporus ornatus (Girard) from southwestern North America, Archispirostreptus tumuliporus judaicus (Attems) from the eastern Mediterranean seaboard, and Harpagophora nigra (Attems) from southwestern Africa. Published and unpublished studies are used to explore evidence for convergence among these species, as opposed to traits adapting them to physical aspects of given regions or habitats. Unlike A. t. judaicus, O. ornatus and H. nigra are relatively restricted to arid habitats, although populations of all three species experience a variety of rainfall regimes and regional topographies. Where studied, O. ornatus and H. nigra hibernate during the long, often cool or cold dry season; they forage following warm-season rains. A. t. judaicus , in contrast, forages during its long, warm dry season and hibernates in the cool, wet winter. Populations from the Judaean and Negev deserts differ from those inhabiting a mesic habitat (Megiddo) closer to the coast in regard to rates of development, seasonal activity and seasonal water balance. Convergence in the form of well-developed desiccation resistance characterizes the two strictly desert species. All three species, together with other subtropical millipedes exposed to long dry seasons, are convergent with respect to patterns of diel surface activity and use of shelter. However species- and habitat-specific life-history features such as the seasonal timing of dormancy and emergence tend to mask convergence at the habitat level. Hence, the independent evolution of the three species with desert populations has resulted in life histories and habitat use that combine a moderate amount of convergence with considerable opportunistic adaptation to regional and local conditions.     

Determination of taxonomic resolution capacity of conventional one-dimensional SDS-polyacrylamide gel electrophoresis of whole-cell proteins using Enterobacteriaceae

The capacity of one-dimensional SDS-PAGE of whole bacterial cells to both separate and cluster taxonomic units is studied using members of Enterobacteriaceae as test material. The results show that intraspecies variation can be detected and on the other hand the degree of taxonomic divergence which still can be grouped together is determined. In addition the system has high tolerance to changes in cell culture conditions making the usage of SDS-PAGE suitable for applications where rapid and reliable bacterial identification is needed.     

Releaser and primer pheromones in Collembola

In Collembola, pheromones appear to be present in the faecal pellets. Pheromone release after cessation of faeces production points to the digestive tract as a possible site of biosynthesis.During the pre-moulting periods Collembola do not react to pheromones, possibly due to their low activity at that time, whereas the production of the pheromones continues.Starvation periods of up to 14 days diminish pheromone release but do not cause complete cessation. Production per animal seems to decrease at increasing densities.The effect of pheromones on the reproductive efficiency of Collembola is discussed in the context of their physiological and behavioural ecology.     

DIGESTION SURVIVAL OF ALGAE: SOME ECOLOGICAL COMPARISIONS BETWEEN FREE SPORES AND PROPAGULES IN FECAL PELLETS1

Benthic algae survive passage through the digestive tract of different types of grazers, either regenerating new tissues or releasing protoplasts and swarmers which act as accessory means of reproduction to generate new individuals. The ecological importance of this phenomenon is evaluated here by studying spores and fecal pellets. Alga fragments egested alive inside fecal pellets have some ecological advantages over free propagules. In intertidal habitats, the sticky nature of the pellets permits attachment to the substratum while protecting the contained algae from desiccation. In subtidal habitats, the pellets sink 8–22 times faster than the fastest sinking algal propagule. Quantification of swarmers and protoplasts released from algal fragments in the fecal pellets indicates densities of about 217 germlings·cm^?2 and values of 300–700 propagules per pellet. Extrapolation of these results to field conditions based on number of grazers and pellets suggest production figures varying from 0.5–3.5 × 10⁵to 1.3–1.5 × 10⁶ propagules·m^?2·12 h^?1. These values are within known density ranges of settling spores in the field. The ecological significance of the phenomenon may be especially important at seasons when the density of grazers increases and the normal reproductive activity of the algae being consumed decreases.     

Genetic similarity in long-eared owl communal winter roosts: a DNA fingerprinting study

The long-eared owl is a nocturnal predator which winters communally and breeds in the same areas in loose colonies during the spring. We tested the hypothesis that roosts, particularly stable roosts, are formed by close relatives, a condition under which group-related behaviours such as information sharing or helping at nest are more likely to be developed. DNA fingerprinting analysis was used to examine genetic similarity within and between two long-eared owl populations, one wintering in a traditional roost and the other in an unstable roost, and both breeding around their roosting sites. Although genetic similarity within roosts was higher than that between roosts, the difference was not significant. Observed genetic similarity within roosts was smaller than that reported in the bird species whose roosts work as information centres. On the other hand, the presence of some closely related individuals in the roost and behavioural observations suggest that co-operation between kin might have occurred, at least in one of the two study sites.     

Influence of culture parameters on lignin metabolism byPhanerochaete chrysosporium

Culture parameters influencing metabolism of synthetic¹⁴C-lignins to¹⁴CO₂ in defined media have been studied in shallow batch cultures of the ligninolytic wood-destroying HymenomycetePhanerochaete chrysosporium Burds. Study of the effect of O₂ concentration in the gas phase above non-agitated cultures indicated essentially complete absence of attack on the lignin polymer at 5% O₂ in N₂, and a 2- to 3-fold enhancement by 100% O₂ as compared to air (21% O₂). Agitation of the cultures resulting in the formation of mycelial pellets greatly suppressed lignin decomposition. The optimum culture pH for lignin decomposition was 4 to 4.5, with marked suppression above 5.5 and below 3.5. The source of nutrient nitrogen (NO ₃ ^– , NH ₄ ⁺ , amino acids) had little influence on lignin decomposition, but the concentration of nitrogen was critical; decomposition at 24 mM was only 25–35% of that at 2.4 mM N. Thiamine was the only vitamin required for growth and lignin decomposition. Under the optimum conditions developed, decomposition of 5 mg of synthetic lignin was accompanied by utilization of approximately 100 mg of glucose. The influence of the various culture parameters was analogous for metabolism of synthetic lignin labeled in the ring-,side chain-, and methoxyl carbon atoms.     

Biocatalytic synthesis of disaccharide high-intensity sweeterner sucralose via a tetrachlororaffinose intermediate

A second high-yielding bioorganic synthesis of the highintensity sweetener sucralose (4,1',6'-trichloro-4,1',6'-trideoxygalactosucrose) is described. This procedure involves the chemical chlorination of raffinose to form a novel tetrachloroaffinose intermediate (6,4',1',6'-tetrachloro-6,4',1',6'-tetradeoxygalactoraffinose; TCR) followed by the enzymic hydrolysis of the alpha-1-6 glycosidic bond of TCR to give sucralose and 6-chlorogalactose. Commercial enzyme preparations and microorganisms were screened to select alpha-galactosidases which have high catalytic activity on this compound. The most active enzyme was produced by a strain of Mortierella vinacea and had a maximum rate of 118 mumol sucralose/g dry weight cells/h, which was approximately 5% of the activity toward raffinose, and a K(m) of 5.8 mM toward TCR. The enzyme could be used in the form of mycelial pellets in a continuous packed bed column reactor. The reaction was also studied in a water-immiscible hydrophilic organic solvent, such as methyl isobutyl ketone, to overcome the poor aqueous solubility of TCR and to increase volumetric productivity. Synthesis of raffinose was achieved from saturated aqueous solutions of galactose and sucrose using a selected alpha-galactosidase from Aspergillus niger. When raffinose is used as a starting material for sucralose synthesis, this route has fewer steps than either the preceeding method using glucose-6-acetate as an intermediate or the complete chemical synthesis from sucrose. The relative merits of the two bioorganic routes and the utility of such methods to synthesize new sugars are discussed.