Production of HMO by Biocatalytic Methods

Production of HMO by Biocatalytic Methods

CD BioGlyco provides a high level of technical support in fermentation, and enzyme catalysis (isolated enzyme biotransformation). We have absolute strength in the production of HMOs and try our best to meet the needs of clients.

Synthesis of HMOs

So far, there have been many methods to extract or synthesize HMOs, and biosynthetic methods are used to synthesize HMOs because they do not use toxic reagents and are convenient for large-scale production. The research focus of biosynthetic HMOs includes carbon skeleton construction, fucosylation, or sialylation. At present, the mass production of HMO is more focused on the fermentation production of lacto-N-neotetraose (LNnT) and 2'-fucosyllactose (2'FL). For other HMOs, few studies can achieve industrial-scale production synthesis, mainly because of the lack of methods to obtain oligosaccharides with asymmetry or high polymerization, and the limitations in the synthesis of desired oligosaccharide structure.

Fucosidic and sialic acid linkages in HMOs. Fig.1 Fucosidic and sialic acid linkages in HMOs. (Faijes, et al., 2022)

HMO Production by Biocatalytic Methods at CD BioGlyco

Fermentation and isolated enzyme biotransformations are very significant in the production of HMOs, and this class of biocatalytic technologies has made remarkable progress at CD BioGlyco.

  • Whole-cell biotransformation (fermentation) methods
  • Fermentation is carried out by using a system design based on relevant metabolic engineering and metabolic pathways.

    • Using introducing, overexpression, or inactivating microbial genes, etc., the transgenic microorganisms express as much as possible the genes encoding the enzymes involved in the biosynthesis of oligosaccharides.
    • Some HMOs are obtained in the cell factory by using common carbon sources such as glucose, lactose, and glycerin. Through the proper combination, using different strains for mixed fermentation can also optimize the production of HMO.
    • In modified bacteria that can be used in the production of HMOs, the metabolic pathway for synthesizing HMOs is constructed.

    • These include lactic acid bacteria, yeast, and Bacillus subtilis which is generally considered a safe species.
    • Lactic acid bacteria are always used in the fermentation of dairy products.
    • Since milk or yogurt contains a lot of lactose, they can be directly used as fermentation to produce dairy products containing HMOs.
  • Enzyme catalysis (isolated enzyme biotransformation)
  • Enzymatic catalysis requires the isolation and purification of the required enzymes from microorganisms, which are genetically engineered to display the required activity.

    Using the best-given enzyme reaction conditions, add enzyme preparations to the reaction system to obtain HMO.

In terms of fermentation and enzyme catalysis (isolated enzyme biotransformation) technologies, we have extensive experimental experience. Through the discovery of metabolic systems, CD BioGlyco is constantly expanding the product range of HMOs.

The HMOs we synthesize include but are not limited to the species listed in the table below:

HMOs Type of synthesis
2'-Fucosyllactose (2'FL) Fermentation
Enzyme catalysis
3-Fucosyllactose (3FL) Fermentation
Enzyme catalysis
Lacto-N-biose (LNB) Enzyme catalysis
Lacto-N-tetraose (LNT) Fermentation
Enzyme catalysis
Lacto-N-triose (LNT II) Enzyme catalysis
Lacto-N-neotetraose (LNnT) Fermentation
Enzyme catalysis
Lacto-N-fucopentaose I (LNFPI) Fermentation
Enzyme catalysis
Lacto-N-fucopentaose II (LNFPII) Enzyme catalysis
Lacto-N-fucopentaose III (LNFPIII) Enzyme catalysis
Lacto-N-difucohexaose I (LNDFH I) Enzyme catalysis
Lacto-N-difucohexaose II (LNDFH II) Enzyme catalysis
Lacto-N-neodifucohexaose II (LNnDFH II) Enzyme catalysis
3’-Sialyllactose (3’SL) Enzyme catalysis
6’-Sialyllactose (6’SL) Enzyme catalysis

CD BioGlyco has never stopped the continuous exploration and development of new technologies and strategies for HMO production. If you are interested in our services, please feel free to contact us.

References

  1. Pérez-Escalante, E.; et al. Human milk oligosaccharides as bioactive compounds in infant formula: recent advances and trends in synthetic methods. Critical Reviews in Food Science and Nutrition. 2022, 62(1): 181-214.
  2. Faijes, M.; et al. Enzymatic and cell factory approaches to the production of human milk oligosaccharides. Biotechnology advances. 2019, 37(5): 667-697.
This service is for Research Use Only, not intended for any clinical use.

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