133-89-1Relevant articles and documents
Production of galactinol from sucrose by plant enzymes.
Wakiuchi, Nariaki,Shiomi, Ryohei,Tamaki, Hajime
, p. 1465 - 1471 (2003)
Galactinol, 1-O-(alpha-D-galactopyranosyl)-myo-inositol, was produced from sucrose as a starting material. UDP-Glc was prepared with sucrose and UDP using sucrose synthase partially purified from sweet potato roots. Then, the UDP-Glc was converted to UDP-Gal using yeast UDP-Gal 4-epimerase from a commercial source. Finally, galactinol was produced from the UDP-Gal and myo-inositol using galactinol synthase partially purified from cucumber leaves. The product was identified as galactinol by the retention times of HPLC, alpha-galactosidase digestion, and NMR spectrometry.
Transcriptome-wide identification of sucrose synthase genes in Ornithogalum caudatum
Li, Li-Na,Kong, Jian-Qiang
, p. 18778 - 18792 (2016)
OCAP-2-1, OCAP-2-2, OCAP-3-1 and OCAP-3-3, four glucose-containing polysaccharides from Ornithogalum caudatum, exhibit antitumor activity, suggesting their potential application as natural antitumor drugs. Although the incorporation of glucose into these polysaccharides from UDP-d-glucose is reasonably well understood, the cDNA isolation and functional characterization of genes responsible for UDP-d-glucose biosynthesis from O. caudatum has not been identified. Here, we present a full characterization of the sucrose synthase family, a Leloir glycosyltransferase responsible for UDP-d-glucose biosynthesis from O. caudatum. Specifically, a transcriptome-wide search for Sus genes in O. caudatum was first performed in the present study. A total of 5 unigenes sharing high sequence identity with Sus were retrieved from transcriptome sequencing. Three full-length Sus-like candidates derived from this unigene assembly were then obtained and isolated by reverse transcription polymerase chain reaction (RT-PCR) from O. caudatum. Additional analysis showed two conserved domains (sucrose synthase and glycosyl transferase domains) were present in this family. Phylogenetic analysis indicated that the OcSus1 and OcSus2 could be clustered together into a monocots specific clade, while OcSus3 could be classified into M & D1 category with members from the monocots and dicots species, displaying an evolutionary consistency with other plant species. These candidate isoenzymes were screened by functional expression in E. coli individually as standalone enzymes. All three cDNAs were identified to be bona fide genes and encoded sucrose synthase with varied kinetic properties. To further explore the possible role of these Sus proteins in polysaccharide biosynthesis, transcript profiles of the three genes were subsequently examined by real-time quantitative PCR in various tissues. OcSus1 and OcSus2 were therefore assumed to be responsible for the biosynthesis of the four glucose-containing polysaccharides due to their expression profiles in O. caudatum. Taken together, these data provide further comprehensive knowledge for polysaccharide biosynthesis in O. caudatum and broaden the potential application of Sus in metabolic engineering or synthetic biology as a potential gene part.
Efficient biosynthesis of uridine diphosphate glucose from maltodextrin by multiple enzymes immobilized on magnetic nanoparticles
Dong, Qing,Ouyang, Li-Ming,Yu, Hui-Lei,Xu, Jian-He
, p. 1622 - 1626 (2010)
Uridine diphosphate glucose (UDP-Glc) serves as a glucosyl donor in many enzymatic glycosylation processes. This paper describes a multiple enzyme, one-pot, biocatalytic system for the synthesis of UDP-Glc from low cost raw materials: maltodextrin and uridine triphosphate. Three enzymes needed for the synthesis of UDP-Glc (maltodextrin phosphorylase, glucose-1-phosphate thymidylytransferase, and pyrophosphatase) were expressed in Escherichia coli and then immobilized individually on amino-functionalized magnetic nanoparticles. The conditions for biocatalysis were optimized and the immobilized multiple-enzyme biocatalyst could be easily recovered and reused up to five times in repeated syntheses of UDP-Glc. After a simple purification, approximately 630 mg of crystallized UDP-Glc was obtained from 1 l of reaction mixture, for a moderate yield of around 50% (UTP conversion) at very low cost.
Enzyme Module Systems for the Synthesis of Uridine 5′-Diphospho-α- D -glucuronic Acid and Non-Sulfated Human Natural Killer Cell-1 (HNK-1) Epitope
Engels, Leonie,Henze, Manja,Hummel, Werner,Elling, Lothar
, p. 1751 - 1762 (2015)
Tailor-made strategies for the stereo- and regioselective multi-step enzymatic synthesis of glycoconjugates require well characterized glycosyltransferases and carbohydrate modifying enzymes. We here report on a novel enzyme cascade for the synthesis of uridine 5′-diphospho-α-D-glucuronic acid (UDP-GlcA) and the non-sulfated human natural killer cell-1 (HNK-1) epitope including in situ regeneration of UDP-GlcA and the cofactor nicotinamide adenine dinucleotide NAD+ by the combination of four enzymes in one-pot. In the first enzyme module sucrose synthase 1 (SuSy1) is used to produce uridine 5′-diphospho-α-D-glucose (UDP-Glc) from sucrose and uridine 5′-diphosphate (UDP). The combination with UDP-Glc dehydrogenase in the second enzyme module leads to the synthesis of UDP-GlcA with concomitant in situ regeneration of the cofactor NAD+ by nicotinamide adenine dinucleotide hydride (NADH)-oxidase. In the third enzyme module the mammalian glucuronyltransferase GlcAT-P catalyzes the synthesis of the non-sulfated HNK-1 epitope by regioselective transfer of GlcA onto N-acetyllactosamine type 2 (LacNAc type 2). We present a comprehensive study on substrate kinetics, substrate specificities, variation and relation of enzyme activities as well as cross inhibition of intermediate products. With optimized reaction conditions we obtain superior product yields with streamlined synthesis costs for the expensive nucleotide sugar UDP-GlcA and cofactor NAD+.
A chemoenzymatic route to synthesize unnatural sugar nucleotides using a novel N-acetylglucosamine-1-phosphate pyrophosphorylase from Camphylobacter jejuni NCTC 11168
Fang, Junqiang,Xue, Mengyang,Gu, Guofeng,Liu, Xian-Wei,Wang, Peng George
, p. 4303 - 4307 (2013)
A novel N-acetylglucosamine-1-phosphate pyrophosphorylase was identified from Campylobacter jejuni NCTC 11168. An unprecedented degree of substrate promiscuity has been revealed by systematic studies on its substrate specificities towards sugar-1-P and NTP. The yields of the synthetic reaction of seven kinds of sugar nucleotides catalyzed by the enzyme were up to 60%. In addition, the yields of the other nine were around 20%. With this enzyme, three novel sugar nucleotide analogs were synthesized on a preparative scale and well characterized.
Systematic study on the broad nucleotide triphosphate specificity of the pyrophosphorylase domain of the N-acetylglucosamine-1-phosphate uridyltransferase from Escherichia coli K12
Fang, Junqiang,Guan, Wanyi,Cai, Li,Gu, Guofeng,Liu, Xianwei,Wang, Peng George
, p. 6429 - 6432 (2009)
N-Acetylglucosamine-1-phosphate uridyltransferase (GlmU) from Escherichia coli K12 is a bifunctional enzyme that catalyzes both the acetyltransfer and uridyltransfer reactions in the prokaryotic UDP-GlcNAc biosynthetic pathway. In this study, we report th
Identification and characterization of a strict and a promiscuous N-acetylglucosamine-1-P uridylyltransferase in Arabidopsis
Yang, Ting,Echols, Merritt,Martin, Andy,Bar-Peled, Maor
, p. 275 - 284 (2010)
UDP-GlcNAc is an essential precursor for glycoprotein and glycolipid synthesis. In the present study, a functional nucleotidyltransferase gene from Arabidopsis encoding a 58.3 kDa GlcNAc1pUT-1 (N-acetylglucosamine-1-phosphate uridylyltransferase) was identified. In the forward reaction the enzyme catalyses the formation of UDP-N-acetylglucosamine and PPi from the respective monosaccharide 1-phosphate and UTP. The enzyme can utilize the 4-epimer UDP-GalNAc as a substrate as well. The enzyme requires divalent ions (Mg2+ or Mn2+) for activity and is highly active between pH 6.5 and 8.0, and at 30-37°C. The apparent Km values for the forward reaction were 337 μM (GlcNAc-1-P) and 295 μM (UTP) respectively. Another GlcNAc1pUT-2, which shares 86%amino acid sequence identity with GlcNAc1pUT-1, was found to convert, in addition to GlcNAc-1-P and GalNAc-1-P, Glc-1-P into corresponding UDP-sugars, suggesting that subtle changes in the UT family cause different substrate specificities. A three-dimensional protein structure model using the human AGX1 as template showed a conserved catalytic fold and helped identify key conserved motifs, despite the high sequence divergence. The identification of these strict and promiscuous gene products open a window to indentify new roles of amino sugar metabolism in plants and specifically their role as signalling molecules. The ability of GlcNAc1pUT-2 to utilize three different substrates may provide further understanding as to why biological systems have plasticity. The Authors.
Combined enzymatic synthesis of nucleotide (deoxy) sugars from sucrose and nucleoside monophosphates
Zervosen, Astrid,Stein, Andreas,Adrian, Holger,Elling, Lothar
, p. 2395 - 2404 (1996)
The synthesis of NDP-glucose 3a-d (N = A, C, U, dU) with sucrose synthase B was combined with the enzymatic synthesis of nucleoside diphosphates 2a-d from their corresponding nucleoside monophosphates 1a-d by different kinases A. Further combination with
Large scale enzymatic synthesis of oligosaccharides and a novel purification process
Zhou, Guangyan,Liu, Xianwei,Su, Doris,Li, Lei,Xiao, Min,Wang, Peng G.
, p. 311 - 314 (2011)
Herein we report the practical chemo enzymatic synthesis of trisaccharide and derivatives of iGb3 and Gb3, and a novel purification process using immobilized yeast to remove the monosaccharide from the reaction mixture. High purity oligosaccharide compoun
Catalytic reversibility of Pyrococcus horikoshii trehalose synthase: Efficient synthesis of several nucleoside diphosphate glucoses with enzyme recycling
Ryu, Soo-In,Kim, Jeong-Eun,Kim, Eun-Joo,Chung, Seung-Kyung,Lee, Soo-Bok
, p. 128 - 134 (2011)
The trehalose synthase (TreT) from Pyrococcus horikoshii represented reversible catalysis in alternative synthesis of trehalose and nucleoside 5′-diphosphate-glucose (NDP-Glc), depending on the substrates involved. TreT from P. horikoshii had differential preferences on NDP-Glc as a donor for trehalose synthesis, in which guanosine 5′-diphosphate (GDP)-Glc was the most favored in terms of reaction specificity, kcat/Km. Uridine 5′-diphosphate (UDP)- and adenosine 5′-diphosphate (ADP)-Glcs were employed with less preferences. This enzyme reversely cleaved trehalose to transfer the glucosyl moiety to various NDPs, efficiently producing NDP-Glcs. Although ADP-Glc was the least favorable donor, the counterpart, ADP, was the most favorable acceptor for the reverse synthesis of NDP-Glc in k cat/Km. GDP and UDP were less preferred, compared to ADP. In a batch reaction of 12 h, the molar yield of NDP-Glc per NDP used was decreased approximately in the order of ADP-Glc > GDP-Glc > cytidine 5′-diphosphate (CDP)-Glc or UDP-Glc. The overall productivity of the enzyme was largely improved in a gram scale for NDP-Glcs using repetitive batch reactions with enzyme recycling. Thus, it is suggested that TreT from P. horikoshii may be useful for the regeneration of NDP-Glc from NDP, especially for ADP-Glc from ADP, with trehalose as a glucose resource.
