Streamlined chemoenzymatic total synthesis of prioritized ganglioside cancer antigens

Yu, Hai; Santra, Abhishek; Li, Yanhong; McArthur, John B.; Ghosh, Tapas; Yang, Xiaoxiao; Wang, Peng George; Chen, Xi

doi:10.1039/c8ob01087k

Cited by 41 publications

(84 citation statements)

References 49 publications

(57 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All NMR data and spectra presented in the protocol match those reported previously (Yu et al, 2018). The presence of a single Neu5Ac moiety is evident from the chemical shifts of Neu5Ac H-3eq (2.91), H-3ax (1.76), and C-3 (40.66)…”

Section: Understanding Resultssupporting

confidence: 72%

“…As shown in Figure 2, lactosyl sphingosine (LacβSph; Yu et al, 2018) is used as the acceptor substrate for Pasteurella multocida α2-3-sialyltransferase 3 (PmST3; Thon, Li, Yu, Lau, & Chen, 2012) in a one-pot, two-enzyme sialylation system to synthesize Neu5Acα2-3LacβSph (GM3βSph), which is then used as the acceptor substrate for the α2-8-sialyltransferase activity of Campylobacter jejuni CstII (CjCstII; Cheng et al, 2008) in another one-pot, two-enzyme sialylation system to synthesize Neu5Acα2-8Neu5Acα2-3LacβSph (GD3βSph). In both systems, commercially available N-acetylneuraminic acid (Neu5Ac) is activated by Neisseria meningitidis CMPsialic acid synthetase (NmCSS; Yu, Yu, Karpel, & Chen, 2004) and then transferred by the appropriate sialyltransferase to the acceptor to produce the desired sialylated product.…”

Section: Chemoenzymatic Synthesis and Purification Of Gm3 And Gd3 Gangliosidesmentioning

confidence: 99%

See 1 more Smart Citation

Chemoenzymatic Synthesis and Facile Purification of Gangliosides

Bai

Chen

2021

Current Protocols

Self Cite

View full text Add to dashboard Cite

Gangliosides are biologically important sialic acid-containing glycolipids found commonly in human and other vertebrates. Isolation of pure gangliosides from cells or tissues is difficult, and chemical synthesis of gangliosides usually involves numerous steps with low synthetic yields. We report here a chemoenzymatic synthesis and purification protocol for two ganglioside cancer antigens, GM3 and GD3. One-pot multienzyme glycosylation reactions are used to sequentially prepare GM3 and GD3 sphingosines from chemically synthesized lactosyl sphingosine. A facile C18-cartridge purification procedure after each glycosylation reaction provides the desired pure glycosyl sphingosine product, which is readily acylated to form the target ganglioside.

show abstract

Section: Understanding Resultssupporting

confidence: 72%

Section: Chemoenzymatic Synthesis and Purification Of Gm3 And Gd3 Gangliosidesmentioning

confidence: 99%

Chemoenzymatic Synthesis and Facile Purification of Gangliosides

Bai

Chen

2021

Current Protocols

Self Cite

View full text Add to dashboard Cite

show abstract

“…It is evident that under thermodynamic control nucleotide regeneration and enzymatic glycosylation can only occur with highly exergonic sacrificial substrates (i.e., hydrolysis of pyrophosphate), as was proposed by Hirschbein et al who compared the energy of hydrolysis of the sacrificial donors as a rationale for glycosylation efficiency [203]. Besides, for the common nucleotide glycosylation donors UDP-Glc, UDP-GlcNAc [204], UDP-GlcA [205], UDP-Gal [204], UDP-GalA, UDP-Xyl, GDP-Man, GDP-Fuc [204], CMP-Neu5Ac [200,204] the (re)generation systems for the production have been employed for rare or synthetic nucleotide sugar donors, such as CMP-MAnNGc [200], CMP-Man [200], CMP-ManNac5OMe [200], CMP-Kdo [200], ADP-Hep [206], and dTDP-Rha [207].…”

Section: Application Of Glycosyl Transferases In Organic Synthesismentioning

confidence: 99%

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Mestrom

Przypis

Kowalczykiewicz

et al. 2019

IJMS

View full text Add to dashboard Cite

Enzymes are nature's catalyst of choice for the highly selective and efficient coupling of carbohydrates. Enzymatic sugar coupling is a competitive technology for industrial glycosylation reactions, since chemical synthetic routes require extensive use of laborious protection group manipulations and often lack regio-and stereoselectivity. The application of Leloir glycosyltransferases has received considerable attention in recent years and offers excellent control over the reactivity and selectivity of glycosylation reactions with unprotected carbohydrates, paving the way for previously inaccessible synthetic routes. The development of nucleotide recycling cascades has allowed for the efficient production and reuse of nucleotide sugar donors in robust one-pot multi-enzyme glycosylation cascades. In this way, large glycans and glycoconjugates with complex stereochemistry can be constructed. With recent advances, LeLoir glycosyltransferases are close to being applied industrially in multi-enzyme, programmable cascade glycosylations.hydroxynitrile lyases catalyzed the enzymatic hydrolysis of the glycoside amygdalin [3]. Moving almost two centuries forward, the largest volumetric biocatalytic industrial process is the application of glucose isomerase for the production of high fructose syrup for food and drink applications, producing fructose from glucose at 10 7 tons per year [4]. The secret of the success of enzymes in the production or treatment of carbohydrates and glycosides is their exquisite stereo-and regioselectivity. The excellent selectivity of enzymes is required due to the diversity of structural features of carbohydrates [5], comprising d-and l-epimers, ring size, anomeric configuration, linkages, branching, and oxidation state(s). Since drug targets often exhibit specificity for all of these structural features, the production process should not contain any side-products to prevent undesired side-effects [6].The challenge in the synthesis of carbohydrates is their wide variety of functionalities and stereochemistry ( Figure 1). (Poly)hydroxyaldehydes containing a terminal aldehyde are referred to as aldoses and (poly)hydroxyketones are defined as ketoses. In aqueous solutions, monosaccharides form equilibrium mixtures of linear open-chain and ring-closed 5-or 6-membered furanoses or pyranoses, respectively. For aldoses, the asymmetric ring forms at C-1. For ketoses, it closes at C-2 as an axial (α) or equatorial (β) hemiacetal or hemiketal, respectively (commonly defined as the anomeric center). A glycosidic linkage is a covalent O-, S-, N-, or C-bond connecting a monosaccharide to another residue resulting in a glycoside, while glucoside is specific for a glucose moiety. The equatorial or axial position of the glycosidic bond is referred to as α-(axial) or β-linkage (equatorial). The number of carbohydrates linked via glycosidic bonds can be subdivided into oligosaccharides with two to ten linked carbohydrates, while polysaccharides (glycans) contain more than ten glycosidic bonds. A glycan either con...

show abstract

“…These oligosaccharides have been chemically synthesized by time-consuming protocols involving more than 50 chemical steps including difficult sialylations that gave mixtures of anomers requiring tedious separation by silica gel column chromatography 49,50 . Gangliosides, such as GM1, GD1b and fucosyl GM1, have been synthesized by elegant (chemo)enzymatic approaches, but purification relied on time consuming size exclusion column or reverse phase column chromatography followed by lyophilization of product containing fractions [51][52][53] . Finally, the successful preparation of N-glycan 22 (Fig.…”

Section: Discussionmentioning

confidence: 99%