CDP-Ethanolamine and CDP-Choline: one-pot synthesis and 31P NMR study

Ghezal, Salma; Thomasson, Maggie S.; Lefebvre-Tournier, Isabelle; Philouze, Christian; Macnaughtan, Megan A.; Roy, Béatrice

doi:10.1016/j.tetlet.2014.07.076

Cited by 8 publications

(11 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The predicted initially formed products for Cj1416 with MnCTP and the substrates in Scheme are shown in Scheme . The Cj1416-catalyzed reaction products, cytidine diphosphoramidate ( 3 ), CDP- l -glutamine ( 2 ), CDP ( 16 ), and CDP-ethanolamine ( 20 ) using phosphoramidate ( 5 ), l -glutamine-P ( 1 ), phosphate ( 6 ), and ethanolamine-P ( 10 ) are consistent with their previously published 31 P NMR spectra (Figures and ). , The chemical shifts for four other reaction products, O -methyl CDP ( 17 ), CMP methyl phosphonate ( 18 ), CDP- l -serine ( 25 ), and CDP-serinol ( 24 ), are also consistent with their predicted structures (Figure S1). The remaining four substrates, 3-phospho- d -glycerate ( 11 ), ( R / S )-glycerol-1-phosphate ( 12 ), glycerol 2-phosphate ( 13 ), and arsenate ( 9 ), all exhibited 31 P NMR spectra that were inconsistent with the predicted products.…”

Section: Resultssupporting

confidence: 86%

“…Methyl phosphate (7) and methyl phosphonate (8) were both assayed as possible substrates, since they are each similar in size and shape to phosphoramidate (5). Cj1416 catalyzed the formation of the O-methyl ester of CDP (17) and CMP methyl phosphonate (18). In total, 35 different compounds were assayed for catalytic activity with Cj1416 and MnCTP (Scheme S1).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…If O-labeled phosphate is used as an alternate substrate, Cj1416 can be used to form β-[ 18 O 4 ]-CDP, which can subsequently be used to synthesize β-[ 18 O 4 ]-CTP. Typically, β-[ 18 O 4 ]-CDP is made using morpholidate chemistry in dry DMSO with 18 Olabeled phosphate. Labeled nucleotides of this type can be used to probe the details of enzyme-catalyzed reactions using the methodologies of positional isotope exchange (PIX) and molecular isotope exchange (MIX).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Manganese-Induced Substrate Promiscuity in the Reaction Catalyzed by Phosphoglutamine Cytidylyltransferase from Campylobacter jejuni

Taylor

Raushel

2019

Biochemistry

View full text Add to dashboard Cite

The leading cause of bacterial gastroenteritis, Campylobacter jejuni, is a Gram-negative pathogen that contains a unique O-methyl phosphoramidate (MeOPN) on its capsular polysaccharide. Previously, MeOPN has been linked to the evasion of host immune responses and serum resistance. Despite the involvement of MeOPN in pathogenicity, the complete biosynthesis of this modification is unknown; however, the first four enzymatic steps have been elucidated. The second enzyme in this pathway, Cj1416, is a CTP:phosphoglutamine cytididylyltransferase that catalyzes the displacement of pyrophosphate from MgCTP by L-glutamine phosphate to form CDP-Lglutamine. Initially, Cj1416 was predicted to use phosphoramidate to form cytidine diphosphoramidate but no activity was detected with MgATP as a substrate. However, in the presence of MnCTP, Cj1416 can directly catalyze the formation of cytidine diphosphoramidate from phosphoramidate and MnCTP. Here we characterize the manganese-induced promiscuity of Cj1416. In the presence of Mn 2+ , Cj1416 catalyzes the formation of 12 different reaction products using L-glutamine phosphate, phosphoramidate, methyl phosphate, methyl phosphonate, phosphate, arsenate, ethanolamine phosphate, glycerol-1-phosphate, glycerol-2-phosphate, serinol phosphate, L-serine phosphate or 3-phospho-D-glycerate as the nucleophile to displace pyrophosphate from CTP.

show abstract

Section: Resultssupporting

confidence: 86%

Section: ■ Results and Discussionmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Manganese-Induced Substrate Promiscuity in the Reaction Catalyzed by Phosphoglutamine Cytidylyltransferase from Campylobacter jejuni

Taylor

Raushel

2019

Biochemistry

View full text Add to dashboard Cite

show abstract

“…Moreover, our protocol prevents the carbonation of ribonucleotides, which is usually observed with the use of CDI . Our procedure, if applied to NDPs and NTPs, gives yields in the same order of magnitude as those obtained by anhydrous procedures,, , whereas better yields are reported in the literature for CDP‐choline and UDPCH 2 P . Last, but not least, ≈30–60 % of NMP introduced in the reaction ends up in the final product.…”

Section: Resultsmentioning

confidence: 54%

“…This approach is based on the nucleophilic substitution of nucleoside 5′‐monophosphates (NMPs) activated as phosphoramidate monoesters by suitable inorganic phosphate salts. The products obtained are nucleoside 5′‐diphosphates (NDPs), nucleoside 5′‐triphosphates (NTPs), dinucleoside 5′,5′‐polyphosphates (DNPs), and conjugates such as NDP‐sugars and cytidine 5′‐diphosphate choline (CDP‐choline) . Nucleoside phosphorimidazolide intermediates can be obtained by the reaction of NMPs with 1,1′‐carbonyldiimidazole (CDI).…”

Section: Introductionmentioning

confidence: 99%

One‐Pot Synthesis of Nucleotides and Conjugates in Aqueous Medium

2016

Self Cite

View full text Add to dashboard Cite

Herein, we describe a one‐pot synthesis, in a mixture of water/acetonitrile, of nucleoside 5′‐polyphosphates and some derivatives starting from their corresponding nucleoside 5′‐monophosphates. Phosphorimidazolide intermediates are formed in the presence of imidazole and 2‐chloro‐1,3‐dimethylimidazolinium hexafluorophosphate. Under these mild conditions, nucleoside 5′‐di‐ and 5′‐triphosphates, dinucleoside 5′,5′‐polyphosphates, as well as some nucleotide analogues modified either on the nucleoside or on the phosphate moieties are obtained in moderate to high yields.

show abstract

A cocktail of protein engineering strategies: Breaking the enzyme bottleneck one by one for high UTP production in vitro

Sun

Lou

et al. 2022

Biotech & Bioengineering

View full text Add to dashboard Cite

The pyrimidine metabolic pathway is tightly regulated in microorganisms, allowing limited success in metabolic engineering for the production of pathway‐related substances. Here, we constructed a four‐enzyme coupled system for the in vitro production of uridine triphosphate (UTP). The enzymes used include nucleoside kinase, uridylate kinase, nucleoside diphosphate kinase, and polyphosphate kinase for energy regeneration. All these enzymes are derived from extremophiles. To increase the total and unit time yield of the product, three enzymes other than polyphosphate kinase were modified separately by multiple protein engineering strategies. A nucleoside kinase variant with increased specific activity from 2.7 to 36.5 U/mg, a uridylate kinase variant (specific activity of 37.1 U/mg) with a 5.2‐fold increase in thermostability, and a nucleoside diphosphate kinase variant with a 2‐fold increase in a specific activity to over 900 U/mg were obtained, respectively. The reaction conditions of the coupled system were further optimized, and a two‐stage method was taken to avoid the problem of enzymatic pH adaptation mismatch. Under optimal conditions, this system can produce more than 65 mM UTP (31.5 g/L) in 3.0 h. The substrate conversion rate exceeded 98% and the maximum UTP productivity reached 40 mM/h.

show abstract

CDP-Ethanolamine and CDP-Choline: one-pot synthesis and 31P NMR study

Cited by 8 publications

References 37 publications

Manganese-Induced Substrate Promiscuity in the Reaction Catalyzed by Phosphoglutamine Cytidylyltransferase from Campylobacter jejuni

Manganese-Induced Substrate Promiscuity in the Reaction Catalyzed by Phosphoglutamine Cytidylyltransferase from Campylobacter jejuni

One‐Pot Synthesis of Nucleotides and Conjugates in Aqueous Medium

A cocktail of protein engineering strategies: Breaking the enzyme bottleneck one by one for high UTP production in vitro

Contact Info

Product

Resources

About