Knowledge-based modeling of the D-lactate dehydrogenase three-dimensional structure

A three-dimensional structure of the NAD-dependent D -lactate dehydrogenase of Lactobacillus bulgaricus is modeled using the structure of the formate dehydrogenase of Pseudomonas sp. as template. Both sequences share only 22% of identical residues. Regions for knowledge-based modeling are defined from the structurally conserved regions predicted by multiple alignment of a set of related protein sequences with low homology. The model of the D -LDH subunit shows, as for the formate dehydrogenase, an α/β structure, with a catalytic domain and a coenzyme binding domain. It points out the catalytic histidine (His-296) and supports the hypothetical catalytic mechanism. It also suggests that the other residues involved in the active site are Arg-235, possibly involved in the binding of the carboxyl group of the pyruvate, and Phe-299, a candidate for stabilizing the methyl group of the substrate. © 1995 Wiley-Liss, Inc.     

Nitrogen-fixing cyanobacteria as source of phycobiliprotein pigments. Composition and growth performance of ten filamentous heterocystous strains

Ten strains of filamentous, heterocystous nitrogen-fixing blue-green algae (cyanobacteria) were screened for growth performance and tolerance to temperature, pH, irradiance and salinity, together with their potential as producers of phycobiliprotein pigments. Phycobiliproteins typically accounted for about 50% total cell protein, the prevalent type being C-phycocyanin, followed by alloppycocyanin, with levels of 17 and 11% d.wt, respectively, in some strains of Anabaena and Nostoc. C-phycoerythrin was the major pigment in several Nostoc strains, reaching 10% d.wt. Some strains represent, therefore, excellent sources of one or more phycobiliproteins. All strains tolerated an irradiance of ca 2000 μmol photon m^-2 s^-1. Anabaena sp. ATCC 33047 and Nostoc sp. (Albufera) exhibited the widest optimum range of both temperature (30–45 and 25–40 °C) and pH (6.5–9.5 and 6.0–9.0) for growth, the former also showing significant salt tolerance. In an outdoor open system, productivity of cultures of two phycoerythrin-rich strains of Nostoc was over 20 g (d.wt) m^-2 d^-1 during summer. The growth performance of the allophycocyanin-rich Anabaena sp. ATCC 33047 in outdoor semi-continuous culture has been assessed throughout the year. Productivity values under optimized conditions ranged from 9 (winter) to 24 (summer) g (d.wt) m^-2 d^-1.     

In vitro pollen germination of species and two interspecific hybrids from the genus Brassica

In vitro pollen germination of five species and two interspecific hybrids from the genus Brassica was tested in four media. Genetically fixed differences in the demands for optimal pollen germination among species were found. The experiments were designed to define optimal content of mineral salts, sugar, and PEG for every investigated species or hybrids. The differences found among species are discussed in relation to the evolutionary trend.     

Constrained C-terminal hexapeptide neurotensin analogues containing a 3-oxoindolizidine skeleton

Summary In order to enforce different spatial orientations in the C-terminal hexapeptide of neurotensin (NT_8–13) and to gain information about the importance of the 10–11 peptide bond for binding to NT receptors, the Pro¹⁰-Tyr¹¹ fragment has been replaced with (2R,8S,8aR)-, (2S,8S,8aR)-, (2S,8S,8aS)-, (2S,8R,8aS)- and (2R,8R,8aS)-8-amino-2-benzyl-3-oxoindolizidine-2-carboxylic acid. Molecular dynamics calculations and energy minimization studies have shown that, contrarily to the Pro-Tyr moiety, none of these indolizidines display a tendency to adopt type I and III -turns, but those having (8S,8aR) or (8R,8aS) stereochemistry essentially adopt extended conformations and the (8S,8aS) stereoisomer prefers a nonstandard folding. The four diastereomeric NT_8–13 analogues incorporating (8S,8aR) or (8R,8aS) indolizidines displayed binding affinities for the brain NT receptor similar to that of [Ala¹¹]-NT_8–13 and only five- to ninefold lower than that of the corresponding analogue, [Phe¹¹]NT_8–13. Although this slight decrease could be attributed to differences in conformational behavior between these constrained NT_8–13 analogues and [Phe¹¹]NT_8–13 or NT_8–13, it is not clear whether the -turn around Pro¹⁰-AA¹¹ (AA=Phe, Tyr) is conserved upon receptor binding. An excessive restriction in the motions of the aromatic side chain, imposed by the highly steric constraint of the indolizidine moiety, emerges as an alternative explanation. The findings reported here demonstrate the possibility of replacing the Pro¹⁰-Tyr¹¹ dipeptide in NT_8–13 with a non-peptide residue without affecting considerably the affinity for brain NT receptors.     

Palaeoecological and phytosociological reconstruction of precultural vegetation in the Bílý Kříž area,the Moravskoslezské Beskydy Mts,Slovak Republic

A combined palynological and phytosociological methodological approach was used in the reconstruction of precultural forest vegetation in a small model area of Bílý Kí-Janík in the Moravskoslezské Beskydy mountains (northwest Slovakia). Two pollen diagrams show the predominance of Fagus sylvatica, Abies alba and Picea abies in past natural mixed forests. The present-day occurrence and distribution of forest herbs allow the differentiation of five past vegetation types with above mentioned forest climax trees covering the area before the beginning of human interference. A map of reconstructed natural vegetation is presented which can be used for revitalization of present secondary spruce plantations, heavily affected by air pollution.     

Determination of the labyrinth in different amphibian species and its correlation with determination of the other ectoderm derivatives

The process of labyrinth determination has been studied in three urodelean and seven anuran species by means of homoplastic transplantation of ear region epidermis, defined as the piece of epidermal layer containing the material of the prospective ear vesicle (labyrinth rudiment). The ear region epidermis was grafted onto the abdominal wall of embryos of the same developmental stage. The earliest stage of operation resulting in ectopic ear vesicle formation was determined, suggesting the appearance of organ-specific properties in the ear ectoderm. These properties were enhanced in the further course of development, as indicated by the frequency of ear vesicle formation and by the volume and degree of complexity that the vesicles reached. The data obtained allowed us to arrange the species studied in a sequence, ranging from most Ranidae and Bufo viridis, in which organ-specific properties appear earlier and are most strongly expressed, to Triturus vulgaris in which their expression is least pronounced. Comparison of properties of the material giving rise to the ear vesicle or to several other ectodermal derivatives led to the conclusion that species-specific differences in determination of different ectodermal rudiments are due to species specific properties of the whole ectoderm. These differences appear to be determined by an evolutionary shift of the beginning of gastrulation towards later cleavage cycles.Deceased in November 1993     

Developmental control of the heat-shock stress regulon in Streptomyces coelicolor

In the differentiating eubacterium Streptomyces coelicolor , nutritional imbalances activate a developmental programme which involves the heat-shock stress regulon. In liquid batch cultures, the growth curve could be separated into four components: rapid growth 1 (RG1), transition (T), rapid growth 2 (RG2) and stationary (S). Patterns of gene expression in cultures subjected to heat shock in various phases were recorded on two-dimensional gels and analysed using advanced statistical methods. The responses of all heat-shock proteins (HSPs) were highly dependent upon the growth phase, thus demonstrating that the four phases of growth were physiologically distinct. For many HSPs, the levels of thermal induction attained were closely related to growth stage-determined levels of synthesis before heat shock, thus supporting the idea that developmental and thermal induction of this stress regulon have common control elements. Cluster analysis identified five groups of HSPs displaying similar kinetics of heat and developmentally induced synthesis, probably reflecting the influence of major regulatory systems. Methods introduced here to analyse the response of groups of genes to multiple simultaneous stimuli should find broad applications to studies of other prokaryotic and eukaryotic regulons.     

Viscosity decrease of pectin and fruit juices catalyzed by pectin lyase from Penicillium italicum in batch and continuous-flow membrane reactors

Summary The extracellular pectin lyase (PNL, E.C. 4.2.2.10) from Penicillium italicum was utilized in batch and confined in a continuous-flow ultrafiltration membrane reactor. The enzyme catalyzed the decrease in viscosity of pectin solutions at pH 6.0 as well as of different fruit juices at their respective pH. PNL decreased the viscosity of pectin solutions in the membrane (60% after 30 min) more than in the batch (46% after 70 min) reactors, but similar viscosity reduction of fruit juices was achieved in both reactors. The enzyme decreased the viscosity of grape, peach and melon juices to different extents, but failed to do so with apple or pear juices. It can be concluded that the utilization of PNL in a membrane reactor appears of interest for the clarification of fruit juices.