Effect of culture conditions on the whole cell activity of recombinant Escherichia coli expressing periplasmic organophosphorus hydrolase and cytosolic GroEL/ES chaperone

• Published in: Biotechnology and bioprocess engineering Vol. 21; no. 4; pp. 502 - 507
• Main Authors: Kim, Kyeoung Rok, Song, Young Hoon, Seo, Jeong Hyun, Kang, Dong Gyun, Kim, Chang Sup
• Format: Journal Article
• Language: English
• Published: Seoul The Korean Society for Biotechnology and Bioengineering 01.08.2016; Springer Nature B.V; 한국생물공학회
• Subjects: 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; GroEL-GroES; organophosphorus hydrolase; culture condition; whole cell OPH activity
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-016-0342-y

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.