Isolation, purification and characterization of Phospholipase D from almond (Amygdalus spinosissima) from Ziarat, Balochistan

Abstract

This study was carried out for the isolation, purification, and characterization of enzyme phospholipase D (PLD) from Amygdalus spinosissima Bunge (wild thorny almond). It was observed that the crude extract showed higher values of enzyme activity 1206 U than that of purified which was found to be 915 U. The optimum time for extraction was 1.5 hours, after which the activity started decreasing. Purification of enzyme was done by hydrophobic interaction chromatography with a specific effect of calcium using Octyl Sepharose. In the absence of calcium ion, the fraction eluted from the column showed high enzymatic activity. The highest specific activity (4575 Umg -1 ), purification fold (53.2) and yield (75.8%) were obtained. Molecular weight was estimated as 62 kDa by SDS-PAGE (sodium dodecyl sulfatepolyacrylamide gel electrophoresis) analysis. Highest activity of PLD was observed at 50℃ and at 6 pH. Calcium ion stability of PLD activity was found at 40-90 mM. It is concluded from the present study that PLD has numerous industrial applications. It can be extracted and purified from A. spinosissima which is a cheap and easily available source at industrial scale.

Political and Democratic Role of The National Party in Restoring Peace and Democracy in Balochistan During the Insurgency

“…Who conducted peaceful, transparent, and impartial local government elections across the province. Among the many challenges for the NP coalition government of Balochistan after the 2013 general elections, one of the major challenges was that Balochistan was plagued by insecurity at that time (D. A. M. Baloch, 2021). How to make local elections possible in such circumstances.…”

“…In 2018, the National Party could not gain a single seat at the provincial as well as national level due to political engineering in Balochistan (D. A. M. Baloch, 2021). From 2018-2023 in these years, all the problems that were solved by the National Party in its three-year tenure have arisen again.…”

“…CPEC projects in Balochistan have come under harsh criticism from Baloch officials and activists for not doing enough for the Baloch people directly, despite their importance to the nation's economic future. It also has to reverse the ingrained pattern of purposely prioritizing Balochistan's strategic importance over the socioeconomic requirements and political rights of the Baloch people (D. A. M. Baloch, 2021). The long-running Baloch insurgency persists primarily as a result of the many Baloch people's genuine concerns.…”

“…While it's likely that some Baloch separatists have turned to foreign powers for help, a proxy conflict between competing states isn't the main cause of why large swaths of Baloch territories are still in unrest and have essentially become no-go zones for the rest of the nation. Instead of taking a more comprehensive approach that considers socioeconomic advancement and political rights, Balochistan's militarized approach to security has become a major contributor to the issue and cannot be used as a component of the province's solution (D. A. M. Baloch, 2021) and (Ahmad, 2018). The NP's remarks imply that a new window of opportunity for peace may be present.…”

“…The ground reality is that the mountains of Balochistan are both high and rocky. The hearts of the Baloch people can be reached only through the political process (D. A. M. Baloch, 2021).…”