Page 41 - חוברת תזות הנדסה ירוקה 2023

In silico catalytic property analysis of a novel prenyltransferase from Pistacia palaestina
Damian Alejandro Kischinovsky1; damiankischinovsky@gmail.com
Dr. Yoram Shotland1, Prof. Efraim Lewinsohn2
1Sami Shamoon College of Engineering, Beer-Sheva, Israel.
2Newe Ya’ar Research Center, Agricultural Research Organization, Volcani Center, Ramat Yishay, Israel.
Terpenes are secondary metabolites that fulfill different essential functions in the life of plants and other organisms and are important in industry as a source of perfumes, turpentine, essential oils, and pharmaceuticals. Plants are the largest source of terpenes. However, their extraction may has harmful environmental effects, such as: deforestation; the excessive use of land and water for the cultivation of certain species; and/or the pollution of water, land, and air by toxic residues from synthetic industrial processes. At an economic level, the high maintenance costs of the infrastructure necessary for the extraction and processing of terpenes and their derivatives are not justified by the low production yield, thus increasing the price of the end product and making these processes unprofitable.
As such, it is crucial to provide biotechnological tools that enable transgenic microorganisms to produce terpenes. To that end, it is imperative to understand the processes, enzymes,
and genes involved in terpene biosynthesis. Among the most important enzymes in this process are the prenyltransferases )PTs( that, together with other enzymes, synthesize the wide variety of terpenes used in our industry.
The infestation of plants by aphids is a highly relevant topic, and the study of plant-aphid interactions has provided several valuable findings. Baizongia pistacia )the horn gall aphid( is an aphid that, by attacking the Pistacia palaestina tree, induces the formation of galls for its proliferation. A study of secondary metabolite levels in Pistacia palaestina revealed that monoterpene levels in galls were higher than in healthy leaves. This phenomenon was accompanied by an increase in the expression of certain prenyltransferases, especially a small geranyl diphosphate )GPP( synthase subunit called PpPRT2, suggesting that this protein may have catalytic properties and is involved in monoterpene accumulation.
The objective of this work was to characterize the PpPRT2 protein and its possible involvement in the synthesis of GPP, the precursor of monoterpenes. Our study, based on in silico methods, revealed that the PpPRT2 protein possesses a catalytic site capable of interacting with DMAPP and IPP substrates through divalent magnesium ions. The binding affinity values were consistent with values detected in a large subunit in Mentha piperita GPP synthase, an active catalytic prenyltransferase.
The mutation of specific residues from PpPRT2 revealed that this catalytic property may
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