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Preparation of Uridine Diphosphate-N-Acetylgalactosamine from Uridine Diphosphate-N-Acetylglucosamine by Using Microbial Enzymes

机译:微生物酶从尿苷二磷酸-N-乙酰氨基葡萄糖制备尿苷二磷酸-N-乙酰半乳糖胺

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A method was developed for the large scale preparation of uridine diphosphate-N-acetylgalactosamine (UDP-GalNAc) from uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) by means of microbial enzymes. With Bacillus subtilis cell-free extract as a source of UDP-GlcNAc 4-epimerase, about 35% of the UDP-GlcNAc added was converted to UDP-GalNAc. After the residual UDP-GlcNAc was degraded to uridine triphosphate and N-acetylglucosamine-1-phosphate with a protamine-treated extract of bakers' yeast as a source of UDP-GlcNAc pyrophosphorylase, UDP-GalNAc was separated by anion-exchange column chromatography. The nucleotide was recovered by adsorption on charcoal and elution with ammoniacal ethanol. The final yield was about 100 μmol.
机译:开发了一种通过微生物酶从二磷酸尿苷-N-乙酰氨基葡萄糖(UDP-GlcNAc)大规模制备二磷酸尿苷-N-乙酰半乳糖胺(UDP-GalNAc)的方法。使用枯草芽孢杆菌无细胞提取物作为UDP-GlcNAc 4-表异构酶的来源,大约35%添加的UDP-GlcNAc被转化为UDP-GalNAc。用面包酵母处理的鱼精蛋白提取物将残余的UDP-GlcNAc降解为尿苷三磷酸和N-乙酰氨基葡萄糖-1-磷酸,作为UDP-GlcNAc焦磷酸化酶的来源,然后用阴离子交换柱色谱法分离UDP-GalNAc。通过吸附在木炭上并用氨乙醇洗脱来回收核苷酸。最终产率为约100μmol。

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