A simple and cost-effective protocol is presented for expression of perdeuterated, Ile/Leu/Val ¹H/¹³C methyl protonated proteins from 100 ml cultures in M9 ++ /D₂O medium induced at high (OD₆₀₀ ~ 10) cell density in shaker flasks. This protocol, which is an extension of our previous protocols for expression of ²H/¹⁵N/¹³C and ¹H/¹³C labeled proteins, yields comparable quantities of protein from 100 ml cell culture to those obtained using a conventional 1 L culture with M9/D₂O medium, while using three-fold less α-ketoisovaleric (1,2,3,4-¹³C₄; 3,4′,4′,4′-d₄) and α-ketobutyric (¹³C₄; 3,3-d₂) acid precursors.

提出了一种简单且具有成本效益的方案，用于在高 (OD ₆₀₀  ~ 10) 细胞诱导的 M9 ++ /D ₂ O 培养基中从 100 ml 培养物中表达过氘化的 Ile/Leu/Val ¹ H/ ¹³ C 甲基质子化蛋白摇瓶中的密度。该方案是我们之前表达² H/ ¹⁵ N/ ¹³ C 和¹ H/ ¹³ C 标记蛋白的方案的扩展，从 100 ml 细胞培养物中产生的蛋白质数量与使用传统 1 L 培养物获得的蛋白质数量相当使用 M9/D ₂ O 培养基，同时使用少三倍的 α-酮异戊酸 (1,2,3,4- ¹³ C ₄; 3,4',4',4'-d ₄ ) 和 α-酮丁酸 ( ¹³ C ₄ ; 3,3-d ₂ ) 酸前体。