谷氨酸棒杆菌代谢产物尿苷二磷酸葡萄糖与尿苷二磷酸半乳糖的高效液相色谱测定方法研究.pdfVIP

L050 氨酸棒杆菌代谢产物尿苷二磷酸葡萄糖与尿苷二磷酸 半乳糖的高效液相色谱测定方法 小杰 何宁 李清彪 邓旭 卢英华 姚传义 （厦门大学化学工程与生物工程系，福建厦门 361005 ） 要 采用Zorbax NH 色谱柱 id.4.6 mm ×250 mm，5 μm），以0.125 mol/L KH PO 缓冲液(pH 3.6) 2 2 4 和乙腈为流动相分离谷氨酸棒杆菌胞内代谢产物尿苷二磷酸葡萄糖和尿苷二磷酸半乳糖，得到较好 分离效果。UDP-葡萄糖和UDP-半乳糖的停留时间分别为 12.1 min 和 12.7 min。精确度和回收率实 验证明，此高效液相法对于测定UDP-葡萄糖和UDP-半乳糖是一种便捷可靠的方法。初步探讨了谷氨 酸棒杆菌代谢过程中胞内UDP-葡萄糖和UDP-半乳糖的合成与絮凝剂活性之间的相关性。 关键词 高效液相色谱 尿苷二磷酸葡萄糖 尿苷二磷酸半乳糖 谷氨酸棒杆菌 絮凝剂 SEPARATION OF INTRACELLULAR INTERMEDIATES URIDINE DIPHOSPHOGLUCOSE AND URIDINE DIPHOSPHOGALACTOSE OF CORYNEBACTERIUM GLUTAMICUM BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WU Xiaojie HE Ning LI Qingbiao DENG Xu LU Yinghua YAO Chuanyi (Depa rtment of Chemical and Biochemical Engineering,X iamen University, X iamen 361005,Fuj ian,P.R. China) Abstract Uridine diphosphoglucose (UDP-Glc) and Uridine diphosphogalactose (UDP-Gla) in the cell free extracts of Cory nebacterium glutamicum CCTCC M111 were successfully separated using Zorbax NH2 column (id.4.6 mm ×250 mm，5 μm) with 0.125 mol/L potassium phosphate buffer(pH 3.6)-acetonitrile. The retention time of UDP-Glucose and UDP-Glactose were 12.1 minutes and 12.7 minutes respectively. With the calibration line of UDP-Glc and UDP-Gal，recovery rate and precision, HPLC was proven to be a suitable way for the assay of UDP-Gal and UDP-Glc in the cells of Cory nebacterium glutamicum CCTCC M 111. Relationship between the content of UDP-Gal, UDP-Glc and bioflocculant production was also discussed in this paper. Keywords HPLC, uridine diphosphoglucose, uridine diphophogalatose, Cory nebacterium 1 glutamicum, bioflocculant 引 言 尿苷二磷酸葡萄糖 UDP-葡萄糖，简称 UDP