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In‐Situ Co‐Cross‐Linking of Phospholipase D and Poly‐L‐Lysine for the Highly Efficient Synthesis of Phosphatidylglycerol
Author(s) -
Li Saisai,
Li Yan,
Long Nengbing,
Zhang Ruifeng
Publication year - 2019
Publication title -
journal of the american oil chemists' society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 117
eISSN - 1558-9331
pISSN - 0003-021X
DOI - 10.1002/aocs.12174
Subject(s) - phospholipase d , phosphatidylglycerol , yield (engineering) , hydrolysis , in situ , chemistry , adsorption , nuclear chemistry , materials science , phospholipid , enzyme , biochemistry , organic chemistry , membrane , phosphatidylcholine , metallurgy
Phospholipase D (PLD) was effectively immobilized on a nanocomposite support composed of ZnO nanowires and SiO 2 through in‐situ co‐cross‐linking with poly‐L‐lysine (PLL). An anionic and long‐chained bisepoxide cross‐linker was electrostatically adsorbed on the surface of ZnO nanowires prior to co‐cross‐linking. Under the optimal pH, 6.5, the loading amount of PLD reached an average of 114 mg per g support for PLL concentrations in the range of 0–0.6 mg mL −1 . Co‐cross‐linking of PLD with PLL enhanced the activity of PLD remarkably. The highest specific activity of co‐cross‐linked PLD reached 25.8 U per mg protein , which was 1.62 times as high as that of the cross‐linked PLD (15.9 U per mg protein ). (One unit of PLD activity [U] refers to the amount of PLD, which liberates 1 μmol of p‐nitrophenol per min via hydrolysis of phosphatidyl‐p‐nitrophenol at pH 8.0 and 30 °C). Under the optimized conditions, the yield of phosphatidylglycerol (PtdGro) reached 98.6% within 80 min at 45 °C and pH 6.5. The co‐cross‐linked PLD not only exhibited good resistance to pH and heat inactivation but also improved storage stability and reusability. The yield of PtdGro remained ≥85.4% after 13 repeat uses.