Analysis of lipase activity of JSF7 phage

Abstract

The lytic phage of Vibrio cholera, JSF7 has been found to have a GDSL-like lipase enzyme in its tail region. The sequencing of the organism showed that in the ORF30 of the organism there is a coding region of this protein. In this research process, we have tried to find out the effectiveness of the lipase enzyme of this phage. The experiment was conducted on trial and error, and a dye of target Rhodamine B was used throughout the process. Before the experiment, it was found that there were certain bacterial strains of vibrio (WT 324 and WT 346) that are the host of this phage. We have used these hosts to enrich our phage from time to time and used the phages of high titer around 107/ml. The research was conducted using heating techniques to extract fatty acids from the oil as our control process and then using rhodamine on the sample to find out the efficiency of Rhodamine B. Using phages on the sample we tried to find out the correlation between our obtained results. After getting acceptable results we compared our results with the negative control of the experiment which was JSF2 which had already been sequenced and no lipase enzyme coding region was found. In the last part of the research, we conducted the experiment using the process of dose dependency where we used different concentrations of phage on the sample and measured their optical density (OD) using spectrophotometer. We found that samples with the highest concentration of phage were getting the maximum result. We found that samples with the highest concentration of phage were getting the maximum OD result due to more complexing of molecules of fatty acids and rhodamine B. Thus, this was hypothesized that more phage in the sample was causing the formation of more fatty acids.