Lipozyme calb l

Phosphatidylcholine (PC>98%), phosphatidyl ethanolamine (PE>80%) and phosphatidylserine (PS>50%) was purchased from Merya’s lecithin co., ltd. (Beijing, China). Refined DHA-rich algal oil from Schizochytrium sp. (DHA ≥60%) was kindly provided by Qingdao Xunon Bioengineering Co., Ltd. (Qingdao, China). DHA and DHA-ME were prepared from algal oil. The 37 kinds of fatty acid methyl ester standards were purchased from Sigma-Aldrich Co., Ltd. (Norcross, GA). Lipozyme^®CALB L (from Candida antarctica, CALB), Lipozyme^® TL 100 L (from Thermomyces lanuginosus, TL100), Novocor^®AD L (from Candida antarctica, ADL) Novozym^®51,032 (from Aspergillus oryzae, 51,032), Resinase^®HT (from Aspergillus oryzae, HT), Lecitase^® Ultra (from Thermomyces lanuginosus/Fusarium oxysporum, PLA₁), Novozym^®435 (from Candida antarctica, 435), and Lipozyme^®TL IM (from Thermomyces lanuginosus, IM) were purchased from Novozymes Biotechnology Co., LTD. (Tianjin, China). The thin-layer chromatography plates coated with silica gel G were purchased from Qingdao Ocean Chemical Co., Ltd. (Qingdao, China). Other chemicals and solvents applied were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China) with chromatographic and analytical grade.

Choline chloride > 98%, betaine 98%, D(+)-glucose 99.5%, D(-)-fructose, polyethyleneimine, branched (PEI), ethyl-4-hydroxy-3-methoxycinnamate (ethyl ferulate) 4-nitrophenyl butyrate (pNPB) > 98%, and p-nitrophenol (pNP) were obtained from Sigma-Aldrich, St. Louis, MI, USA. Choline dihydrogen phosphate >98% was bought from IOLITEC, (Heilbronn, Germany), ethylammonium chloride from Merck (Sigma-Aldrich, USA), urea > 99% from Fluka (Sigma-Aldrich, USA), glycerol 0.5% max water, and Glutaraldehyde 25% solution from Fisher Scientific (Hampton, NH, USA), ethylene glycol from AppliChem (Darmstadt, Germany), 1,2-butanediol > 98% from TCI (Japan), 1,2-propanediol 99% from Acros Organics (Waltham, MA, USA), and sodium hydroxide > 98% from Panreac (Barcelona, Spain). The HPLC solvents acetonitrile, methanol, and water were of HPLC grade and the water used was double distilled. The lipases from Candida antarctica Lipozyme CALB L (liquid) and Novozym 435 (immobilized) were kindly offered by Novozymes (Bagsværd, Denmark). PLA filament designed for use with FDM 3D printers (PrimaValue™) was purchased from Prima Creator (Malmö, Sweden). The 3D printer used was an Ender 5 from Creality 3D (Shenzhen, China).

Lipozyme® CALB L, lipase B of C. antarctica (CALB, EC 3.1.1.3, 19.1 U/mg protein; 7.50 mg protein/ml) was kindly provided by Novozymes (Bagsvaerd, Denmark).