Overview

As a critical category of glycosyltransferases, mannosyltransferases (MTases) facilitate the transfer of mannose residues, serving as a cornerstone in both N-linked and O-linked glycosylation pathways. Exacting regulation of mannosylation is essential across diverse biological processes—ranging from protein folding and quality surveillance in the endoplasmic reticulum to cell-cell communication and immune system regulation. Within the biopharmaceutical sector, differences in glycosylation patterns exert a substantial influence on a therapeutic protein's effectiveness, duration of action, and potential to trigger immune responses. At CD BioGlyco, our dedicated MTase engineering service enables precise modification of these enzymes to attain target glycosylation characteristics, thus opening up novel avenues for drug discovery, biomanufacturing, and basic scientific investigation.

Key Technologies

We combine rational design with advanced genetic and molecular biology tools to precisely edit and optimize MTase function. Our core technologies include gene synthesis and directed evolution, which allow us to create extensive libraries of enzyme variants with enhanced properties. We utilize a powerful and versatile gene-editing technology, to perform targeted knockouts, knock-ins, or point mutations on specific genes encoding MTases in host cell lines.

MTase Engineering & Gene Editing: A Catalyst for Glycobiology Innovation

CD BioGlyco has rich experience in Gene Editing. We have a variety of biological models such as mice, cells, fungi, etc., and are equipped with high-standard molecular experimental platforms. We provide a full workflow from sample acquisition to gene modification detection.

MTase is commonly found in eukaryotes. Its main function is to transfer mannose from GDP-mannose (GDP-man) to long-alcohol monophosphate to form long-alcohol phosphate mannose. MTase plays an important role in organisms. It affects protein glycosylation, plant disease immunity, seedling growth, embryo development, and cancer cell migration. We offer the following types of MT gene modification services.

Fig.1 Process of MT gene modification. (CD BioGlyco)

• ALG mannosyltransferase (ALG MTase) gene modification service
  Most Alg MTases are endoplasmic reticulum membrane proteins with more functions. For example, Alg1 is mainly involved in the initial stages of protein glycosylation modification. Studies have shown that high expression of ALG1 has an inhibitory effect on the migration of hepatocellular carcinoma cells. Alg2 catalyzes the addition of both α1,3- and α1,6-linked man residues to generate the core pentasaccharide Man3GlcNAc2. We influence processes such as protein glycosylation modification and lipid synthesis by upregulating ALG expression.
• Protein O-mannosyltransferase (POMT) gene modification service
  POMT plays an important role in eukaryotic protein modification. PMT is a very large and complex family. For example, at least seven PMTs have been identified in the yeast endoplasmic reticulum. We modify the POMT gene by constructing a POMT knockout mutant.
• GDP-mannosyltransferase (GMT) gene modification service
  GMT adds mannose to glucose α(1->3) during the biosynthesis of the water-soluble branched-chain polysaccharide acetonide in the genus Acetobacter. We examine changes in sugar synthesis in GMT mutants by knockout or knockdown GMT.
• MTs heterologous expression service
  The expression of recombinant proteins using heterologous hosts has many advantages. According to the customer's requirements, we will develop a detailed experimental program. We Knock the Exogenous MT Gene into the host. We will analyze the enzyme activity, in vivo function, and changes in the host phenotype of the exogenous MT gene. Target proteins with functional properties are produced by heterologous expression of the host.

Workflow

Published Data

Paper Title: Knockout of an endogenous mannosyltransferase increases the homogeneity of glycoproteins produced in Pichia pastoris

Technology: Gene Knockout

Journal: Scientific Reports

IF: 3.998

Published: 2013

Results: The OCH1 gene encodes an α-1,6-mannosyltransferase. After the Knockout of the OCH1 gene from Pichia pastoris, it resulted in the production of recombinant proteins with a shorter glycan structure in the new P. pastoris compared to the wild type. It has an altered phenotype and growth pattern with a clumped appearance and multinodular cells.

Fig.2 Phenotypic variation in the novel P. pastoris. (Krainer, et al., 2013)

Applications

• Abnormalities in glycosyltransferases (GTs) may lead to disease development. Genetic modification of MTase is used to study the mechanisms of disease development.
• GTs are the basis and prerequisite for glycosylation modifications. Genetic modification of MTase is used to study the function of glycosylation modifications in organisms.
• The function of MTase genes in some plants is not verified. The use of gene editing tools such as overexpression and knockdown verify the function of MTase genes in this species.

Advantages

• Our experimental team is experienced in providing customized advice for various gene editing functions.
• We perform precise editing of the genome, high-efficiency knockdown, and multi-step validation to ensure reliable results.
• We monitor the progress of the project in real time and make timely adjustments to ensure that the project is completed on time.

Frequently Asked Questions

CD BioGlyco specializes in providing one-stop gene modification services for life science researchers. Besides MTase, we also provide N-Acetylglucosaminyltransferase Engineering Service and N-Acetylgalactosaminyltransferase Engineering Service. Please feel free to contact us and we will fulfill all your research needs.

Associated Services