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    O-Mannosyltransferase Deficient Filamentous Fungal Cells and Methods of Use Thereof
    2.
    发明申请
    O-Mannosyltransferase Deficient Filamentous Fungal Cells and Methods of Use Thereof 审中-公开
    O-甲基转移酶缺陷型丝状真菌细胞及其使用方法

    公开(公告)号：US20160376570A1

    公开(公告)日：2016-12-29

    申请号：US14902492

    申请日：2014-07-03

    申请人： NOVARTIS AG ,  GLYKOS FINLAND OY

    发明人： Jari Natunen ,  Jukka Hiltunen ,  Anne Huuskonen ,  Markku Saloheimo ,  Christian Ostermeier ,  Benjamin Patrick Sommer ,  Ramon Wahl

    IPC分类号： C12N9/10 ,  C07K16/00

    CPC分类号： C12N9/1051 ,  C07K14/37 ,  C07K16/00 ,  C07K16/18 ,  C07K2317/41 ,  C12Y204/01109

    摘要： The present disclosure relates to compositions and methods useful for the production of heterologous proteins with reduced O-mannosylation in filamentous fungal cells, such as Trichoderma cells. More specifically, the invention provides a PMT-deficient filamentous fungal cell comprising a) at least a first mutation that reduces an endogenous protease activity compared to a parental filamentous fungal cell which does not have said first mutation, and, b) at least a second mutation in a PMT gene that reduces endogenous O-mannosyltransferase activity compared to a parental filamentous fungal cell which does not have said second mutation, wherein said filamentous fungal cell is selected from the group consisting of Trichoderma, Neurospora, Myceliophthora or Chrysosporium cell.

    摘要翻译： 本公开涉及可用于生产丝状真菌细胞如木霉属细胞中具有降低的O-甘露糖基化的异源蛋白质的组合物和方法。 更具体地，本发明提供了一种缺乏PMT的丝状真菌细胞，其包含a）与不具有所述第一突变的亲本丝状真菌细胞相比，降低内源性蛋白酶活性的至少第一突变，和b）至少第二突变 与不具有第二突变的亲本丝状真菌细胞相比，降低内源性O-甘露糖基转移酶活性的PMT基因中的突变，其中所述丝状真菌细胞选自木霉属，孢子孢子虫，丝霉菌属或孢子孢子菌属细菌。

    CONTROLLING O-GLYCOSYLATION IN LOWER EUKARYOTES
    4.
    发明申请
    CONTROLLING O-GLYCOSYLATION IN LOWER EUKARYOTES 审中-公开
    控制低蛋白质中的O-糖苷化

    公开(公告)号：US20140302556A1

    公开(公告)日：2014-10-09

    申请号：US14354144

    申请日：2012-10-22

    申请人： MERCK SHARP & DOHEME CORP.

    发明人： Bo Jiang ,  Stephanie A. Nelson ,  Rebecca D. Argyros ,  Dongxing Zha

    IPC分类号： C12N15/81 ,  C12P21/00

    CPC分类号： C12N15/81 ,  C07K16/1027 ,  C07K16/32 ,  C07K2317/41 ,  C12N9/1051 ,  C12P21/00 ,  C12P21/005 ,  C12Y204/01109

    摘要： Lower eukaryote host cells in which expression of the endogenous protein mannosyltransferase 2 (PMT2) gene has been disrupted by introducing a nucleic acid molecule encoding a Pmt2p protein having a mutation in a conserved region of the protein. The mutation confers to the host cell resistance to PMT inhibitors, which are used to reduce the amount of O-glycosylation of recombinant proteins produced by the host cells but which also have the effect of reducing the robustness of the host cells during fermentation. When host cells that express the mutated PMT2 gene but not the endogenous Pmt2p are cultivated in the presence of a P MT inhibitor, the host cells display an increase in cellular robustness during fed-batch fermentation and express recombinant pro teins in high yield while the amounts O-glycosylation are similar to that produced under similar conditions by host cells that express only the endogenous P MT2 gene.

    摘要翻译： 通过引入编码在蛋白质的保守区域中具有突变的Pmt2p蛋白质的核酸分子已经破坏内源性蛋白质甘露糖基转移酶2（PMT2）基因的表达的下位真核生物宿主细胞。 该突变赋予宿主细胞对PMT抑制剂的抗性，其用于减少宿主细胞产生的重组蛋白质的O-糖基化量，但也具有降低发酵过程中宿主细胞稳健性的作用。 当在P MT抑制剂的存在下培养表达突变的PMT2基因而不是内源性Pmt2p的宿主细胞时，在补料分批发酵期间宿主细胞表现出细胞稳健性的增加，并以高产量表达重组蛋白， O-糖基化与在仅表达内源性P MT2基因的宿主细胞在相似条件下产生的相似。