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.

We Are the Best Choice

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:

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 Zinc-Finger Nuclease (ZFN)-mediated Knockdown of Mgat1.
• 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.

Published Data

Paper Title: A glycosyltransferase gene responsible for pullulan biosynthesis in Aureobasidium melanogenum P16

Technology: Gene Knockout, Gene Overexpression

Journal: International journal of biological macromolecules

IF: 5.162

Published: 2017

Results: The GT gene (UGT1), which is involved in the biosynthesis of pullulan, was cloned from Aureobasdium melanogenum P16. After the knockdown of UGT1, the results showed that the knockout mutant produced much lower levels of pullulan and GT activity than its wild-type strain. At the same time, the transcript levels of other enzyme genes related to pullulan synthesis were increased. Overexpression of UGT1 revealed enhanced GT activity and did not affect the growth of A. melanogenum itself. After the overexpression strain was cultured in the fermenter, the production of pullulan was higher up to 80.0 g/L, and the molecular weight was also increased.

Fig.2 Changes in enzyme activity, pullulan, and growth of yeast itself after UGT1 knockdown (A) and overexpression (B). (Chen, et al., 2017)

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.

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.

References:

1. Chen, X.; et al. A glycosyltransferase gene responsible for pullulan biosynthesis in Aureobasidium melanogenum P16. International Journal of Biological Macromolecules. 2017, 95: 539-549.
2. Sumardika, I.W.; et al. Beta-1,3-galactosyl-O-glycosyl-glycoprotein-1,6-N-acetylglucosaminyltransferase 3 increases MCAM stability, which enhances S100A8/A9-mediated cancer motility. Oncology Research. 2018, 3(26): 431-444.