Abstract: Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage in 4oC without vacuum. Polyethylene glycol (PEG) pre-treated freeze dry cells and broth pre-treated freeze dry cells after freeze-dry recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introduce freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 56.78% and 17.91%. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks were 26.35% and 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been growth in agar. Result from this study may be beneficial and useful as initial reference before commercialize freeze-dried P. putida.
Abstract: Microbes have been used to solve environmental
problems for many years. The role of microorganism to sequester,
precipitate or alter the oxidation state of various heavy metals has
been extensively studied. Treatment using microorganism interacts
with toxic metal are very diverse. The purpose of this research is to
remove the mercury using Pseudomonas putida (P. putida), pure
culture ATTC 49128 at optimum growth parameters such as
techniques of culture, acclimatization time and speed of incubator
shaker. Thus, in this study, the optimum growth parameters of P.
putida were obtained to achieve the maximum of mercury removal.
Based on the optimum parameters of P. putida for specific growth
rate, the removal of two different mercury concentration, 1 ppm and
4 ppm were studied. From mercury nitrate solution, a mercuryresistant
bacterial strain which is able to reduce from ionic mercury
to metallic mercury was used to reduce ionic mercury. The overall
levels of mercury removal in this study were between 80% and 89%.
The information obtained in this study is of fundamental for
understanding of the survival of P. putida ATTC 49128 in mercury
solution. Thus, microbial mercury removal is a potential
bioremediation for wastewater especially in petrochemical industries
in Malaysia.