Highly efficient chemoenzymatic synthesis and facile purification of α-Gal pentasaccharyl ceramide Galα3nLc<sub>4</sub>βCer

Santra, Abhishek; Li, Yanhong; Yu, Hai; Slack, Teri J.; Wang, Peng George; Chen, Xi

doi:10.1039/c7cc04090c

Cited by 25 publications

(35 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Solution-phase synthesis with a substrate bound to water-soluble [224] or thermo-responsive polymers [225], fluorous- [217,226,227,228,229,230,231,232,233,234,235,236], ion exchange [237], or lipid-like tags [238] has attracted much interest since it can bypass compatibility issues between enzymes and solid carriers. The main disadvantage of solution-phase assembly of oligosaccharides catalyzed by glycosyltransferases is the low catalytic efficiency and affinity for the substrates due to different steric and stereoelectronic properties induced by the substrate-bound tag.…”

Section: Reactor Engineering For (Non)-leloir Glycosyltransferasesmentioning

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

Section: Reactor Engineering For (Non)-leloir Glycosyltransferasesmentioning

confidence: 99%

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Mestrom

Przypis

Kowalczykiewicz

et al. 2019

IJMS

View full text Add to dashboard Cite

show abstract

“…Recently, we showed that α-Gal pentasaccharyl ceramide, a neutral glycosphingolipid, can be chemoenzymatically synthesized from water soluble lactosyl sphingosine (LacβSph) with simple C18-cartridge solid phase extraction (SPE) purification procedures followed by acylation. 23 Herein, we develop high-yield streamlined chemoenzymatic processes for total synthesis of synthetically challenging complex gangliosides. A 13-gram-scale synthetic procedure is established for producing LacβSph from commercially available inexpensive lactose and phytosphingosine.…”

mentioning

confidence: 99%

“…To obtain target gangliosides, LacβSph ( 5 ) 23 was prepared from lactose and phytosphingosine in a 13-gram scale to demonstrate the efficiency of the streamlined chemical synthetic procedure and to provide a sufficient amount of starting material for glycosphingolipid synthesis. The synthesis was achieved in 12 steps in an overall 40% yield (see ESI).…”

mentioning

confidence: 99%

Streamlined chemoenzymatic total synthesis of prioritized ganglioside cancer antigens

Santra

et al. 2018

Org. Biomol. Chem.

Self Cite

View full text Add to dashboard Cite

A highly efficient streamlined chemoenzymatic strategy for total synthesis of four prioritized ganglioside cancer antigens GD2, GD3, fucosyl GM1, and GM3 from commercially available lactose and phytosphingosine is demonstrated. Lactosyl sphingosine (LacβSph) was chemically synthesized (on a 13 g scale), subjected to sequential one-pot multienzyme (OPME) glycosylation reactions with facile C18-cartridge purification, followed by improved acylation conditions to form target gangliosides, including fucosyl GM1 which has never been synthesized before.

show abstract

“…The sphingosine component facilitated the facile C18cartridge-based purification processes (Figure 14). Ganglio-and neolacto-series glycosphingolipids have been successfully obtained using the strategy [67,68]. Compared to en bloc transfer of oligosaccharides from chemoenzymatically synthesized oligosaccharyl fluorides for the synthesis of glycosphingolipids and glycosphingosines containing a sphingoid base or its derivative using endoglycoceramidase mutants (endoglycoceramidase glycosynthase) [69,70], the OPME glycosylation of LacβSph strategy is more flexible in accessing a more diverse array of products and also provides an easier C18 cartridgepurification process.…”

Section: Chemoenzymatic Synthesis Of Glycolipids With Facile Purificamentioning

confidence: 99%

Strategies for chemoenzymatic synthesis of carbohydrates

McArthur

Chen

2019

Carbohydrate Research

Self Cite

View full text Add to dashboard Cite

Carbohydrates are structurally complex but functionally important biomolecules. Therefore, they have been challenging but attractive synthetic targets. While substantial progress has been made on advancing chemical glycosylation methods, incorporating enzymes into carbohydrate synthetic schemes has become increasingly practical as more carbohydrate biosynthetic and metabolic enzymes as well as their mutants with synthetic application are identified and expressed for preparative and large-scale synthesis. Chemoenzymatic strategies that integrate the flexibility of chemical derivatization with enzyme-catalyzed reactions have been extremely powerful. Briefly summarized here are our experiences on developing one-pot multienzyme (OPME) systems and representative chemoenzymatic strategies from others using glycosyltransferase-catalyzed reactions for synthesizing diverse structures of oligosaccharides, polysaccharides, and glycoconjugates. These strategies allow the synthesis of complex carbohydrates including those containing naturally occurring carbohydrate postglycosylational modifications (PGMs) and nonnatural functional groups. By combining these strategies with facile purification schemes, synthetic access to the diverse space of carbohydrate structures can be automated and will not be limited to specialists.

show abstract

Highly efficient chemoenzymatic synthesis and facile purification of α-Gal pentasaccharyl ceramide Galα3nLc₄βCer

Cited by 25 publications

References 51 publications

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Streamlined chemoenzymatic total synthesis of prioritized ganglioside cancer antigens

Strategies for chemoenzymatic synthesis of carbohydrates

Contact Info

Product

Resources

About

Highly efficient chemoenzymatic synthesis and facile purification of α-Gal pentasaccharyl ceramide Galα3nLc4βCer

Cited by 25 publications

References 51 publications

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach

Streamlined chemoenzymatic total synthesis of prioritized ganglioside cancer antigens

Strategies for chemoenzymatic synthesis of carbohydrates

Contact Info

Product

Resources

About

Highly efficient chemoenzymatic synthesis and facile purification of α-Gal pentasaccharyl ceramide Galα3nLc₄βCer