Henryk Kołoczek

Effects of nickel on the chloroplasts of the duckweeds Spirodela polyrhiza and Lemna minor and their possible use in biomonitoring and phytoremediation.

Toxic effects of Ni(2+) on the chloroplasts of the two duckweed species Spirodela polyrhiza, clone SJ and Lemna minor, clone St were investigated according to the ISO 20079 protocol. Ni(2+) induced a transition from chloroplasts to chloro-amyloplasts and amylo-chloroplasts, but not to gerontoplasts, as shown by electron microscopy. The contents of the chlorophylls a and b decreased strongly, whereas that of carotenoids remained approximately constant. Most striking was, however, the accumulation of transitory starch. Bell-shaped dose-response curves showed that Spirodela and Lemna amassed maximum starch contents of approximately 10% and 7%, respectively, on a fresh weight basis. Because Ni(2+) in the concentrations applied does not stimulate photosynthesis, the Ni(2+)-induced starch accumulation indicates that the export of carbohydrates out of the plastids decreased, most probably due to the lower demand of the rest of the cells as a result of the Ni(2+)-dependent inhibition of growth. The half-maximal concentrations for inhibition of the fresh weight increase over the 7-day test period were 3.7 microM and 6.6 microM for Spirodela and Lemna, respectively: Spirodela was thus somewhat more sensitive to the heavy metal. Both species accumulated approximately 3g of Ni(2+) per kg of dry weight after application of 100 microM NiCl(2). Because of their high sensitivity to phytotoxic effects, however, Spirodela and Lemna do not appear to be particularly suitable for phytoremediation of Ni(2+)-contaminated waste water. The high sensitivity to Ni(2+) makes them instead a suitable system for ecotoxicological testing in accordance with the ISO 20079 protocol.

Chromium(VI) bioremediation by aquatic macrophyte Callitriche cophocarpa Sendtn.

Callitriche cophocarpa (water-starwort)--aquatic widespread macrophyte--was found to be an excellent chromium accumulator. The plants were exposed to various chromium(VI) concentration ranging from 50 to 700 microM in a hydroponic culture up to ca. 3 weeks. Physiological conditions of shoots were monitored via measuring potential photosynthesis quantum efficiency (F(v)/F(m)) and photosynthetic pigment contents. Additionally, the structure of leaves was analyzed using optical and atomic force microscopy (AFM). It has been shown that plants grown in 50 microM Cr(VI) solution exhibited photosynthetic activity and shoot and leaf morphology similar to control plants. Moreover, at the same time the average Cr concentration in their shoots reached about 470 mg kg(-1)d.w. after 10d and up to 1000 mg kg(-1)d.w. after 3 weeks of culture while in control plants did not exceed a few mgkg(-1)d.w. Our results point to Callitriche cophocarpa as a very promising species to be used in the investigation of chromium(VI) phytoremediation mechanisms as well as a good candidate for wastewaters remediation purpose.

Formaldehyde and methanol biodegradation with the methylotrophic yeast Hansenula polymorpha. An application to real wastewater treatment

The application of methylotrophic yeast Hansenula polymorpha to the treatment of methanol and formaldehyde-containing wastewater was experimentally verified. Avariety of real wastewater samples originating from chemical industry effluent were examined. The yeast cell culture could grow in the wastewater environment, revealing low trophic requirements and a very high adaptation potential to poor cultivation conditions.The proliferation of cells was accompanied by a concomitant xenobiotic biodegradation. Grown, preadapted cellular suspension at a density of about 1 × 107 cells/ml proved to be able to utilize formaldehyde present in wastewater at concentrations up to1750 mg/l, levels toxic to most microorganisms. The biological waste treatment method presented shows the enhanced potential by means of specific enzymatic activities of monocarbonic compound oxidations through methylotrophic pathway reactions. The need to obtain mutants highly resistant to formaldehyde has also been rationalized.

Formaldehyde and methanol biodegradation with the methylotrophic yeast Hansenula polymorpha in a model wastewater system

In search of the optimal way to reduce the hazards of environmental contamination by formaldehyde (FD) and methanol the use of unconventional yeasts is proposed as exemplified by the methylotrophic yeast Hansenula polymorpha. In a very simplified environment of a model wastewater solution, H. polymorpha cells were able to grow on, and metabolize formaldehyde and methanol, applied as sole carbon sources, at concentrations typical for wastewaters of the chemical industry. Several experimental conditions were tested for cell growth and biodegradation kinetics. It was found that the yeast culture inoculated at low cell density was able to grow on initial FD levels up to 400mg/l and the biomass yield was dependent on both, the amount of total carbon added and the physiological state of the cells. When high density of pre-adapted cell culture was used, the methylotrophs were fully viable and able to degrade formaldehyde present at initial concentrations up to 700 mg/l. The maximum limiting FD consumption rate was determined as approx. 400 mg/1 per hour. Methanol, at concentrations up to 2%, was easily utilized and did not have a negative effect on cell growth and respiration. It is suggested that in real wastewaters the eukaryotic microorganisms--in contrast to bacteria--might reveal greater adaptation potential to toxic levels of formaldehyde as well as to other wastewater constituents.

Biodegradation of formaldehyde and its derivatives in industrial wastewater with methylotrophic yeast Hansenula polymorpha and with the yeast-bioaugmented activated sludge.

Methylotrophic yeast Hansenula polymorphawere shown to cooperate with activated sludgefrom biological wastewater treatment stations,enhancing substantially its potential tobiodegrade formaldehyde in industrial wastewater. After integration with yeast cells the modified sludge retained its original structure and activity whereas its resistance to elevated formaldehyde concentrations was significantly improved. The applicability of the yeast in the utilization of formaldehyde derivatives, as exemplified by urotropine and trioxane, was also investigated. The treatment of urotropine-containing wastewater with methylotrophic yeast was found to be effective at acidic conditions (pH below 5.5). Trioxane was not degraded due to the stability of an ether bond which made the molecule recalcitrant to oxidation via methylotrophic pathway reactions. It is concluded that the yeast species may be applied to treat wastewater containing formaldehyde and some of its derivatives as either monocultures or as an integrated, specialized element of the activated sludge biocenosis.

Fluorescence-detected polymerization kinetics of human alpha 1-antitrypsin.

The time dependence of the humanα1-antitrypsin polymerization process was studied by means of the intrinsic fluorescence stopped-flow technique as well as the fluorescence-quenching-resolved spectra (FQRS) method and native PAGE. The polymerization was induced by mild denaturing conditions (1 M GuHCl) and temperature. The data show that the dimer formation reaction under mild conditions was followed by an increase of fluorescence intensity. This phenomenon is highly temperature sensitive. The structure ofα1-antitrypsin dimer resembles the conformation of antithrombin III dimer. In the presence of the denaturant the polymerization process is mainly limited to the dimer state. Theα1-antitrypsin activity measurements confirm monomer-to-dimer transition under these conditions. These results are in contrast to the polymerization process induced by temperature, where the dimer state is an intermediate step leading to long-chain polymers. On the basis of stopped-flow and electrophoretic data it is suggested that both C-sheet as well as A-sheet mechanisms contribute to the polymerization process under mild conditions.