חיפוש מתקדם
תחביר
חפש...
הספר "אוצר וולקני"
אודות
תנאי שימוש
ניהול
קהילה:
אסיף מאגר המחקר החקלאי
פותח על ידי קלירמאש פתרונות בע"מ -
Atrazine Hydrolysis by a Bacterial Enzyme
Year:
1998
Source of publication :
ACS Symposium Series
Authors :
מנדלבאום, רפי
;
.
Volume :
683
Co-Authors:
Wackett, L.P., Department of Biochemistry, Bioprocess Technology Institute, University of Minnesota, St. Paul, MN 55108, United States, Ctr. Biodegradation Res. and Info., University of Minnesota, St. Paul, MN 55108, United States
Sadowsky, M.J., Ctr. Biodegradation Res. and Info., University of Minnesota, St. Paul, MN 55108, United States, Dept. of Soil, Water and Climate, University of Minnesota, St. Paul, MN 55108, United States
De Souza, M., Department of Biochemistry, Bioprocess Technology Institute, University of Minnesota, St. Paul, MN 55108, United States, Ctr. Biodegradation Res. and Info., University of Minnesota, St. Paul, MN 55108, United States
Mandelbaum, R.T., Volcani Research Institute, Bet-Dagan 50250, Israel
Facilitators :
From page:
82
To page:
87
(
Total pages:
6
)
Abstract:
Atrazine, 2-chloro-4-(ethylamino)-6-(isopropylamino)-1,3,5-triazine, is metabolized relatively slowly in natural soils and waters by resident microorganisms. Recently, several atrazine-degrading bacterial pure cultures were isolated and the molecular basis of bacterial atrazine metabolism is now beginning to be revealed. Pseudomonas sp. strain ADP was isolated from a herbicide spill site for its ability to use atrazine as the sole source of nitrogen for growth. Atrazine metabolism also liberated the triazine ring carbon atoms as carbon dioxide. Hydroxyatrazine was detected transiently in the growth medium during the course of atrazine metabolism. Previously, hydroxyatrazine was proposed to be derived solely from abiotic hydrolysis catalyzed by soil organic matter and clays. The gene encoding the enzymatic hydrolysis of atrazine by Pseudomonas sp. ADP was cloned and expressed in Escherichia coll. Cell-free atrazine hydrolysis activity in the recombinant E. coli strain was determined by high pressure liquid chromatography. The enzyme, atrazine chlorohydrolase, was purified to homogeneity using ammonium sulfate precipitation and ion exchange chromatography. The purified chlorohydrolase showed a single band on denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis corresponding to a subunit molecular weight of 60,000. Gene sequencing data yielded a molecular weight of 52,421. Gel filtration chromatography indicated a holoenzyme molecular weight of 240,000 consistent with an α4 or α5 subunit stoichometry. In [18O]-H2O, atrazine chlorohydrolase yielded [18O]-hydroxyatrazine quantitatively. In control experiments incubated and analyzed under the same conditions, [18O] from H2O did not exchange into hydroxyatrazine. These data are consistent with enzymatic hydrolysis of atrazine. Other bacteria were also demonstrated to catalyze atrazine hydrolysis, suggesting this biologically-mediated reaction is widespread in soil and water.
Note:
Related Files :
עוד תגיות
תוכן קשור
More details
DOI :
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
24487
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:08
Scientific Publication
Atrazine Hydrolysis by a Bacterial Enzyme
683
Wackett, L.P., Department of Biochemistry, Bioprocess Technology Institute, University of Minnesota, St. Paul, MN 55108, United States, Ctr. Biodegradation Res. and Info., University of Minnesota, St. Paul, MN 55108, United States
Sadowsky, M.J., Ctr. Biodegradation Res. and Info., University of Minnesota, St. Paul, MN 55108, United States, Dept. of Soil, Water and Climate, University of Minnesota, St. Paul, MN 55108, United States
De Souza, M., Department of Biochemistry, Bioprocess Technology Institute, University of Minnesota, St. Paul, MN 55108, United States, Ctr. Biodegradation Res. and Info., University of Minnesota, St. Paul, MN 55108, United States
Mandelbaum, R.T., Volcani Research Institute, Bet-Dagan 50250, Israel
Atrazine Hydrolysis by a Bacterial Enzyme
Atrazine, 2-chloro-4-(ethylamino)-6-(isopropylamino)-1,3,5-triazine, is metabolized relatively slowly in natural soils and waters by resident microorganisms. Recently, several atrazine-degrading bacterial pure cultures were isolated and the molecular basis of bacterial atrazine metabolism is now beginning to be revealed. Pseudomonas sp. strain ADP was isolated from a herbicide spill site for its ability to use atrazine as the sole source of nitrogen for growth. Atrazine metabolism also liberated the triazine ring carbon atoms as carbon dioxide. Hydroxyatrazine was detected transiently in the growth medium during the course of atrazine metabolism. Previously, hydroxyatrazine was proposed to be derived solely from abiotic hydrolysis catalyzed by soil organic matter and clays. The gene encoding the enzymatic hydrolysis of atrazine by Pseudomonas sp. ADP was cloned and expressed in Escherichia coll. Cell-free atrazine hydrolysis activity in the recombinant E. coli strain was determined by high pressure liquid chromatography. The enzyme, atrazine chlorohydrolase, was purified to homogeneity using ammonium sulfate precipitation and ion exchange chromatography. The purified chlorohydrolase showed a single band on denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis corresponding to a subunit molecular weight of 60,000. Gene sequencing data yielded a molecular weight of 52,421. Gel filtration chromatography indicated a holoenzyme molecular weight of 240,000 consistent with an α4 or α5 subunit stoichometry. In [18O]-H2O, atrazine chlorohydrolase yielded [18O]-hydroxyatrazine quantitatively. In control experiments incubated and analyzed under the same conditions, [18O] from H2O did not exchange into hydroxyatrazine. These data are consistent with enzymatic hydrolysis of atrazine. Other bacteria were also demonstrated to catalyze atrazine hydrolysis, suggesting this biologically-mediated reaction is widespread in soil and water.
Scientific Publication
נגישות
menu      
You may also be interested in