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Biotransformation of 4-nitrophenol by Corynebacterium sp. strain 83
Abstracts Bromoxynil (3,5-dibromo-4-hydroxybenzonitrile), ioxynil (3,5diiodo-4-hydroxybenzonitrile) and dichlobenil (2,6-dichlorobenzonitrile) are widely used halogenated arylnitrile herbicides. The main way of their removal from the environment is their biodegradation by soil microorganisms. A new A. radiobacter 8/4 strain was isolated from bromoxynil contaminated soil in our laboratory by soil enrichment technique. This strain produces nitrile hydratase enzyme. transforming aromatic nitriles to their corresponding amides. A radiobacter 8/4 strain was studied from the point of view of preparation of alginate. carrageenan and pectate immobilized biocatalysts for decontamination of waste water containing aromatic nitriles, e.g. bromoxynil. ioxynil or dichlobenil, or for production of aromatic amides of high purity for pharmacy, etc.
Activities of Ligninolytic Enzymes in White-Rot Fungi Growing in Lignocellulosic Substrate and in Soil P. Erbanova“, e. Novotnj@, F. Zadraii? and V. SaSek” “Lahorutory Experimental Mycology. Institute of’ Microbiology. Czech Academy of Sciences, 142 20 Prague 4, Videriskd 1083. Czech Republic, and hFA L Braunschweig. Bundesallee SO, 38116, Braurtschweig. German? Ligninolytic enzymes of white-rot fungi degrade besides lignin and also other aromatic compounds, including poly- and chloroaromatics. which often represent important soil pollutants. Thus a major prerequisite for successful bioremediation is the presence of these enzymes in the contaminated soil in sufficient quantities. Three species of white-rot fungi, Phanerochaete chr~sosporium, Trametes versicolor and Pleurotus ostreatus, were cultivated on straw in aerated tube reactors filled with soil. The levels of individual ligninolytic enzymes in straw and soil and fungal growth into soil (ergosterol method) were estimated in weekly intervals. The respective fungal growth yields of the above-mentioned fungi were 6.2. 2.2 and I l.Opg ergosterol per g soil. The activities of MnP. LiP and lactase were highest in T. versicolor and lowest in P. chrysosporium. in both straw and soil. The enzyme activities in straw (0.64.1 nkat/g dry matter) exceeded those in soil (O-O.7 nkatlg dry matter).
Biotransformation of 4-Nitrophenol Covynebacterium sp. Strain g/3
by
D. HorikovP and L. KotouCkovP Department of’Microbiolog_v. MU Brno. Tvrdeho 14, 602 00 Brno. CR, Czech Republic Nitrophenols are widely used in the manufacture of pesticides, explosives and dyes. They are classified as common pollutants in natural ecosystem. p-Nitrophenol (PNP) is an important intermediate of microbial or photo-induced degradation of the organophosphorous insecticides. Different kinds of microorganisms are responsible for remove of PNP from contaminated soil and water. We have isolated PNP-degrading strain Corynebacterium sp. S/3 from polluted soil. This organism transforms PNP under aerobic conditions to red product (,I max. 510nm). Bacteria were grown in mineral medium supplemented with pepton and PNP in concentrations about 7-l 50 mg I-‘. PNP concentration was monitored spectrophotometrically by A 398 (pH above 8). Growth of the isolated strain Coryrebucterium sp. 8/3 is suppressed partially if high concentration of PNP in medium is used (75 or 150mg I-‘, respectively). But values of A 508 were found the highest for those high initially concentrations of PNP. Resting-cells experiment has shown temperatureoptimum for degradation 25°C and value of pH 8. This value
249
is optimum for growth of cells too. Cells were activated with preincubation in medium supplemented with 50 mgl-’ PNP. The length of preincubation 72 h was found to be more effective than 24-h preincubation.
Biodegradation of Carbamate Juvenile Hormone Analogue by Soil Microorganisms M. Hronova”, T. Ruml’ and R. Tykva* “Department qf Biochemistry and Microbiology,. Institute of Chemical Technology, Prague, Czech Republic, and hInstitute of Organic Chemistry and Biochemistr_v, Academy of Sciences, Prague, Czech Republic The application of new pesticides can cause serious environmental problems. Therefore potential stability of the recently developed [‘4C]-labelled juvenile hormone analogue W-328 in the water ecosystem was tested. Total of 31 microorganisms was isolated from two different soil samples. All the isolates were tested for their ability to decompose W328. The degradation products were estimated using radioHPLC. One bacteria1 strain. one yeast and four mold isolates exhibited the degradation activity. All of these strains were characterised and their growth characteristics such as pH and temperature dependencies were determined. The results obtained will be discussed with respect to soil differences.
Anaerobic PCB Biodegradation in Contaminated Fats - Evaluation of Laboratory Experiments J. Burkbard, P. Sliacka, J. ZibranskP and M. Dobhyos Fact&v of Environmental Technolog+v, ICT Prague, Techmcka 5, 166 28 Prague, Czech Republic The results of anaerobic PCB degradation in contaminated fats are presented. The process was carried out in batch laboratory anaerobic reactors inoculated with microbial consortia from an anaerobic tanks of a waste water treatment plant. The extent of degradation was followed by capillary gas chromatography using ECD and various external standards. Experimental data are expressed in three courses: - total PCB amount after biodegradation (the sum of the indicator congeners), - leftover of individual congeners, ~ normalized chromatograms of PCB fractions. Comparison of the results obtained by these three interpretation techniques is examined. The experiments proved the ability of employed microbial consortia to degrade PCB under the anaerobic conditions,
The Evaluation of the Course of the Biodegradation of PCB in the Water Suspension of Contaminated Soil J. Burkbard”, B. Polikovi”, K. Demnero& and J. PazlarovP’ UDepartment of Environmental Chemistry, ICT Prague. Technicka 5, 166 28 Prague 6, Czech Republic, and hDepartment of’ Biochemistry and Microbiology. ICT Prague, Technickd 5. 166 28 Prague 6, Czech Republic A method for the evaluation of biodegradation course in laboratory conditions is described. The analytical procedure consists of extraction of the whole content of the cultivation flask and gas chromatographic analysis of the extract on capillary column with the EC-detector and various external standards. Presented comparison of the three expressions of experimental data the residue after biodegradation of individual congeners. of total residue of PCB fraction (as a sum of indicator congeners) and normalized chromatograms of PCB fractions ~ gives a large scale of informations about the course and the results of the biodegradation.
Activities of Ligninolytic Enzymes in White-Rot Fungi Growing in Lignocellulosic Substrate and in Soil P. Erbanova“, e. Novotnj@, F. Zadraii? and V. SaSek” “Lahorutory Experimental Mycology. Institute of’ Microbiology. Czech Academy of Sciences, 142 20 Prague 4, Videriskd 1083. Czech Republic, and hFA L Braunschweig. Bundesallee SO, 38116, Braurtschweig. German? Ligninolytic enzymes of white-rot fungi degrade besides lignin and also other aromatic compounds, including poly- and chloroaromatics. which often represent important soil pollutants. Thus a major prerequisite for successful bioremediation is the presence of these enzymes in the contaminated soil in sufficient quantities. Three species of white-rot fungi, Phanerochaete chr~sosporium, Trametes versicolor and Pleurotus ostreatus, were cultivated on straw in aerated tube reactors filled with soil. The levels of individual ligninolytic enzymes in straw and soil and fungal growth into soil (ergosterol method) were estimated in weekly intervals. The respective fungal growth yields of the above-mentioned fungi were 6.2. 2.2 and I l.Opg ergosterol per g soil. The activities of MnP. LiP and lactase were highest in T. versicolor and lowest in P. chrysosporium. in both straw and soil. The enzyme activities in straw (0.64.1 nkat/g dry matter) exceeded those in soil (O-O.7 nkatlg dry matter).
Biotransformation of 4-Nitrophenol Covynebacterium sp. Strain g/3
by
D. HorikovP and L. KotouCkovP Department of’Microbiolog_v. MU Brno. Tvrdeho 14, 602 00 Brno. CR, Czech Republic Nitrophenols are widely used in the manufacture of pesticides, explosives and dyes. They are classified as common pollutants in natural ecosystem. p-Nitrophenol (PNP) is an important intermediate of microbial or photo-induced degradation of the organophosphorous insecticides. Different kinds of microorganisms are responsible for remove of PNP from contaminated soil and water. We have isolated PNP-degrading strain Corynebacterium sp. S/3 from polluted soil. This organism transforms PNP under aerobic conditions to red product (,I max. 510nm). Bacteria were grown in mineral medium supplemented with pepton and PNP in concentrations about 7-l 50 mg I-‘. PNP concentration was monitored spectrophotometrically by A 398 (pH above 8). Growth of the isolated strain Coryrebucterium sp. 8/3 is suppressed partially if high concentration of PNP in medium is used (75 or 150mg I-‘, respectively). But values of A 508 were found the highest for those high initially concentrations of PNP. Resting-cells experiment has shown temperatureoptimum for degradation 25°C and value of pH 8. This value
249
is optimum for growth of cells too. Cells were activated with preincubation in medium supplemented with 50 mgl-’ PNP. The length of preincubation 72 h was found to be more effective than 24-h preincubation.
Biodegradation of Carbamate Juvenile Hormone Analogue by Soil Microorganisms M. Hronova”, T. Ruml’ and R. Tykva* “Department qf Biochemistry and Microbiology,. Institute of Chemical Technology, Prague, Czech Republic, and hInstitute of Organic Chemistry and Biochemistr_v, Academy of Sciences, Prague, Czech Republic The application of new pesticides can cause serious environmental problems. Therefore potential stability of the recently developed [‘4C]-labelled juvenile hormone analogue W-328 in the water ecosystem was tested. Total of 31 microorganisms was isolated from two different soil samples. All the isolates were tested for their ability to decompose W328. The degradation products were estimated using radioHPLC. One bacteria1 strain. one yeast and four mold isolates exhibited the degradation activity. All of these strains were characterised and their growth characteristics such as pH and temperature dependencies were determined. The results obtained will be discussed with respect to soil differences.
Anaerobic PCB Biodegradation in Contaminated Fats - Evaluation of Laboratory Experiments J. Burkbard, P. Sliacka, J. ZibranskP and M. Dobhyos Fact&v of Environmental Technolog+v, ICT Prague, Techmcka 5, 166 28 Prague, Czech Republic The results of anaerobic PCB degradation in contaminated fats are presented. The process was carried out in batch laboratory anaerobic reactors inoculated with microbial consortia from an anaerobic tanks of a waste water treatment plant. The extent of degradation was followed by capillary gas chromatography using ECD and various external standards. Experimental data are expressed in three courses: - total PCB amount after biodegradation (the sum of the indicator congeners), - leftover of individual congeners, ~ normalized chromatograms of PCB fractions. Comparison of the results obtained by these three interpretation techniques is examined. The experiments proved the ability of employed microbial consortia to degrade PCB under the anaerobic conditions,
The Evaluation of the Course of the Biodegradation of PCB in the Water Suspension of Contaminated Soil J. Burkbard”, B. Polikovi”, K. Demnero& and J. PazlarovP’ UDepartment of Environmental Chemistry, ICT Prague. Technicka 5, 166 28 Prague 6, Czech Republic, and hDepartment of’ Biochemistry and Microbiology. ICT Prague, Technickd 5. 166 28 Prague 6, Czech Republic A method for the evaluation of biodegradation course in laboratory conditions is described. The analytical procedure consists of extraction of the whole content of the cultivation flask and gas chromatographic analysis of the extract on capillary column with the EC-detector and various external standards. Presented comparison of the three expressions of experimental data the residue after biodegradation of individual congeners. of total residue of PCB fraction (as a sum of indicator congeners) and normalized chromatograms of PCB fractions ~ gives a large scale of informations about the course and the results of the biodegradation.