Alpha2-6-Linkage-Specific Sialidase Mutants
Abstract
Researchers at the University of California, Davis have developed alpha2-6-linkage-specific sialidase mutants with good enzyme activity.
Full Description
Sialidases are crucial tools for determining presence and subsequent structural and functional characteristics of sialic-acid containing glycans. Although they can be used broadly and are mild enough for functional sialic acid studies or modification in biological samples, current sialidases have either a specificity towards alpha2-3-linked sialic acids or broad promiscuity towards sialic acids with alpha2-3-, alpha2-6- and alpha2-8-linkages. This limits the overall ability to study alpha2-6-linkage specific sialidases and sialic acid-containing biomolecules. There is a need for an alpha2-6-linkage-specific sialidases with good enzymatic activity.
Researchers at the University of California, Davis have developed alpha2-6-linkage-specific sialidase mutants with efficient ability to cleave various forms of sialic acid. The inventors have used structure-guided saturation mutagenesis to engineer photobacterium for improved alpha2-6-linkage sialidase specific activity. The mutant sialidase was successfully tested to show 101-fold improvement alpha2-6 sialidase activity over non-mutant sialidases. Additionally, the method used is applicable to different sialic-acid specific linkages and the sialidase mutants open up their use, in addition to glycan analysis, in therapeutic glycoproteins and vaccines.
Applications
- Characterization of sialic acids on sensitive biological samples
- Structural and functional characterization of alpha2-6-linkage sialic acid-containing carbohydrates and glycoconjugates
- Enhance the immunogenicity of conjugated vaccines
- Improve serum half-life of circulating therapeutic glycoproteins
Features/Benefits
- Photobacteria-derived sialidases
- Generated using sequential saturation mutagenesis and screened using high-throughput blue-white colony assay
- Method may be applied to different sialic acid-specific linkages to develop sialidase mutants
Patent Status
| Country | Type | Number | Dated | Case |
| United States Of America | Published Application | 2020-032545 | 10/15/2020 | 2018-318 |
Contact
- Pooja N. Bhayani
- pnbhayani@ucdavis.edu
- tel: View Phone Number.
Inventors
- Chen, Xi
- Fisher, Andrew J
- McArthur, John
Other Information
Keywords
sialidase, neosialidase, neuraminidase, engineered glucosyltransferase, glycomics, directed evolution
Additional Technologies by these Inventors
- Purification of Glycosphingosines and Glycosphingolipids
- A Photobacterium Sp. Alpha2-6-Sialytransferase 9Psp2.6St) A366g Mutant With Increased Expression Level And Improved Activity In Sialylating Tn Antigen
- Synthesis of Capsular Polysaccharides
- Legionaminic Acid Glycosyltransferases for Chemoenzymatic Synthesis of Glycans and Glycoconjugates
- Using Escherichia coli to Produce Human Milk Oligosaccharide Lactodifucotetraose
- 4-N-Derivatized Sialic Acids and Related Sialosides
- Substrate And Process Engineering For Biocatalytic Synthesis And Facile Purification Of Human Milk Oligosaccharides (HMOs)
- O-Acetyl Glycosphingosines and Gangliosides, as well as Their N-Acetyl Analogs
- Site Directed DNA Editing with Adenosine Deaminases that Act on RNA (ADAR) Enzymes
- Stable N-acetylated analogs of Sialic Acids and Sialosides
- Alpha1–2-Fucosyltransferase for Enzymatic Synthesis of Alpha1–2-linked Fucosylated Glycans
- Engineering Pasteurella Multocida Heparosan Synthase 2 (Pmhs2) For Efficient Synthesis Of Heparosan Heparin And Heparan Sulfate Oligosaccharides
- One-Pot Multienzyme Synthesis of Sialidase Reagents, Probes and Inhibitors
- Novel Methods For Chemical Synthesis Of Lactosyl Sphingosines, Glucosylsphingosines, Galactosylsphingosines, And 3-O-Sulfogalactosylsphingosines
