Development of Methods for Immobilization and Usages of Immobilized Recombinant Bioluminescent Strain, Pseudomonas Putida Mt-2 KG1206, Stored by Deep-FreezingKim M1 and Kong IC2*
- Corresponding Author:
- In Chul Kong
Department of Environmental Engineering
Kyungbuk 38541, Korea
Fax: +82- 53-810-4624
E-mail: [email protected]
Received date: September 05, 2015; Accepted date: November 20, 2015; Published date: November 27, 2015
Citation: Kim M, Kong IC (2015) Development of Methods for Immobilization and Usages of Immobilized Recombinant Bioluminescent Strain, Pseudomonas putida Mt-2 KG1206, Stored by Deep-Freezing. J Biotechnol Biomater 5:208. doi:10.4172/2155-952X.1000208
Copyright: © 2015 Kim M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Deep-freezing process can be used practically for environmental monitoring. However, for the organism to be useful in environmental applications, the strain of bacteria exhibits a high intensity of activity following reconstitution after deep-freezing. An overall protocol for immobilization and reconstitution of recombinant bioluminescent bacteria, preserved by deep-freezing was investigated for future biomonitoring. Tested strain KG1206 has ability to produce bioluminescence in the presence of toluene analogs and those intermediates. Immobilization using glass beads with subcultured cells showed better results than other conditions, such as alginate beads or centrifuged cell pellets. Beads number, thawing time, and amount of reconstitution solution influence on bioluminescence activity. Among tested conditions, followings were appeared as optimum conditions: working volume 15 mL of serum vials, approximately 20-30 glass beads per vial, 3 h thawing time, and 2-5 mL reconstitution solution. Potassium nitrate (KNO3) greatly stimulates the low bioluminescence activity of immobilized strain, preserved by deep freezing, in the range of 2.5 to 21 times of untreated one.