网络出版日期: 2004-07-10
基金资助
国家自然科学基金(29701005);中国科学院基础局重大项目(KJ951A150402)资助
Characterization of13C2Labeled Uridine Diphosphoglucoseof Enzymatic Synthesis by NMR
Online published: 2004-07-10
祁超 , 刘立岩 . 酶法合成13C标记尿苷二磷酸葡萄糖的核磁鉴定[J]. 中国科学院大学学报, 2004 , 21(4) : 488 -493 . DOI: 10.7523/j.issn.2095-6134.2004.4.010
The authors set up enzymatic method of one step to synthesize UDPG in much higher yields, which iseconomical and cheap. In enzymatic synthesis, UTP is used to replace ATP, and repetitive system of UTP is setup. Repetitive addition and recovery of enzyme are used to take advantage of enzyme. UDP-[4-13C]-glucose are firstsynthesized on 0.5 g scale by this method. The overall yield is 69.7 %. NMR spectra of 13C-labelled UDPGmakecertain the assignment of C-4 of the glucose residue and suggest that part of the results in the literatures about theassignment of UDPG are wrong
Key words： UDPG; enzymatic synthesis; enzymatic method of one step; NMR
[ 1 ] Calib E, Leloir L G. The biosynthsis of trehalose phosphate. J. Biol. Chem., 1958, 231 : 259 —275.
[ 2 ] Carpita N C, Delmer D P. Concentration and metabolic turnover of UDP2glucose in developing cotton fibers. J. Biol. Chem., 1981, 25 : 308 —315.
[ 3 ] Feingold D S, Baeber GA. In methods in plant biochemistry. In : Dey P M, Harbone J B, eds. Vol. 2. New York :Academic press, 1990.39.
[ 4 ] Moffatt J G, Khorana H G. Nucleoside polyphosphates, VIII. New and improved synthesis of uridine diphosphate glucose and flavin adeninedinucleotide using nucleoside25′2 phosphoramides. J. Am. Chem. Soc., 1958, 80 :3756 —3761.
[ 5 ] Moffatt J G. Sugar nucleotide synthesis by the phosphoromorpholidate procedure. In : Methods Enzymol.. Ⅷ. 1966. 137 —145.
[ 6 ] Hanessian S, Lu P P, Ishida H. One2step, stereocontrolled synthesis of glycosyl212phosphates, uridine25′2 diphosphogalactose, anduridine25′2diphosphoglucose from unprotected glycosyl donors. J. Am. Chem. Soc., 1998, 120 :13296 —13300.
[ 7 ] Zervoson A, Stein A, Adrian H, Elling L. Combined enzymatic synthesis of nucleotide ( deoxy) sugars from sucrose and nucleosidemonophosphates. Tetrahedron, 1996, 52 :2395 —2404.
[ 8 ] 祁 超. Apocynaceae 系细胞尿苷二磷酸葡萄糖焦磷酸化酶的纯化和表征. 中国科学院研究生院学报,2004,21 (3) :345 —351.
[ 9 ] Munch2Peterson A, Kalckar H M, Cutolo E, Smith E E B. Nature, 1953, 172 : 1036 —1037.
[10 ] Tan A W H. A simplified method for the preparation pure UDP2[4213C]2glucose. Biochem. Biophys. Acta., 1979, 582 :543 —547
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