Massou S, Puech V, Talmont F, Demange P, Lindley N D, Tropis M, Milon A
IPBS-CNRS, Toulouse, France.
J Biomol NMR. 1999 Jul;14(3):231-9. doi: 10.1023/a:1008371118772.
Methylotrophic yeast has previously been shown to be an excellent system for the cost-effective production of perdeuterated biomass and for the heterologous expression of membrane receptors. A protocol for the expression of 85% deuterated, functional human mu-opiate receptor was established. For partially deuterated biomass, deuteration level and distribution were determined for fatty acids, amino acids and carbohydrates. It was shown that prior to biosynthesis of lipids and amino acids (and of carbohydrates, to a lower extent), exchange occurs between water and methanol hydrogen atoms, so that 80%-90% randomly deuterated biomass and over-expressed proteins may be obtained using only deuterated water.
甲基营养型酵母此前已被证明是一种用于经济高效地生产全氘代生物质以及用于膜受体异源表达的出色系统。已建立了一种表达85%氘代的功能性人μ-阿片受体的方案。对于部分氘代生物质,测定了脂肪酸、氨基酸和碳水化合物的氘代水平和分布。结果表明,在脂质和氨基酸(以及程度较低的碳水化合物)生物合成之前,水和甲醇氢原子之间会发生交换,因此仅使用氘代水就可以获得80%-90%随机氘代的生物质和过表达的蛋白质。