DNA shuffling of lid removed T1 lipase and phaZ6

Due to the impressive progress of enzyme technology, our ability to create new and efficient enzyme has become possible. One of the methods to create a new enzyme is through DNA shuffling. In the present study, both enzymes T1 lipase and Poly(3-hydroxybutyrate) depolymerase (phaZ6) are hydrolases which have a similar active sites but with different substrate specificity. In this experiment, lid removed T1 lipase was shuffled with phaZ6. The lid of T1 lipase was removed so that the active site is exposed. The enzymes were fragmented by DNase and assembled through polymerase chain reaction amplification. A new enzyme was created which had a total number of 153 base pairs. It is 10X smaller in size compared to T1 lipase and phaZ6. It shows positive result in lipolytic activity screening test. However, the new enzyme shows negative result in Polyhydroxybutyrate screening test. In conclusion, a shuffled fragment has been obtained which retains the lipolytic activity but not the depolymerase activity. Therefore, the characterization and specificity of the new enzyme should be carried out.

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