Contact
    Start Publications Deciphering the recombinant thermostable ...
    ←  Publication Finder
    KSV NIMA

    Deciphering the recombinant thermostable phosphatidylcholine-specific phospholipase C activity from Bacillus thuringiensis: Biochemical and interfacial properties

    Year: 2021

    Journal: Colloid Surf. A-Physicochem. Eng. Asp., Volume 630, DEC 5

    Authors: Eddehech, Ahlem; Smichi, Nabil; Violot, Sebastien; Bettler, Emmanuel; Brizuela, Leyre; Noiriel, Alexandre; Abousalham, Abdelkarim; Zarai, Zied

    Organizations: Tunisian Ministry of Higher Education and Scientific Research, Tunisia [18PJEC08-04]; PHC-Maghreb program of the French Ministry of Foreign Affairs; Ministry of Higher Education, Research and Innovation, France [43791TM, PHC: 01MAG20]

    Keywords: B. thuringiensis IL14; PC-PLC; Thermostability; Substrate specificity; Interfacial properties; Cytotoxic properties

    A novel alkaline thermostable phosphatidylcholine-specific phospholipase C (PC-PLCBt) was expressed in E. coli system. Recombinant PC-PLCBt (rPC-PLCBt) activity and thermostability were shown to be significantly depen-dent on the Zn2+. The maximum rPC-PLCBt catalytic activity was found to be 1372 U mg(-1) in the presence of 0.1 mM Zn2+ and at 60 degrees C using an Egg PC as substrate. The interfacial kinetic data show that nPC-PLCBt and rPC-PLCBt display similar substrate specificity on various phospholipid monolayers. The maximal rPC-PLCBt activities were recorded, at decreasing order, on 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE), 1,2-diacyl-sn-phosphoglycerol (PG), and 1,2-diacyl-sn-phosphoserine (PS) monolayers at interfacial surface pressures of 15, 25, 20, and 25 mN m(-1), respectively. Such important penetrating power could be exploited for pharmacological purposes. The highest activities were recorded on the DLPC monolayer and shown to be 121.61 and 40.13 mmol cm(-2) min(-1) M-1 for native and recombinant PC-PLCBt, respectively. Interestingly, compared to all known Bacillus PLCs, both PC-PLCBt forms showed an exclusive capacity to hydrolyze the PG film with a more pronounced rate of hydrolysis for the native form with a specific activity of 58.29 mmol cm(-2) min(-1) M-1. Therefore, the high enzyme level production of about 14 mg L-1, the thermostability as well as the broad phospholipid specificity of PC-PLCBt represent great potential in the crude oil refining industry.
    View publication