Obtain gene of MlrA
MlrA is a name given to the enzyme that breakdown microcystin-LR. We initiated our design by searching for alternative MlrA gene sequences in the National Center for biotechnology information. Cluster analysis was performed by the software MAGE to compare those genes. We were eventually able to select the optimal genes of MlrA from two species Novosphingobium sp. THN1 and Sphingopyxis sp. ACM-3962. We named them as MlrA2 and MlrA3 respectively during our study.
Examine the Microcystin degradation
HPLC separate solvents according to their polarity. We mixed the microcystin-LR with the MlrA at different proportion and left them to react for a specific amount of time, then injected the reacted mixture to the HPLC device. After tuning the machine to the right way and programming the progress.we could see the amount of micocystin-LR degraded to find out how efficient does the MlrA work.
Immobilizing the MlrA
To not waste any of MlrA we made with painstaking effort, immobilizing it and reusing it is the best way to reduce cost of production. We plan to use alginate beads to seal the MlrA. The MlrA is mixed with the sodium alginate solution, and dropped to the calcium chloride solution. When the two solution meet, they instantly react to form jelly. Thus, MlrA can be hold tight in the beads after solidification. We can fill a column by those beads and when the water with microcystin-LR passes through the column, it can be purified.