Journal
SCIENCE OF THE TOTAL ENVIRONMENT
Volume 729, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.scitotenv.2020.139075
Keywords
GST; Gene Kj-gst; Bioremediation; Gene knockout; Point mutation
Categories
Funding
- National Natural Science Foundation of China [31672051]
- National Key Research and Development Programof China [2016YFD0200203]
- Jilin Scientific and Technological Development Program, China [20190301063NY]
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Microbial degradation is one of the most efficient and reliable ways to remove the residues of Chlorimuron-ethyl in the environments such as soil and water. In this study, a glutathione-s-transferase (GST) gene Kj-gst was cloned from the Chlorimuron-ethyl degrading bacterial strain Klebsiella jilinsis 2N3. Results showed that Kj-gst played a key role in the degradation of Chlorimuron-ethyl by strain 2N3. The mutant with gene Kj-gst knocked out showed reduced relative activity up to 70% comparedwith thewild type in 8 h in culture. After the knockout gene was complemented, the degradation ability of the complement mutant was essentially comparable to that of thewild type. The protein Kj-GST (50 mu g) obtained from the gene Kj-gst expressed and purified in E. coli strain BL21(DE3) was capable of degrading Chlorimuron-ethyl with an initial concentration of 50 mg/mL by 42.91% under the optimal conditions (15 degrees C and pH= 7). Point mutation experiments on a glycine located at position 101 (Glu101) confirmed that the H site of glutathione (GSH) is the key component in Kj-GST for degrading Chlorimuron-ethyl. We conclude that Kj-GST is demonstrated for the first time to degrade Chlorimuron-ethyl with its main functional site identified at the H site of GSH, shedding insight to revealing the molecular mechanisms of degrading Chlorimuron-ethyl by Klebsiella jilinsis 2N3. (C) 2020 Elsevier B.V. All rights reserved.
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