Optimization of a PCR Method for Detection of Lipase Gene in Bacillus subtilis

Abstract

Lipases catalyze hydrolysis and synthesis of triacylglycerols. These enzymes,especially the microbial ones, have wide commercial and industrial usage. <em><span style="color: #231f20; font-family: Times New Roman;"><span style="color: #231f20; font-family: Times New Roman;">Bacillus </span></span></em>subtilis <span style="color: #231f20; font-family: Times New Roman;"><span style="color: #231f20; font-family: Times New Roman;">lipase has interesting properties such as small size and tolerance to basic pH;</span></span> therefore, developing techniques for its recognition and isolation is the focus of research in many laboratories. In the present study, two factors i.e. MgCl<span style="color: #231f20; font-family: Times New Roman; font-size: xx-small;"><span style="color: #231f20; font-family: Times New Roman; font-size: xx-small;"><span style="color: #231f20; font-family: Times New Roman; font-size: xx-small;">2 </span></span></span>concentration and annealing temperature were manipulated for the optimization of the polymerase chain reaction (PCR) condition. As for the MgCl <span style="color: #231f20; font-family: Times New Roman; font-size: xx-small;"><span style="color: #231f20; font-family: Times New Roman; font-size: xx-small;"><span style="color: #231f20; font-family: Times New Roman; font-size: xx-small;">2 </span></span></span><span style="color: #231f20; font-family: Times New Roman;"><span style="color: #231f20; font-family: Times New Roman;">concentration, 2.5 </span></span>mM produced the best results when the annealing temperature was kept at 55 <span style="line-height: 115%; font-family: 'Times New Roman','serif'; font-size: 12pt; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">°C</span><span style="color: #231f20; font-family: Times New Roman;"><span style="color: #231f20; font-family: Times New Roman;">. </span></span>Regarding the annealing temperatures, the best amplification was obtained at 63.9 <span style="line-height: 115%; font-family: 'Times New Roman','serif'; font-size: 12pt; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">°C</span>. <span style="color: #231f20; font-family: Times New Roman;"><span style="color: #231f20; font-family: Times New Roman;"> It can be concluded that the PCR conditions for the detection of lipase gene in </span></span>Bacillus subtilis <span style="color: #231f20; font-family: Times New Roman;"><span style="color: #231f20; font-family: Times New Roman;">has been optimized and can be used for the screening and isolation </span></span>of these bacaterial strains.