Bacteria Glycoengineering Services

Services Published Data

Native Glycosylation in Bacteria

Native glycosylation pathways have been discovered in certain bacterial species. Pgl from the bacterium Campylobacter jejuni is the first characterized N-glycosylation pathway. In this pathway, undecaprenyl phosphate lipid (Und-PP)-linked glycans are transferred by oligosaccharyltransferase (OST) known as PglB. PglB is a single-subunit integral membrane protein that shares significant sequence similarity with the catalytic subunit of the eukaryotic OST, which is referred to as STT3.

Bacterial O-linked glycosylation pathways share similarities with the N-linked pathways. Specific bacteria like Neisseria meningitidis and Neisseria gonorrhoeae utilize O-glycosylation to modify their pilins by two distinct OSTs, PglL and PglO, respectively. The key distinction from eukaryote glycosylation lies in the mode of glycan transfer, where the OST transfers the entire glycan in a single unit from the Und-PP carrier directly onto target proteins.

Bacteria Glycoengineering Services at Creative Biolabs

Since the initial discovery of N-glycosylation in bacteria, glycoengineering in bacterial hosts for the production of human therapeutics has undergone significant evolution. To obtain glycoprotein with tailored glycan structures, Creative Biolabs has developed various bacteria glycoengineering services, especially in Escherichia coli, based on advanced techniques for Genetic Glycoengineering. Our strategies primarily include the introduction of heterologous glycosylation pathways into bacteria, such as genes for glycan synthesis, transporters, glycosidases, and glycosyltransferases, aiming to establish novel pathways for bottom-up synthesis of both N- and O-linked glycosylation.

Our Specific Services in Bacteria Glycoengineering

N-Linked Glycoengineering in Bacteria

We generate N-linked glycoproteins by transferring the Pgl glycosylation pathway into bacteria like E. coli. And multiple enzymes have been introduced to obtain fully humanized glycoproteins.

O-Linked Glycoengineering in Bacteria

Based on native PglL/O paradigm, we engineer human-like O-glycosylation pathways into bacteria, enabling precise and efficient site-specific O-glycosylation of human glycoproteins.

Glycoengineered Bacteria: Applications and Benefits

Glycoengineered bacterial species have gained widespread use for the production of vaccines and therapeutic glycoproteins due to cost reduction, streamlined bioprocessing, and enhanced customization. Additionally, bacteria can achieve higher product titers and reduced risk of viral contamination. In conclusion, these advantages render bacterial systems an appealing platform for glycoprotein expression after glycoengineering.

Highlights of Our Services

Optional strategies for bacteria glycoengineering
Cutting-edge technologies for genetic modulation
Efficient introduction and establishment of novel glycosylation pathways
High-yield production of homogenous glycoproteins
Customized solutions for specific needs

Published data

Heparin is an important anticoagulant drug widely applied in the medical field for the prevention and treatment of thrombosis. The activity and stability of heparin sulfotransferase are crucial to the synthesis process of heparin. In this study, the authors used metabolically engineered Escherichia coli K5 as a matrix, added a monosaccharide called N-trifluoroacetylglucosamine to it, and used an efficient protein platform to produce a glycoengineered N-sulfotransferase. Moreover, they measured the stability and activity of the produced enzyme. This study provided new ideas for the in vitro synthesis strategy of heparin.

Fig.1 Heparin synthesis process using in vitro/in vivo combined strategy. Fig.1 In vitro/in vivo synthesis strategy of heparin.¹

Glycoengineering in bacteria is an active area of research and development. Creative Biolabs has pioneered the services in bacteria glycoengineering designed for the production of glycoproteins with customized glycan structures. If you have any requirements in bacteria glycoengineering, please feel free to contact us for more detailed information.

Reference

Deng, Jian-Qun, et al. "Biosynthetic production of anticoagulant heparin polysaccharides through metabolic and sulfotransferases engineering strategies." Nature Communications 15.1 (2024): 3755. Distributed under Open Access license CC BY 4.0, without modification.

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