Effect of culture conditions on the whole cell activity of recombinant Escherichia coli expressing periplasmic organophosphorus hydrolase and cytosolic GroEL/ES chaperone
Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigat...
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| Published in | Biotechnology and bioprocess engineering Vol. 21; no. 4; pp. 502 - 507 |
|---|---|
| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Seoul
The Korean Society for Biotechnology and Bioengineering
01.08.2016
Springer Nature B.V 한국생물공학회 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1226-8372 1976-3816 |
| DOI | 10.1007/s12257-016-0342-y |
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| Abstract | Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant
Escherichia coli
coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES. When cultured at 20°C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 µM CoCl
2
, the recombinant
E. coli
exhibited a specific whole cell OPH activity (U/OD
600
) of ~0.55, which is 2.6-fold higher than that of recombinant
E. coli
cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response. Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates. |
|---|---|
| AbstractList | Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors.
Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES.
When cultured at 20°C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 μM CoCl2, the recombinant E. coli exhibited a specific whole cell OPH activity (U/OD600) of ~0.55, which is 2.6-fold higher than that of recombinant E. coli cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response.
Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates. KCI Citation Count: 2 Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES. When cultured at 20°C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 µM CoCl 2 , the recombinant E. coli exhibited a specific whole cell OPH activity (U/OD 600 ) of ~0.55, which is 2.6-fold higher than that of recombinant E. coli cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response. Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates. Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES. When cultured at 20°C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 µM CoCl2, the recombinant E. coli exhibited a specific whole cell OPH activity (U/OD600) of ~0.55, which is 2.6-fold higher than that of recombinant E. coli cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response. Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates. Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES. When cultured at 20 degree C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 mu M CoCl sub(2), the recombinant E. coli exhibited a specific whole cell OPH activity (U/OD sub(600)) of ~0.55, which is 2.6-fold higher than that of recombinant E. coli cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response. Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates. Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES. When cultured at 20°C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 µM CoCl₂, the recombinant E. coli exhibited a specific whole cell OPH activity (U/OD₆₀₀) of ~0.55, which is 2.6-fold higher than that of recombinant E. coli cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response. Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates. |
| Author | Kim, Chang Sup Song, Young Hoon Seo, Jeong Hyun Kim, Kyeoung Rok Kang, Dong Gyun |
| Author_xml | – sequence: 1 givenname: Kyeoung Rok surname: Kim fullname: Kim, Kyeoung Rok organization: Graduate School of Biochemistry, Yeungnam University – sequence: 2 givenname: Young Hoon surname: Song fullname: Song, Young Hoon organization: School of Chemical Engineering, Yeungnam University – sequence: 3 givenname: Jeong Hyun surname: Seo fullname: Seo, Jeong Hyun organization: School of Chemical Engineering, Yeungnam University – sequence: 4 givenname: Dong Gyun surname: Kang fullname: Kang, Dong Gyun organization: Department of Chemical Engineering, Pohang University of Science and Technology – sequence: 5 givenname: Chang Sup surname: Kim fullname: Kim, Chang Sup email: cskim1409@ynu.ac.kr organization: Graduate School of Biochemistry, Yeungnam University, School of Chemistry and Biochemistry, Yeungnam University |
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| CitedBy_id | crossref_primary_10_1021_acs_jafc_9b05206 crossref_primary_10_3389_fmicb_2017_00827 |
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| Keywords | GroEL-GroES organophosphorus hydrolase culture condition whole cell OPH activity |
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| Snippet | Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant
Escherichia... Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia... Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia... |
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| SubjectTerms | Analysis aryldialkylphosphatase Biocatalysts Biochemistry Biosensors Biosynthesis Biotechnology Cellular biology Chemical engineering Chemistry Chemistry and Materials Science detection limit E coli Enzymes Escherichia coli Genetic recombination Industrial and Production Engineering Organophosphates organophosphorus compounds Plasmids Proteins Research Paper storage quality Studies Temperature tetracycline toxicity 생물공학 |
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| Title | Effect of culture conditions on the whole cell activity of recombinant Escherichia coli expressing periplasmic organophosphorus hydrolase and cytosolic GroEL/ES chaperone |
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