Engineered GH for Improved Glycan Polymer Synthesis

Research output: Innovation

Abstract


Invention Summary:

Oligosaccharides and polysaccharides (or glycans) are complex macromolecules in living systems that serve critical structural and functional roles. In vitro synthesis of these complex biomolecules is very challenging. Glycosytransferases are naturally available enzymes that catalyze the synthesis of glycans, but have poor expression yield, narrow substrate specificity, and use expensive nucleotide sugars.

Rutgers scientists have engineered a multifunctional family 5 glycosyl hydrolase (GH-5 called CelE) from Clostridium thermocellum into a transglycosidase to synthesize diverse glycan-based polymers (e.g., glucans, xylans, mannans). This enzyme is composed of an active site-mutant (CelE-E316G) fused with a unique carbohydrate-binding module and linker peptide at its C-terminus (called CBM3a).


The transglycosylation activity of CelE-E316G is dramatically enhanced when fused with CBM3a to produce cellotriose and cellotetraose from pNP-cellobiose, as demonstrated by the pNP release rate shown in the figure above.

Market Applications:

Production of designer oligosaccharides containing specific monosaccharides and unique glycosidic linkages using engineered enzymes.

Advantages:

  • High transglycosylation activity
  • High expression yield

Intellectual Property & Development Status:

Patent pending. Available for licensing and/or research collaboration.

Original languageEnglish (US)
StatePublished - Mar 2019

Fingerprint

Polysaccharides
Polymers
Oligosaccharides
Enzymes
Clostridium thermocellum
Intellectual Property
Mannans
Cellobiose
Xylans
Glucans
Monosaccharides
Hydrolases
Licensure
Substrate Specificity
Catalytic Domain
Nucleotides
Carbohydrates
Peptides
Research

Cite this

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title = "Engineered GH for Improved Glycan Polymer Synthesis",
abstract = "Invention Summary: Oligosaccharides and polysaccharides (or glycans) are complex macromolecules in living systems that serve critical structural and functional roles. In vitro synthesis of these complex biomolecules is very challenging. Glycosytransferases are naturally available enzymes that catalyze the synthesis of glycans, but have poor expression yield, narrow substrate specificity, and use expensive nucleotide sugars. Rutgers scientists have engineered a multifunctional family 5 glycosyl hydrolase (GH-5 called CelE) from Clostridium thermocellum into a transglycosidase to synthesize diverse glycan-based polymers (e.g., glucans, xylans, mannans). This enzyme is composed of an active site-mutant (CelE-E316G) fused with a unique carbohydrate-binding module and linker peptide at its C-terminus (called CBM3a). The transglycosylation activity of CelE-E316G is dramatically enhanced when fused with CBM3a to produce cellotriose and cellotetraose from pNP-cellobiose, as demonstrated by the pNP release rate shown in the figure above. Market Applications: Production of designer oligosaccharides containing specific monosaccharides and unique glycosidic linkages using engineered enzymes. Advantages: High transglycosylation activity High expression yield Intellectual Property & Development Status: Patent pending. Available for licensing and/or research collaboration.",
author = "Shishir Chundawat",
year = "2019",
month = "3",
language = "English (US)",
type = "Patent",

}

TY - PAT

T1 - Engineered GH for Improved Glycan Polymer Synthesis

AU - Chundawat, Shishir

PY - 2019/3

Y1 - 2019/3

N2 - Invention Summary: Oligosaccharides and polysaccharides (or glycans) are complex macromolecules in living systems that serve critical structural and functional roles. In vitro synthesis of these complex biomolecules is very challenging. Glycosytransferases are naturally available enzymes that catalyze the synthesis of glycans, but have poor expression yield, narrow substrate specificity, and use expensive nucleotide sugars. Rutgers scientists have engineered a multifunctional family 5 glycosyl hydrolase (GH-5 called CelE) from Clostridium thermocellum into a transglycosidase to synthesize diverse glycan-based polymers (e.g., glucans, xylans, mannans). This enzyme is composed of an active site-mutant (CelE-E316G) fused with a unique carbohydrate-binding module and linker peptide at its C-terminus (called CBM3a). The transglycosylation activity of CelE-E316G is dramatically enhanced when fused with CBM3a to produce cellotriose and cellotetraose from pNP-cellobiose, as demonstrated by the pNP release rate shown in the figure above. Market Applications: Production of designer oligosaccharides containing specific monosaccharides and unique glycosidic linkages using engineered enzymes. Advantages: High transglycosylation activity High expression yield Intellectual Property & Development Status: Patent pending. Available for licensing and/or research collaboration.

AB - Invention Summary: Oligosaccharides and polysaccharides (or glycans) are complex macromolecules in living systems that serve critical structural and functional roles. In vitro synthesis of these complex biomolecules is very challenging. Glycosytransferases are naturally available enzymes that catalyze the synthesis of glycans, but have poor expression yield, narrow substrate specificity, and use expensive nucleotide sugars. Rutgers scientists have engineered a multifunctional family 5 glycosyl hydrolase (GH-5 called CelE) from Clostridium thermocellum into a transglycosidase to synthesize diverse glycan-based polymers (e.g., glucans, xylans, mannans). This enzyme is composed of an active site-mutant (CelE-E316G) fused with a unique carbohydrate-binding module and linker peptide at its C-terminus (called CBM3a). The transglycosylation activity of CelE-E316G is dramatically enhanced when fused with CBM3a to produce cellotriose and cellotetraose from pNP-cellobiose, as demonstrated by the pNP release rate shown in the figure above. Market Applications: Production of designer oligosaccharides containing specific monosaccharides and unique glycosidic linkages using engineered enzymes. Advantages: High transglycosylation activity High expression yield Intellectual Property & Development Status: Patent pending. Available for licensing and/or research collaboration.

UR - http://rutgers.technologypublisher.com/tech/Engineered_GH_for_Improved_Glycan_Polymer_Synthesis

M3 - Innovation

ER -