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    METABOLIC ENGINEERING OF A GALACTOSE ASSIMILATION PATHWAY IN THE GLYCOENGINEERED YEAST PICHIA PASTORIS
    34.
    发明申请
    METABOLIC ENGINEERING OF A GALACTOSE ASSIMILATION PATHWAY IN THE GLYCOENGINEERED YEAST PICHIA PASTORIS 审中-公开
    葡萄糖生物化学烟草中糖基化途径的代谢工程

    公开(公告)号：US20120003695A1

    公开(公告)日：2012-01-05

    申请号：US13202002

    申请日：2010-02-24

    申请人： Robert C. Davidson ,  Piotr Bobrowicz ,  Dongxing Zha

    发明人： Robert C. Davidson ,  Piotr Bobrowicz ,  Dongxing Zha

    IPC分类号： C12P21/00 ,  C12N9/64 ,  C12N15/81 ,  C12N9/16 ,  C12N9/24 ,  C12N1/19 ,  C12N9/72

    CPC分类号： C12P21/005 ,  C07K16/32 ,  C07K2317/14 ,  C07K2317/41 ,  C12N1/16 ,  C12N9/1205 ,  C12N9/1241 ,  C12N9/90 ,  C12N15/815

    摘要： Lower eukaryotic cells such as Pichia pastoris that normally cannot use galactose as a carbon source but which have been genetically engineered according to the methods herein to use galactose as a sole source of carbon are described. The cells are genetically engineered to express several of the enzymes comprising the Leloir pathway. In particular, the cells are genetically engineered to express a galactokinase, a UDP-galactose-C4-epimerase, and a galactose-1-phosphate uridyltransferase, and optionally a galactose permease. In addition, a method is provided for improving the yield of glycoproteins that have galactose-terminated or -containing N-glycans in cells that have been genetically engineered to produce glycoproteins with N-glycans having galactose residues but which normally lack the enzymes comprising the Leloir pathway comprising transforming the cells with one or more nucleic acid molecules encoding a galactokinase, a UDP-galactose-C4-epimerase, and a galactose-1-phosphate uridyltransferase. The methods and host cells described enable the presence or lack of the ability to assimilate galactose as a selection method for making recombinant cells. The methods and host cells are shown herein to be particularly useful for making immunoglobulins and the like that have galactose-terminated or containing N-glycans.

    摘要翻译： 描述了通常不能使用半乳糖作为碳源但已经根据本文的方法进行遗传工程化的低等真核细胞，例如巴斯德毕赤酵母，其使用半乳糖作为唯一的碳源。 细胞被遗传工程化以表达几种包含Leloir途径的酶。 特别地，细胞被遗传工程化以表达半乳糖激酶，UDP-半乳糖-C4差向异构酶和半乳糖-1-磷酸尿苷酸转移酶，以及任选的半乳糖通透酶。 此外，提供了一种用于提高具有半乳糖封端或含有N-聚糖的糖蛋白的产率的方法，所述糖蛋白在已被遗传工程化以产生具有半乳糖残基的N-聚糖的糖蛋白的细胞中，但通常缺少包含Leloir的酶 途径包括用编码半乳糖激酶，UDP-半乳糖-C4-差向异构酶和半乳糖-1-磷酸尿苷转移酶的一种或多种核酸分子转化细胞。 所描述的方法和宿主细胞使得存在或缺乏同化半乳糖的能力作为制备重组细胞的选择方法。 本文显示的方法和宿主细胞特别可用于制备具有半乳糖终止或含有N-聚糖的免疫球蛋白等。

    PRODUCTION OF GLYCOPROTEINS WITH REDUCED O-GLYCOSYLATION
    36.
    发明申请
    PRODUCTION OF GLYCOPROTEINS WITH REDUCED O-GLYCOSYLATION 有权
    生产具有减少的O-糖苷化的糖蛋白

    公开(公告)号：US20120322101A1

    公开(公告)日：2012-12-20

    申请号：US13530768

    申请日：2012-06-22

    申请人： Piotr Bobrowicz ,  W. James Cook ,  Warren Kett

    发明人： Piotr Bobrowicz ,  W. James Cook ,  Warren Kett

    IPC分类号： C12P21/00

    CPC分类号： C07K16/32 ,  C07K2317/41 ,  C12P21/005

    摘要： A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more α-1,2-mannosidases.

    摘要翻译： 描述了一种用于生产具有减少量的O-连接糖基化的蛋白质组合物的方法。 该方法包括在存在Pmt介导的O-连接糖基化的抑制剂和/或在一种或多种α-1,2-甘露糖苷酶的存在下培养的细胞中产生蛋白质。

    Methods for reducing or eliminating alpha-mannosidase resistant glycans in the production of glycoproteins
    40.
    发明申请
    Methods for reducing or eliminating alpha-mannosidase resistant glycans in the production of glycoproteins 有权
    在糖蛋白生产中减少或消除α-甘露糖苷酶抗性聚糖的方法

    公开(公告)号：US20090093020A1

    公开(公告)日：2009-04-09

    申请号：US12217704

    申请日：2008-07-08

    申请人： Piotr Bobrowicz

    发明人： Piotr Bobrowicz

    IPC分类号： C12N1/19 ,  C12P21/00

    CPC分类号： C07K14/39 ,  C12N9/1051 ,  C12N15/80 ,  C12P21/005

    摘要： The present invention provides methods to reduce or eliminate α-mannosidase resistant glycans on glycoproteins in yeast. The reduction or elimination of α-mannosidase resistant glycans on glycoproteins results from the disruption of the newly isolated P. pastoris AMR2 gene encoding β1,2-mannosyltransferase. The present invention also discloses novel genes, polypeptides, antibodies, vectors and host cells relating to α-mannosidase resistance on glycans.

    摘要翻译： 本发明提供了减少或消除酵母中糖蛋白的α-甘露糖苷酶抗性聚糖的方法。 糖蛋白上的α-甘露糖苷酶抗性聚糖的还原或消除是由新分离的编码β1,2-甘露糖基转移酶的巴斯德毕赤酵母AMR2基因的破坏引起的。 本发明还公开了与聚糖上的α-甘露糖苷酶抗性有关的新基因，多肽，抗体，载体和宿主细胞。