PRACTICAL SYNTHESIS OF 5-PHOSPHO-D-RIBOSYL ALPHA-1-PYROPHOSPHATE (PRPP) - ENZYMATIC ROUTES FROM RIBOSE 5-PHOSPHATE OR RIBOSE
PRACTICAL SYNTHESIS OF 5-PHOSPHO-D-RIBOSYL ALPHA-1-PYROPHOSPHATE (PRPP) - ENZYMATIC ROUTES FROM RIBOSE 5-PHOSPHATE OR RIBOSE
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DOI：
10.1021/ja00363a037
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发表时间：
1983-01-01
影响因子：
15
通讯作者：
WHITESIDES, GM
中科院分区：
文献类型：
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作者：
GROSS, A;ABRIL, O;WHITESIDES, GM
本文描述了5-磷酸-D-核糖基α- 1-在75-mmol规模上的焦磷酸盐（PRPP）。反应使用PAN固定化的PRPP合成酶作为催化剂，原位ATP-辅因子再生。在一个程序中，纯r-5-P被用作起始材料;在第二个程序中，r-5-P通过核糖激酶催化的D-核糖磷酸化合成并原位使用。通过UMP的酶促合成证明了PRPP用作制备核苷酸的起始材料的潜力。本文还介绍了几种制备r-5-P的方法：AMP的酸催化水解，通过消化RNA获得的粗单肽混合物的酸催化水解，D-核糖的化学合成和D-核糖的核糖激酶催化合成。描述了分离PRPP合成酶（来自鼠伤寒沙门氏菌）和核糖激酶（来自植物乳杆菌）以及将这些酶固定在PAN中的程序。
This paper describes enzymatic syntheses of 5-phospho-D-ribosyl .alpha.-1-pyrophosphate (PRPP) on a 75-mmol scale. The reactions used PAN-immobilized PRPP synthetase as catalyst with in situ ATP-cofactor regeneration. In one procedure pure r-5-P was used as a starting material; in a second, r-5-P was synthesized by ribokinase-catalyzed phosphorylation of D-ribose and used in situ. The potential for use of PRPP as a starting material for the preparation of nucleotides was demonstrated by an enzymatic synthesis of UMP. This paper also describes several methods for the preparation of r-5-P; acid-catalyzed hydrolysis of AMP, acid-catalyzed hydrolysis of a crude mononucleotide mixture obtained by digestion of RNA, chemical synthesis from D-ribose and ribokinase-catalyzed synthesis from D-ribose. Procedures are described for the isolation of PRPP synthetase (from Salmonella typhimurium) and ribokinase (from Lactobacillus plantarum) and for the immobilization of these enzymes in PAN.