Comparative Studies on the Susceptibility of (<i>R</i>)-2,3-Dipalmitoyloxypropylphosphonocholine (DPPnC) and Its Phospholipid Analogues to the Hydrolysis or Ethanolysis Catalyzed by Selected Lipases and Phospholipases

• Main Authors: Paweł Mituła, Czesław Wawrzeńczyk, Witold Gładkowski
• Format: Article
• Language: English
• Published: MDPI AG 2021-01-01
• Series: Catalysts
• Subjects: phosphonolipids; phospholipids; enzymatic hydrolysis; lipases; phospholipases
• Online Access: https://www.mdpi.com/2073-4344/11/1/129

Susceptibility of soybean phosphatidylcholine, 1,2-dipalmitoyl-<i>sn</i>-glycero-3-phosphocholine (DPPC) and its phosphono analogue (<i>R</i>)-2,3-dipalmitoyloxypropylphosphonocholine (DPPnC) towards selected lipases and phospholipases was compared. The ethanolysis of substrates at <i>sn</i>-1 position was carried out by lipase from <i>Mucor miehei</i> (Lipozyme^®) and lipase B from <i>Candida antarctica</i> (Novozym 435) in 95% ethanol at 30 °C, and the hydrolysis with Lecitase^TM Ultra was carried out in hexane/water at 50 °C. Hydrolysis at <i>sn</i>-2 position was carried out in isooctane/Tris-HCl/AOT system at 40 °C using phospholipase A₂ (PLA₂) from porcine pancreas and PLA₂ from bovine pancreas or 25 °C using PLA₂ from bee venom. Hydrolysis in the polar part of the studied compounds was carried out at 30 °C in acetate buffer/ethyl acetate system using phospholipase D (PLD) from <i>Streptococcus</i> sp. and PLD from white cabbage or in Tris-HCl buffer/methylene chloride system at 35 °C using PLD from <i>Streptomyces chromofuscus</i>. The results showed that the presence of C-P bond between glycerol and phosphoric acid residue in DPPnC increases the rate of enzymatic hydrolysis or ethanolysis of ester bonds at the <i>sn</i>-1 and <i>sn</i>-2 position and decreases the rate of hydrolysis in the polar head of the molecule. The most significant changes in the reaction rates were observed for reaction with PLD from <i>Streptococcus</i> sp. and PLD from <i>Streptomyces chromofuscus</i> that hydrolyzed DPPnC approximately two times slower than DPPC and soybean PC. The lower susceptibility of DPPnC towards enzymatic hydrolysis by phospholipases D gives hope for the possibility of using DPPnC-like phosphonolipids as the carriers of bioactive molecules that, instead of choline, can be bounded with diacylpropylphosphonic acids (DPPnA).