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Glucosyltransferase Engineering Service

Glucosyltransferase Engineering Service

Diversification of the Glucosyltransferase (GT) Gene

GT catalyzes the sequential transfer of glucose from specific activated donors to specific acceptor molecules for the formation of oligosaccharides, polysaccharides, and other biomolecules. GT is a catalyst for glycan modification, complex glycans, etc. The synthesis of disaccharides, oligosaccharides, and polysaccharides involves several different GTs. Depending on the stereochemistry of the substrates and products of the reaction, GTs can be categorized as either convertases or retention enzymes. GTs are a highly diverse group of enzymes. GTs of bacterial, plant, animal, and viral origin have been classified into 116 families based on amino acid sequence similarity and catalytic mechanism.

CD BioGlyco utilizes cutting-edge gene editing technology to engineer GTs, providing a powerful approach to alter substrate specificity, improve catalytic efficiency, and develop novel glycan structures with customized properties. This advanced technology is crucial for a variety of applications, from producing complex carbohydrates for therapeutics to optimizing biofuel production.

Key Technologies

Our core expertise lies in the precise manipulation of genetic material to alter enzyme characteristics. This includes leveraging powerful gene editing systems for highly specific gene knock-in or knock-out, which is used to introduce or remove GT genes in host organisms. Additionally, we employ site-directed mutagenesis to introduce single or multiple amino acid substitutions, precisely tuning the enzyme's active site or allosteric regulation. High-throughput screening services and advanced protein expression systems in various hosts, including E. coli, yeast, and mammalian cells, are utilized to express and purify the engineered enzymes at high yields.

The Future of Glycoscience: Glucosyltransferase Engineering and Gene Editing Service

CD BioGlyco specializes in the development and application of technologies for gene editing. We are focusing on current hot gene editing topics and expanding our GT engineering service. We aim to provide better Gene Editing Services to our clients.

With the sequencing of biological genomes and continuous research in functional genomics, more and more GTs have been identified. We target the synthesis of sugars with specific functions and properties by altering GT expression to affect sugar synthesis through gene editing techniques. We edit a wide range of GT-related genes in animals, plants, and cancer. The editable GT genes include but are not limited to:

Types of editable GT genes.Fig.1 Types of editable GT genes. (CD BioGlyco)

  • Mannosyl (alpha-1,6-)-glycoprotein beta-1,6-N-acetyl-glucosaminyltransferase (MGAT5) gene modification service
    MGAT5 is associated with the synthesis of cell surface and secreted glycoprotein β1,6GlcNAc-branched N-linked glycans. MGAT5 has emerged as a promising target for cancer therapy and is involved in the stimulation of oncoproteins in cancer cells to promote carcinogenesis. We construct relevant gene expression vectors to regulate the expression of the MGAT5 gene.
  • Beta-1,3-galactosyl-O-glycosyl-glycoprotein beta-1,6-N-acetylglucosaminyltransferase (GCNT3) gene modification service
    GCNT3 has a variety of activities. For example, GCNT3 plays a key role in mucosal function and tumor migration and invasion. Researchers have found that silencing and functional inhibition of GCNT3 will greatly inhibit the migration and invasion of melanoma cells. This provides potential possibilities for the treatment of melanoma at the molecular level. We provide corresponding vector construction services. We develop and complete gene editing programs according to client needs.
  • Glycogen alpha-4-glucosyltransferase gene modification service
    GTs play an important role in glycogen synthesis and include two types. We affect the expression of glycogen alpha-4-glucosyltransferase in animals by Overexpression and Knockdown.
  • Mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (Mgat1) gene modification service
    Mgat1 is essential for the conversion of high mannose to complex and heterogeneous N-glycan synthesis, initiating the formation of complex N-linked carbohydrates. We use an emerging editing technology to affect the synthesis of specific N-glycans through the Mgat1 knockdown.
  • Other GT gene modification services
    UDP-glycosyltransferases (UGTs) catalyze the transfer of sugar groups from activated nucleotide sugar donors to acceptors to form glycosidic bonds. Glycosylation modification of natural products in plants is mainly achieved by UGTs. We also provide UGTs gene editing services.

Workflow

Workflow for GT engineering services. (CD BioGlyco)

Publication Data

Journal: Horticulture research

IF: 8.5

Published: 2019

Results: This study applied genome editing in pomegranate hairy roots to target two GTs, PgUGT84A23 and PgUGT84A24, involved in hydrolyzable tannin (HT) biosynthesis. Dual knockouts (ugt84a23 ugt84a24) showed reduced punicalagins (major HTs) and accumulated gallic acid 3-O- and 4-O-glucosides, absent in controls or single knockouts. Transcriptome and real-time quantitative polymerase chain reaction (qPCR) analyses identified 11 upregulated UGTs, with PgUGT72BD1 characterized as a regiospecific enzyme producing gallic acid 4-O-glucoside. Phylogenetically grouped in UDP-dependent glycosyltransferases (UGTs) group E, PgUGT72BD1 exhibits high substrate affinity but low catalytic efficiency compared to the targeted UGTs and is root-specific. This work establishes genome editing as a tool for pomegranate functional genomics and germplasm improvement via phytochemical modification.

Fig.2 Gene editing of GTs.Fig.2 Gene editing of PgUGT84A23 and PgUGT84A24. (Chang, et al., 2019)

Applications

  • GT is an important regulator of secondary metabolism in plants and is involved in the response to adversity stresses. The technologies for GT gene editing are used to optimize plant tolerance.
  • Technologies for GT gene editing have an important role in protein structure analysis, GT catalytic mechanism, catalytic activity modification, and other studies.
  • Technologies for GT gene editing achieve heterologous expression of recombinant proteins.

Advantages

  • We have a wide range of GT genes that are edited, diverse services, and efficient technologies for gene editing. We aim to provide our clients with high-quality gene editing services.
  • Technologies for GT gene editing help to study its effect on cancer cell migration and invasion, thus providing potential therapeutic targets for cancer treatment.
  • Technologies for GT gene editing help to effectively improve the water solubility, pharmacological activity, and bioavailability of plant natural products, which is crucial for drug development of plant natural products.

Frequently Asked Questions

CD BioGlyco relies on a professional research team and advanced technologies to provide one-stop gene editing services according to your requirements. In addition to GT, we also provide Galactosyltransferase Engineering Service and Mannosyltransferase Engineering Service. Please feel free to contact us for more information.

Associated Services

Custom Carbohydrate Synthesis

We synthesize a wide range of complex carbohydrates to be used as substrates or standards.

(AI-CD BioGlyco)

Glycoprotein Quantification

We provide detailed structural analysis to understand the glycosylation patterns.

(AI-CD BioGlyco)

Cell Line Glycoengineering

We develop stable cell lines that are optimized for the production of specific glycoconjugates.

(AI-CD BioGlyco)

Reference

  1. Chang, L.; et al. Effective genome editing and identification of a regiospecific gallic acid 4-O-glycosyltransferase in pomegranate (Punica granatum L.). Horticulture research. 2019, 6. (Open Access)
This service is for Research Use Only, not intended for any clinical use.
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CD BioGlyco is a world-class biotechnology company with offices in many countries. Our products and services provide a viable option to what is otherwise available.

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