DE KREIJ, ARNO,MADRID, SUSAN MAMPUSTI,MIKKELSEN, JØRN DALGAARD,SØE, JØRN BORCH
申请号：
ES10178368
公开号：
ES2525711T3
申请日:
2004.01.15
申请国别(地区):
ES
年份:
2014
代理人:
摘要：
A method for producing a protein ester and / or protein subunit ester, a method comprising mixing an acyl donor, an acyl acceptor and water to produce a high water environment comprising 5-98% water, wherein said acyl donor is a lipid substrate selected from one or more of the group consisting of a phospholipid, a lysophospholipid, a triacylglyceride, a diglyceride, a glycolipid or a lysoglycolipid and said acyl acceptor is a protein and / or subunit of protein； and contacting the mixture with a lipid acyltransferase, such that said lipid acyltransferase catalyzes one or both of the following reactions: alcoholysis or transesterification in which the lipid acyltransferase is one that when tested using the Transferase Assay in Buffered Substrate has the minus 2% acyltransferase activity； said transferase test comprising the steps of: i) dissolving 450mg of phosphatidylcholine and 50mg of cholesterol in chloroform, evaporating to dryness in vacuo； ii) transfer 300mg of the cholesterol / phosphatidylcholine mixture to a Wheaton glass, add 15ml of 50mM HEPES buffer pH 7 and disperse the lipid in the buffer during stirring； iii) heat the substrate to 35 ° C during mixing with a magnetic stirrer and add 0.25ml of enzyme solution； iv) take 2 ml samples at 0, 5, 10, 15, 25, 40 and 60 minutes of reaction time and immediately stop the enzymatic reaction by adding 25 μl of 4M HCl to acidify the free fatty acid； v) add 3ml of chloroform and shake vigorously for 30 seconds, centrifuge and isolate 2ml of the chloroform phase, filter through a 0.45μm filter in a 10ml Dram glass tared； vi) evaporate the chloroform under a stream of nitrogen at 60 ° C, and scale the samples； vii) analyze the lipid extracted by GLC.Un método para producir un éster de proteína y/o éster de subunidad de proteína, método que comprende mezclar un donante de acilo, un aceptor de acilo y agua para producir un entorno con alto contenido en agua que comprende 5-98% de agua, en el que dicho donante de acilo e
