公开(公告)号：US20140212923A1

公开(公告)日：2014-07-31

申请号：US14241505

申请日：2012-08-31

申请人： Roland Weis ,  Thomas Purkarthofer

发明人： Roland Weis ,  Thomas Purkarthofer

IPC分类号： C12P21/00

CPC分类号： C12P21/00 ,  C07K14/55 ,  C07K14/765 ,  C07K14/79 ,  C12N9/0006 ,  C12N9/90 ,  C12N15/815 ,  C12P21/02 ,  C12Y101/03013 ,  C12Y503/04001

摘要： The present invention relates to a genetically modified yeast cell comprising: —at least one recombinant promoter operably linked to at least one gene encoding a polypeptide or protein supporting the biosynthesis of polypeptides or proteins within said cell, said at least one gene being located at the native genomic locus of the genetically unmodified wild-type yeast cell, wherein the naturally occurring promoter of the at least one gene encoding the biosynthesis supporting polypeptide or protein is inactivated by at least one mutation within said naturally occurring promoter and, —a secretion cassette comprising a recombinant nucleic acid molecule encoding a protein or polypeptide of interest and a method for producing a recombinant protein or polypeptide of interest using such a cell.

摘要翻译： 本发明涉及遗传修饰的酵母细胞，其包含：至少一个重组启动子，其可操作地连接至编码支持所述细胞内的多肽或蛋白质的生物合成的多肽或蛋白质的至少一个基因，所述至少一个基因位于 遗传性未修饰的野生型酵母细胞的天然基因组座位，其中编码所述生物合成支持多肽或蛋白质的所述至少一个基因的天然存在的启动子通过所述天然存在的启动子内的至少一个突变而失活，以及包含 编码感兴趣的蛋白质或多肽的重组核酸分子和使用这种细胞产生感兴趣的重组蛋白质或多肽的方法。

公开(公告)号：US09206454B2

公开(公告)日：2015-12-08

申请号：US14241505

申请日：2012-08-31

申请人： Roland Weis ,  Thomas Purkarthofer

发明人： Roland Weis ,  Thomas Purkarthofer

IPC分类号： C12P21/02 ,  C12P21/00 ,  C07K14/55 ,  C07K14/765 ,  C07K14/79 ,  C12N9/04 ,  C12N9/90 ,  C12N15/81

CPC分类号： C12P21/00 ,  C07K14/55 ,  C07K14/765 ,  C07K14/79 ,  C12N9/0006 ,  C12N9/90 ,  C12N15/815 ,  C12P21/02 ,  C12Y101/03013 ,  C12Y503/04001

摘要： The present invention relates to a genetically modified yeast cell comprising: —at least one recombinant promoter operably linked to at least one gene encoding a polypeptide or protein supporting the biosynthesis of polypeptides or proteins within said cell, said at least one gene being located at the native genomic locus of the genetically unmodified wild-type yeast cell, wherein the naturally occurring promoter of the at least one gene encoding the biosynthesis supporting polypeptide or protein is inactivated by at least one mutation within said naturally occurring promoter and, —a secretion cassette comprising a recombinant nucleic acid molecule encoding a protein or polypeptide of interest and a method for producing a recombinant protein or polypeptide of interest using such a cell.

摘要翻译： 本发明涉及遗传修饰的酵母细胞，其包含：至少一个重组启动子，其可操作地连接至编码支持所述细胞内的多肽或蛋白质的生物合成的多肽或蛋白质的至少一个基因，所述至少一个基因位于 遗传性未修饰的野生型酵母细胞的天然基因组座位，其中编码所述生物合成支持多肽或蛋白质的所述至少一个基因的天然存在的启动子通过所述天然存在的启动子内的至少一个突变而失活，以及包含 编码感兴趣的蛋白质或多肽的重组核酸分子和使用这种细胞产生感兴趣的重组蛋白质或多肽的方法。

公开(公告)号：US20100041110A1

公开(公告)日：2010-02-18

申请号：US11795685

申请日：2005-12-28

申请人： Roland Weis ,  Anton Glieder ,  Karl Gruber ,  Wolfgang Skrang ,  Oliver Maurer ,  Richard Gaisberger

发明人： Roland Weis ,  Anton Glieder ,  Karl Gruber ,  Wolfgang Skrang ,  Oliver Maurer ,  Richard Gaisberger

IPC分类号： C12P13/00 ,  C12N9/88

CPC分类号： C12P13/004 ,  C12N9/88

摘要： R-hydroxynitrile lyases having an improved substrate acceptance, increased activity and increased selectivity, in which there is replacement in the amino acid sequence of R-hydroxynitrile lyases from the Rosaceae family either a) of the amino acid residue which corresponds to position 360 of the mature PaHNL5 protein by another apolar amino acid or a neutral amino acid and/or b) of the amino acid residue which corresponds to position 225 of the mature PaHNL5 protein by another polar amino acid, it also being possible where appropriate for 1 to 20 further residues in the active center or in the hydrophobic channel leading to the active center to be replaced.

摘要翻译： 具有改善的底物接受性，增加的活性和增加的选择性的R-羟基腈裂解酶，其中在来自蔷薇科家族的R-羟基腈裂解酶的氨基酸序列中置换a）的氨基酸残基，其对应于 通过另一个非极性氨基酸或中性氨基酸的成熟PaHNL5蛋白和/或b）与另一种极性氨基酸对应的成熟PaHNL5蛋白的位置225的氨基酸残基，在适当的情况下还可以进一步加入1至20 活性中心或疏水通道中的残留物，导致活性中心被更换。

公开(公告)号：US20060105434A1

公开(公告)日：2006-05-18

申请号：US10548271

申请日：2004-02-24

申请人： Wolfgang Skranc ,  Anton Glieder ,  Karl Gruber ,  Roland Weis ,  Ruud Luiten

发明人： Wolfgang Skranc ,  Anton Glieder ,  Karl Gruber ,  Roland Weis ,  Ruud Luiten

IPC分类号： C12P13/04 ,  C12P13/00 ,  C12N9/88

CPC分类号： C12P13/004 ,  C12N9/88

摘要： The invention related to R-hydroxynitrillyases from the family of Rosaceae that are characterized by an improved substrate tolerance and increased stability. In the active center of the R-hydroxynitrillyases either a) an alanine group is substituted by glycine, valine, leucine, isoleucine, or phenylalanine or b) a phenylalanine group is substituted by alanine, glycine valine, leucine or isoleucine, or c) a leucine group is substituted by alanine, glycine, valine, isoleucine or phenylalanine, or d) an isoleucine group is substituted by alanine, glycine, valine, leucine or phenylalanine. The invention also relates to the use of these lyases in the production of enantiomer-pure R- or S-cyanohydrines.

摘要翻译： 本发明涉及来自蔷薇科家族的R-羟基亚硝酸酯酶，其特征在于改善的底物耐受性和增加的稳定性。 或者a）丙氨酸被甘氨酸，缬氨酸，亮氨酸，异亮氨酸或苯丙氨酸取代，或b）苯丙氨酸被丙氨酸，甘氨酸缬氨酸，亮氨酸或异亮氨酸取代，或c） 亮氨酸被丙氨酸，甘氨酸，缬氨酸，异亮氨酸或苯丙氨酸取代，或d）异亮氨酸被丙氨酸，甘氨酸，缬氨酸，亮氨酸或苯丙氨酸取代。 本发明还涉及这些裂解酶在制备对映异构体纯的R-或S-氰基中的用途。

公开(公告)号：US07572608B2

公开(公告)日：2009-08-11

申请号：US10548271

申请日：2004-02-24

申请人： Wolfgang Skranc ,  Anton Glieder ,  Karl Gruber ,  Roland Weis ,  Ruud Luiten

发明人： Wolfgang Skranc ,  Anton Glieder ,  Karl Gruber ,  Roland Weis ,  Ruud Luiten

IPC分类号： C12P13/04 ,  C12N9/88

CPC分类号： C12P13/004 ,  C12N9/88

摘要： The invention relates to R-hydroxynitrillyases from the family of Rosaceae that are characterized by an improved substrate tolerance and increased stability. In the active center of the R-hydroxynitrillyases either a) an alanine group is substituted by glycine, valine, leucine, isoleucine, or phenylalanine or b) a phenylalanine group is substituted by alanine, glycine valine, leucine or isoleucine, or c) a leucine group is substituted by alanine, glycine, valine, isoleucine or phenylalanine, or d) an isoleucine group is substituted by alanine, glycine, valine, leucine or phenylalanine. The invention also relates to the use of these lyases in the production of enantiomer-pure R- or S-cyanohydrines.

摘要翻译： 本发明涉及来自蔷薇科的R-羟基亚硝酸酯酶，其特征在于改善的基质耐受性和增加的稳定性。 或者a）丙氨酸被甘氨酸，缬氨酸，亮氨酸，异亮氨酸或苯丙氨酸取代，或b）苯丙氨酸被丙氨酸，甘氨酸缬氨酸，亮氨酸或异亮氨酸取代，或c） 亮氨酸被丙氨酸，甘氨酸，缬氨酸，异亮氨酸或苯丙氨酸取代，或d）异亮氨酸被丙氨酸，甘氨酸，缬氨酸，亮氨酸或苯丙氨酸取代。 本发明还涉及这些裂解酶在制备对映异构体纯的R-或S-氰基中的用途。