Preparative 2'-Reduction of ATP Catalyzed by Ribonucleotide Reductase Purified by Liquid-Liquid Extraction

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概要

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• Recombinant Lactobacillus leichmannii ribonucleosidetriphosphate reductase (RTPR, E.C.1.17.4.2) constitutively expressed by E. coli HB101 pSQUIRE has been purified from sonicated cell material in a one-step procedure by PEG 4000 (16% (w/w))/phosphate (7% (w/w)) liquid-liquid extraction. A high yield of 75.1% RTPR in the top phase and a partitioning of 8.5 : 1 between total RTPR activity in to and bottom phase were obtained in this preparative system. The RTPR-containing top phase was used to reduce ATP in the 2'-position on a gram scale with high final conversion and yield proving the ribonucleotide reductase approach feasible for the preparative synthesis of 2'-deoxyribonucleotides. High concentrations of sodium acetate in the reaction served to substitute for allosteric effectors of RTPR. 1,4-Dithio-DL-threitol was used as an artificial reducing agent for RTPR.

• 社団法人日本農芸化学会の論文
• 2000-09-23

著者

• Ghisalba Oreste

  Novartis Pharma Ag

• BRUNELLA Andre

  Outcomes International

• ABRANTES Miguel

  Logoplaste SA

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• Preparative 2'-Reduction of ATP Catalyzed by Ribonucleotide Reductase Purified by Liquid-Liquid Extraction
• Chemo-enzymatic Synthesis of both Enantiomers of myo-Inositol 1,3,4,5-tetrakisphosphate
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