Pseudomonas lemoignei has five poly(hydroxyalkanoic acid) (PHA) depolymerase genes: A comparative study of bacterial and eukaryotic PHA depolymerases

Four polyhydroxyalkanoate (PHA) depolymerases were purified from the culture fluid ofPseudomonas lemoignei: poly(3-hydroxybutyrate) (PHB), depolymerase A (M _r , 55,000), and PHB depolymerase B (M _r , 67,000) were specific for PHB and copolymers of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) as substrates. The third depolymerase additionally hydrolyzed poly(3-hydroxyvalerate) (PHV) at high rates (PHV depolymerase;M _r , 54,000). The N-terminal amino acid sequences of the three purified proteins, of a fourth partially purified depolymerase (PHB depolymerase C), and of the PHB depolymerases ofComamonas sp. were determined. Four PHA depolymerase genes ofP. lemoignei (phaZ1,phaZ2,phaZ3, andphaZ4) have been cloned inEscherichia coli, and the nucleotide sequence ofphaZ1 has been determined recently (D. Jendrossek, B. Müller, and H. G. Schlegel,Eur. J. Biochem. 218, 701–710, 1993). In this study the nucleotide sequences ofphaZ2 andphaZ3 were determined.PhaZ1,phaZ2, andphaZ4 were identified to encode PHB depolymerase C, PHB depolymerase B, and PHV depolymerase, respectively.PhaZ3 coded for a novel PHB depolymerase ofP. lemoignei, named PHB depolymerase D. None of the four genes harbored the PHB depolymerase A gene, which is predicted to be encoded by a fifth depolymerase gene ofP. lemoignei (phaZ5) and which has not been cloned yet. The deduced amino acid sequences ofphaZ1–phaZ3 revealed high homologies to each other (68–72%) and medium homologies to the PHB depolymerase gene ofAlcaligenes faecalis T₁ (25–34%). Typical leader peptide amino acid sequences, lipase consensus sequences (Gly-Xaa-Ser-Xaa-Gly), and unusually high proportions of threonine near the C terminus were found in PhaZ1, PhaZ2, and PhaZ3. Considering the biochemical data of the purified proteins and the amino acid sequences, PHA depolymerases ofP. lemoignei are most probably serine hydrolases containing a catalytical triad of Asp, His, and Ser similar to that of lipases. A comparison of biochemical and genetic data of various eubacterial and one eukaryotic PHA depolymerases is provided also.Paper presented at the Bio/Environmentally Degradable Polymer Society—Second National Meeting, August 19–21, 1993, Chicago, Illinois.     

Fixpunkte stetiger Abbildungen Mathematische Theorie und Anwendungen

The purpose of this paper is to give an idea of modern mathematics and of its contribution to science as a whole. As an example to illustrate this the theory of fixed points of continuous mappings has been chosen, because it allows to explain recent mathematical achievements to non-mathematicians without lengthy preparations and because it has several applications. We shall sketch such applications in biology, numerical computation, economics and hydrodynamics.     

Separation of chlorinated hydrocarbons and organophosphorus, pyrethroid pesticides by silicagel fractionation chromatography and their simultaneous determination by GC-MS

A silicagel fractionation procedure for environmental sample extracts, which separates chlorinated hydrocarbons(CHCs) and organophosphorus, pyrethroid pesticides into two groups for subsequent instrumental analysis, was developed in this study. This method was achieved by optimizing the fraction cut-off volume of elution and different solvents. Using fully activated silica gel and cut-off CHCs collection after 10ml 10% dichloromethane (DCM) in n-hexane passing through the column resulted in satisfactory separation of CHCs and organophosphorus, pyrethroid pesticides. This procedure had a higher reliability for CHCs than for organophosphorus, pyrethroid pesticides, because there is a relatively reliable recovery for CHCs. This approach is less expensive due to reducing sample pre-treatment time and solvent consumption.     

DEHP (Di(2-ethylhexyl)phthalat)

Background, Goal and Scope

The plasticiser DEHP is suspected to affect the human reproductive system as an endocrine disruptor (Latini et al. 2004). A meta-analysis was performed to compare annual production amounts and daily uptake of DEHP.

Methods

For this purpose, production data were used from annual publications of the Federal Statistical German Agency (Statistiches Bundesamt) and daily uptake values on the basis of urinary secretion of DEHP metabolites from Wittassek et al. (2007a).

Results

It is shown that the daily uptake of DEHP by German university students is strictly and almost perfectly correlated with the industrial production of DEHP in Germany (correlation coefficient >0.9).

Discussion

The annual production amounts are reflected by the daily uptake values. When the production amount is high, the uptake increases; when the production amount is low, the uptake decreases.

Conclusions

This strict correlation allows the extrapolation of daily DEHP uptake for the 1970s years. On this basis it can be assumed that, at least for a part of the German population, the TDI of 50 μg/kg BW/day was exceeded. Because of their higher susceptibility and, therefore, higher uptake rate (Becker et al. 2004), a considerable exposure for infants might have occurred in these years.     

Responses of a bacterial pathogen to phosphorus limitation of its aquatic invertebrate host

Host nutrition is thought to affect the establishment, persistence, and severity of pathogenic infections. Nutrient-deficient foods possibly benefit pathogens by constraining host immune function or benefit hosts by limiting parasite growth and reproduction. However, the effects of poor elemental food quality on a host's susceptibility to infection and disease have received little study. Here we show that the bacterial microparasite Pasteuria ramosa is affected by the elemental nutrition of its aquatic invertebrate host, Daphnia magna. We found that high food carbon:phosphorus (C:P) ratios significantly reduced infection rates of Pasteuria in Daphnia and led to lower within-host pathogen multiplication. In addition, greater virulent effects of bacterial infection on host reproduction were found in Daphnia-consuming P-deficient food. Poor Daphnia elemental nutrition thus reduced the growth and reproduction of its bacterial parasite, Pasteuria. The effects of poor host nutrition on the pathogen were further evidenced by Pasteuria's greater inhibition of reproduction in P-limited Daphnia. Our results provide strong evidence that elemental food quality can significantly influence the incidence and intensity of infectious disease in invertebrate hosts.     

Online first publication of <Emphasis Type="Italic">Journal of material cycles and waste management</Emphasis>

Announcement

Online first publication of Journal of material cycles and waste management     

Ethylenediurea (EDU): a research tool for assessment and verification of the effects of ground level ozone on plants under natural conditions

Ethylenediurea (EDU) has been widely used to prevent ozone (O₃) injury and crop losses in crop plants and growth reductions in forest trees. Successful use requires establishing a dose/response curve for EDU and the proposed plant in the absence of O₃ and in the presence of O₃ before initiating multiple applications to prevent O₃ injury. EDU can be used to verify foliar O₃ symptoms in the field, and to screen plants for sensitivity to O₃ under ambient conditions. Despite considerable research, the mode of action of EDU remains elusive. Additional research on the mode of action of EDU in suppressing O₃ injury in plants may also be helpful in understanding the mode of action of O₃ in causing injury in plants.     

Degradative fate of 3-chlorocarbazole and 3,6-dichlorocarbazole in soil

Background, aim, and scope  

3-Chlorocarbazole and 3,6-dichlorocarbazole were isolated from Bavarian soils. The stereospecific formation of the isomers of these chlorinated carbazols can be explained by quantum mechanical calculations using the DFT method. It was shown that chlorination of carbazole and 3-chlorocarbazole respectively is preferred via the sigma-complexes 3-chlorocarbazole and 3,6-dichlorocarbazole as the most stable products. The dioxin-like toxicological potential of 3,6-dichlorocarbazole, determined by the Micro-EROD Test, is in the range of some picogram TCDD equivalents per milligram carbazole. The degradative fate of 3-chlorocarbazole and 3,6-dichlorocarbazole was analysed within a long-term study (57 days) in soil.