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Optimum Conditions of Freezing Lyophilization and Bioluminescence Activity Recovery for Environmental Applications Using a Recombinant Strain  

Ko Kyung-Seok (Geological & Environmental Hazards Division, Korea Institute of Geoscience & Mineral Resources (KIGAM))
Kim Myung-Hee (School of Construction and Environmental Engineering, Yeungnam University)
Kong In-Chul (School of Construction and Environmental Engineering, Yeungnam University)
Publication Information
Journal of Soil and Groundwater Environment / v.11, no.5, 2006 , pp. 43-50 More about this Journal
Abstract
Bioreporter bacteria, such as recombinant bioluminescent bacteria, have been used for the detection of specific compounds in complex environmental media. In this study, optimum conditions for the preparation and application of deep-freezed and Iyophilized recombinant bioluminescent strain KG1206 were investigated for the future application on contaminated environmental sites. Genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, contains TOL plasmid and the plasmid inserted $P_{m}$, promoter on the upper part of lux gone in vector pUCD615, and m-toluate and benzoate are considered direct inducers for bioluminescence. Optimum conditions determined for the preparation and application of the deep-freezed and lyophilized strain were followings: cryoprotective agent (24% sucrose), lyophilization time (12 hrs), strain concentration ($OD_{600}=0.6$), reconstitution for freezed strain (quick reconstitution at $35^{\circ}C$), reconstitution for lyophilized strain ($3{\sim}6$ hrs exposure on LB medium), carrying conditions (keep at $20^{\circ}C$ after reconstitution). These results demonstrate the feasibility of deep-freezed or lyophilized state of genetically engineered bioluminescent strain for environmental usage.
Keywords
Genetically engineered microorganism(GEM); Bioluminescence; Deep-freeze; Lyophilization; TOL-plasmid;
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