Hanko Valoran P, Rohrer Jeffrey S
Dionex Corporation, Sunnyvale, CA 94088-3603, USA.
Anal Biochem. 2002 Sep 15;308(2):204-9. doi: 10.1016/s0003-2697(02)00215-4.
Here we present a new method to rapidly quantify tryptophan (Trp) in proteins, animal feed (Mehaden fishmeal), cell cultures, and fermentation broths. Trp is separated from common amino acids by anion-exchange chromatography in 12min and directly detected by integrated pulsed amperometry. The estimated lower detection limit for this method is 1pmol. Alkaline (4M NaOH) hydrolysates can be directly injected, and therefore we used this method to determine the optimum alkaline hydrolysis conditions for the release of Trp from a model protein, bovine serum albumin (BSA). This method accurately determined the Trp content of BSA and fishmeal. High levels of glucose (2%, w/w) do not interfere with the chromatography or decrease recovery of Trp. We used this method to monitor free Trp during an Escherichia coli fermentation.
在此，我们提出了一种新方法，可快速定量测定蛋白质、动物饲料（鲱鱼粉）、细胞培养物和发酵液中的色氨酸（Trp）。色氨酸通过阴离子交换色谱法在12分钟内与常见氨基酸分离，并通过集成脉冲安培法直接检测。该方法的估计检测下限为1皮摩尔。碱性（4M氢氧化钠）水解产物可直接进样，因此我们使用该方法确定了从模型蛋白牛血清白蛋白（BSA）中释放色氨酸的最佳碱性水解条件。该方法准确测定了BSA和鱼粉中的色氨酸含量。高浓度葡萄糖（2%，w/w）不会干扰色谱分析，也不会降低色氨酸的回收率。我们使用该方法监测大肠杆菌发酵过程中的游离色氨酸。
