Clarke A J
Department of Microbiology, University of Guelph, Ontario, Canada.
Anal Biochem. 1993 Aug 1;212(2):344-50. doi: 10.1006/abio.1993.1339.
A high-performance anion-exchange chromatography method with pulsed-amperometric detection has been developed for the simultaneous analysis of both amino acids and amino sugars and applied to the compositional analysis of peptidoglycan hydrolysates. Chromatography of the acid hydrolysis products was performed on a CarboPac PA-1 anion-exchange column, with pulsed-amperometric detection. Complete resolution of the two amino sugars (glucosamine and muramic acid) and eight of the nine amino acids (Ala, diaminobutyric acid, diaminopimelic acid, Glu, Gly, homoserine, Lys, Orn, and Ser) known to occur in various peptidoglycans was achieved within 70 min. Only homoserine and glycine (retention times 26.8 and 26.9 min, respectively) were not resolved by this procedure, but the simultaneous occurrence of these two amino acids in peptidoglycan is extremely rare. Reproducibility of the separations was shown to be very high and detection limits exceeded 10 pmol for glucosamine. This convenient and simple analysis was applied to the quantitation of many crude peptidoglycan samples isolated from the species of the Proteeae (Proteus, Providencia, and Morganella) for the determination of the extent of peptidoglycan O-acetylation.
已开发出一种带脉冲安培检测的高效阴离子交换色谱法，用于同时分析氨基酸和氨基糖，并应用于肽聚糖水解产物的组成分析。酸水解产物的色谱分析在CarboPac PA-1阴离子交换柱上进行，并采用脉冲安培检测。在70分钟内实现了两种氨基糖（葡萄糖胺和胞壁酸）以及已知存在于各种肽聚糖中的九种氨基酸中的八种（丙氨酸、二氨基丁酸、二氨基庚二酸、谷氨酸、甘氨酸、高丝氨酸、赖氨酸、鸟氨酸和丝氨酸）的完全分离。只有高丝氨酸和甘氨酸（保留时间分别为26.8和26.9分钟）未通过此方法分离，但这两种氨基酸同时存在于肽聚糖中的情况极为罕见。分离的重现性非常高，葡萄糖胺的检测限超过10皮摩尔。这种简便的分析方法应用于从变形杆菌属（变形杆菌、普罗威登斯菌和摩根菌）分离的许多粗肽聚糖样品的定量分析，以确定肽聚糖O-乙酰化的程度。
