Lipase-catalyzed synthesis of lysophosphatidic acid in a solvent free system

Summary The direct, lipase-catalyzed esterifications of glycerol-3-phosphate in an organic solvent system and in a solvent free system were carried out. In a solvent free system only, LPA synthesis could be achieved within the acceptable reaction time. Open reaction system was preferable to closed reaction system for LPA synthesis. Yield of LPA isolated by silica gel column chromatography was 32.3%.     

Temperature coefficients of peptides dissolved in hexafluoroisopropanol monitor distortions of helices

Temperature coefficients are widely used as an indication of solvent accessibility to amide protons. Low temperature coefficients are related to low accessibility and are often interpreted as evidence for intramolecular hydrogen bonding. Conformational shifts, i.e. the difference between chemical shifts of a particular residue in a structured and in a random-coil conformation, provide information on secondary structure. In particular, negative CH conformational shifts are often used to delineate the extent of helical stretches. NH conformational shifts show large oscillations within a helix that have been interpreted as the result of helix distortions affecting hydrogen bond lengths. In the course of the study of different peptides that adopt a helical structure in the presence of the structure-inducing solvent hexafluoroisopropanol (HFIP), we have found a strong correlation between temperature coefficients and amide conformational shifts. However, contrary to the initial expectations, lower temperature coefficients were associated to amide protons involved in longer, and presumably weaker, hydrogen bonds. The correlation can be explained, however, assuming that, in helical peptides dissolved in HFIP, temperature affects the chemical shift of amide protons mainly by changing the average length of intramolecular hydrogen bonds and changes in solvent accessibility play only a secondary role under these experimental conditions. The pattern of temperature coefficients in helical peptides can therefore be used to identify short or long hydrogen bonds causing bending of the helix axis.     

Regulation of inositol monophosphatase in Saccharomyces cerevisiae

Inositol monophosphatase is a key enzyme in the de novo biosynthesis of inositol and in the phosphoinositide second-messenger signalling pathway. Inhibition of this enzyme is a proposed mechanism for lithium's pharmacological action in bipolar illness (manic depression). Very little is known about how expression of this enzyme is regulated. Because the yeast Saccharomyces cerevisiae has been shown to be an excellent model system in which to understand the regulation of inositol metabolism, we characterized inositol monophosphatase in this yeast. Lithium inhibited monophosphatase activity in vitro . Growth in the presence of inositol resulted in increased expression of the enzyme in vivo , although inositol had no effect on enzyme activity in vitro . The inositol effect was apparent when cells were grown in glucose but not in glycerol/ethanol. Monophosphatase activity was derepressed as cells entered stationary phase. This effect was apparent only during growth in glucose plus inositol. The results demonstrate that S. cerevisiae monophosphatase is inhibited by lithium and regulated by factors affecting phospholipid biosynthesis.     

Grenzen der Kompensation: Energiebudgets von Ringelg?nsen (Branta b. bernicla) — die Wirkung von St?rreizen

Zusammenfassung Der Einfluß von Störreizen auf das Energiebudget von Ringelgänsen wurde im Frühjahr 1991 in zwei Vorland-Salzwiesen des Nationalparks Schleswig-Holsteinisches Wattenmeer vergleichend untersucht. Die Häufigkeit störreizbedingter Reaktionen der Gänse (Störreizhäufigkeit) wurde als Index für die anthropogene Beeinflussung der Gebiete herangezogen. Die Beeinflussung war in Westerhever mit 1,5±0,7 Reaktionen/h signifikant größer als vor dem Norderheverkoog (1,0±0,6 Reaktionen/h). Mit zunehmender Aktivitätszeit wurde in beiden Gebieten im Verlauf des Frühjahres mehr umsetzbare Energie aufgenommen. In Westerhever haben die Gänse im Vergleich zu denen vor dem Norderheverkoog mit Ausnahme des Mai jeweils mehr umsetzbare Energie aufgenommen. Die energetischen Kosten (DEE) der Gänse wurden anhand der Zeit-Energie-Budget Methode und über die gemessene Körpermasseentwicklung von Fänglingen berechnet. Beide Methoden erzielten vergleichbare Ergebnisse; die Unterschiede betrugen je nach Gebiet 0,4 bis 4,5 %. In Westerhever verzeichneten die Gänse im Vergleich zu denen vor dem Norderheverkoog im Monatsmittel höhere energetische Kosten. Aus dem budgetierten Energieüberschuß (E) wurde die theoretisch mögliche Reservestoffanlagerung (Änderung der Körpermasse) der Gänse berechnet. In Westerhever konnten die Gänse im Verlauf des Frühjahres 357 g Körperreserven anlagern. Bei den Gänsen vor dem Norderheverkoog waren es 399 g. Der Unterschied betrug 11,7 %, war aber nicht signifikant. Anhand von Fangdaten ist bekannt, daß Ringelgänse im langjährigen Mittel Körperreserven von 380 bis 400 Gramm anlagern. Die Störreizhäufigkeit des Tages bewirkte eine signifikante Veränderung der Budgetparameter DME und DEE und E auf stündlicher Basis. In Westerhever stieg die Aufnahme an umsetzbarer Energie signifikant mit steigender Störreizhäufigkeit an. Vor dem Norderheverkoog verringerte sich diese jedoch signifikant. Die Energieaufwendungen stiegen in beiden Gebieten signifikant mit steigender Reizhäufigkeit an. Der Überschuß in der Energiebilanz der Gänse in Westerhever war an Tagen mit einer großen Störreizhäufigkeit um 8,7 % gegenüber den Tagen mit einer geringen Reizhäufigkeit verringert. Aufgrund fehlender kompensatorischer Nahrungsaufnahme war der Überschuß bei den Gänsen vor dem Norderheverkoog um maximal 27,5 % vermindert. Die Gänse in Westerhever haben störreizbedingte Zeitverluste bei der Nahrungsaufnahme und erhöhte energetische Kosten durch Verhaltensänderung und eine erhöhte Nahrungsaufnahme pro Zeit kompensiert. Aufgrund physiologischer Zwänge sind sie jedoch in dem stark vom Menschen beeinflußten Gebiet an die Grenzen ihrer Kompensationsmöglichkeit angelangt. Die errechnete Reservestoffanlagerung und auch der Jungvogelanteil im nachfolgenden Herbst waren geringer als bei den Gänsen vor dem Norderheverkoog.

---

Compensatory limits: energy budgets of Brent Geese,Branta b. bernicla, the influence of human disturbance

The impact of human disturbances on the energy budget of Brent Geese,Branta b. bernicla, during spring migration was investigated in two different salt marsh areas within the national park Schleswig-Holsteinisches Wattenmeer, Germany. We used the frequency of disturbance-related reactions of the birds as an indicator of the anthropogenic influencing of the sites. The disturbance frequency was significantly higher in the Westerhever salt marsh (1,5±0,7 reactions/h) than in the Norderheverkoog area (1,0±0,6 reactions/h). Monthly activity and energy budgets of the birds were calculated from March to May, based on several simultaneous day-round field observations per month. The amount of daily metabolizable energy (DME) was calculated via dropping rate and digestability of the food. The daily energy expenditure (DEE) was calculated by means of the time-energy-budget methods and by calculations, using the bodymass changes of caught birds during spring. With both methods very similar results were obtained; they varied between 0,4 to 4,5 % per areas and month. The daily energy budget (Æ E) was calculated from DEE — DME. Due to an increasing daily activity time from March to Mai the DME increased in both areas. Compared to the Norderheverkoog area the DME of the birds in Westerhever was higher in March and April, and similar in Mai. DEE increased in both areas too and was in all months higher in Westerhever than in Norderheverkoog. The calculated bodymass changes of the birds during spring was 357 g for the birds in Westerhever and 399 g for the birds in Norderheverkoog. Bodymass measures from caught birds revealed a mean mass gain of 380 to 400 g during spring. The daily disturbance frequency caused a significant change in the budget parameters DME, DDE and E on a hourly base. In Westerhever DME/h increased, while in Norderheverkoog DME/h decreased significantly with increasing disturbance frequency. At the same time DEE/h increased in both areas significantly. As a consequence E/h was reduced at days with a high disturbance frequency; in Westerhever by 8,7 % and in Norderheverkoog by 27,5%. In the disturbed area the birds compensated the time-loss during feeding and the higher energetic costs during disturbance-related flights by a change in activity pattern and by an increased food consumption per time unit. Based on physiological constraints the birds reached their compensatory abilities. The calculated bodymass gain and the proportion of young birds in the flocks was reduced in autumn.

     

Cloning and characterization of a gene from Pasteurella haemolytica A1 involved in lipopolysaccharide biosynthesis

Abstract A Pasteurella haemolytica A1 gene involved in the biosynthesis of a moiety on the core of the lipopolysaccharide molecule has been cloned and characterized. Escherichia coli clones which carry this gene showed an alteration of its lipopolysaccharide migration profile on tricine SDS-PAGE and exhibited resistance to the core-specific phage U3. In addition, lipopolysaccharide extracted from the E. coli clones was recognized by an anti-corespecific antiserum, but not by antiserum specific for the O antigen of P. haemolytica A1 lipopolysaccharide. Nucleotide sequence analysis of the cloned DNA identified an open reading frame ( lpsA ) coding for a protein of 263 amino acids which showed significant homology with a Haemophilus influenzae type b lipooligosaccharide biosynthesis gene. PCR amplification of genomic DNA, using primers based on the P. haemolytica A1 lpsA sequence, yielded products from only the A biotypes of P. haemolytica .     

Analysis of viability and cell types of macroalgal protoplasts using flow cytometry

The ability to rapidly distinguish viable sub-populations of cells within populations of macroalgal protoplast isolations was demonstrated using flow cytometry. Viable protoplasts from Ulva sp. and Porphyra perforata J. Ag. were distinguished from non-viable protoplasts based on differential fluorescein accumulation. The identities of cortical and epidermal protoplasts from Macrocystis pyrifera (L.) C. Ag. were inferred based on light-scattering and chlorophyll a autofluorescence. Three cell types could be distinguished among protoplasts released from thalli of P. perforata based on chlorophyll a and phycoerythrin autofluorescence. Mixed protoplast populations of Ulva sp. and P. perforata were also discernable based on relative chlorophyll a and phycoerythrin autofluorescence. The ability to screen heterogenous protoplast populations rapidly, combined with the cell sorting capabilities of many flow cytometers, should prove valuable for seaweed biotechnology.     

The maintenance ofBdellovibrio at low prey density

A mathematical model for the interaction ofBdellovibrio and its prey predicted that a relatively high prey density (7×10⁵ cells ml^–1) would be required for the establishment of an equilibrium in a mixed population [8]. The present report shows thatBdellovibrio can be maintained in a continuous culture when the prey cell density is much lower (2–5×10⁴ cells ml^–1), and closer to that of naturally occurring bacterial populations in sea waters.