Nakazawa Yozo
Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, Abashiri, Hokkaido, Japan.
Methods Mol Biol. 2012;861:179-200. doi: 10.1007/978-1-61779-600-5_12.
The transphosphatidylation catalytic ability of phospholipase D (PLD, EC 3.1.4.4) is a powerful biochemical tool for the acquisition of rare phospholipids (PLs), e.g., phosphatidylglycerol (PG) and phosphatidylserine (PS), and artificial phospholipids, which do not occur in nature. Specifically, actinomycete PLD recognizes not only the alcohols (i.e., glycerol or serine) corresponding to the polar head groups of natural PLs, but also secondary alcohols, aromatic alcohols, saccharides, N-heterocyclic alcohols, and vitamins as acceptors. Therefore, actinomycete PLD is a valuable enzyme in food, cosmetics, fine chemical and pharmaceutical industries. Here, we describe a protocol for the screening for PLD-producing microorganisms, several PLD assays and methods of PLD production-purification and the strategy of cloning of the Streptomyces PLD gene.
磷脂酶D（PLD，EC 3.1.4.4）的转磷脂酰化催化能力是获取稀有磷脂（PLs），如磷脂酰甘油（PG）和磷脂酰丝氨酸（PS）以及自然界中不存在的人工磷脂的强大生化工具。具体而言，放线菌PLD不仅能识别与天然PLs极性头部基团相对应的醇类（即甘油或丝氨酸），还能识别仲醇、芳香醇、糖类、N-杂环醇和维生素作为受体。因此，放线菌PLD在食品、化妆品、精细化工和制药行业中是一种有价值的酶。在此，我们描述了一种筛选产PLD微生物的方案、几种PLD测定方法以及PLD生产-纯化方法和链霉菌PLD基因的克隆策略。
