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    ARTES heterologous production of holoproteins in Hansenula polymorpha yeast

    products-servicesARTES Biotechnology GmbH
    August 25th 2022

    ARTES Biotechnology Hansenula polymorpha yeast-based production platform is optimised to the production of biologically active and authentic holoproteins, which require either a co-factor or a prosthetic group. The processes have been proven in pilot scale fermentation in animal-free media and without methanol induction.

    This indicates the native potential of the H. polymorpha platform to provide stable and active holoprotein complexes in amounts sufficient for large scale and high level production needed for the growing demand for pharmaceuticals, as well for ethical food production.

    ARTES Hansenula polymorpha protein production platform

    Hansenula polymorpha is a ubiquitous yeast species present in soil, water and on fruits. It is non-pathogenic to animals and humans, does not produce toxins and is an established platform for production of recombinant proteins.

    Examples of products marketed and produced in ARTES ́ Hansenula expression platform are diversified in application. Clients products include e.g. HEPLISAV B by Dynavax, Germany and USA, HEPAVAX-Gene by Berna Biotech Korea Corp., Genevac-B by Serum Institute of India and the HBsAg component of multivalent vaccines (e.g. HEXYON resp HEXACIMA, Sanofi Pasteur, France), Insulin (Wockhardt Ltd., India), Interferon alpha2a and Hirudin (Minapharm Biogenetics, Egypt).

    ARTES has designed its Hansenula host cell line RB11 and its derivatives with one to three auxotrophic marker (ura-, ade-, leu-;), allowing multi-step targeted genetic engineering to improve productivity by increasing integrated expression vector copies and product secretion by chaperone engineering. As a result recombinant products are localized intra- or extra-cellular as soluble, active proteins at high titers of up to 25 g/l.

    For the manufacturing of feed/food products, ARTES has developed its B14 host/vector toolbox, which is free of any antibiotic resistance genes. The fermentation processes are free of antibiotics and of animal derived components. These features and several safety studies provide customers with the basis for large scale manufacturing of enzymes applied in the feed and food industries, such as Hexose oxidase and lipase. H. polymorpha derived products have currently been registered in more than 60 countries worldwide, including FDA approval, and have been recommended by the WHO for mass vaccination programs.

    Holoproteins

    A class of commercially important products are expressed as apoproteins that require a co-factor or a prosthetic group to form a stable and active holoprotein complex.

    Examples of heterologous holoproteins expressed in yeast by recombinant DNA technology have been reported earlier for Saccharomyces cerevisiae1, Pichia pastoris2, and H. polymorpha3.

    A recent ARTES White Paper reports on high-level intracellular expression of different holoproteins including globin proteins.

    For more information or a download, please visit us on www.artes-biotechnology.com.

    References:

    1. Ishchuk OP, Martínez JL, Petranovic D. Improving the Production of Cofactor-Containing Proteins: Production of Human Hemoglobin in Yeast. Methods Mol Biol. 2019;1923:243– 64.
    2. Zhao Xinrui, Yu Fei, Wang Ziwei, Zhou Jingwen, Du Guocheng, Chen Jian, Li Jianghua Construction of Pichia pas-toris recombinant strain for efficiently expressing myoglobin/ hemoglobin from different sources. CN113136349A, 2021
    3. Gellissen G., Piontek M., Dahlem U., Jenzelewski V., Gavagan J.E., DiCosimo R., Anton D.L., Janowicz Z.A. Recombinant Hansenula polymorpha as a biocatalyst: coexpression of the spinach glycolate oxidase (GO) and the S. cerevisiae catalase T (CTT1) gene Appl Microbiol Biotechnol 1996 46:46-54

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