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1. 发明申请

WO02072634A3 RECOMBINANT PROTEINASE K 审中-公开
标题翻译：重组蛋白酶K
公开(公告)号：WO02072634A3
公开(公告)日：2003-03-13
申请号：PCT/EP0201322
申请日：2002-02-08
申请人： ROCHE DIAGNOSTICS GMBH , HOFFMANN LA ROCHE , MUELLER RAINER , THALHOFER JOHANN-PETER , REXER BERNHARD , SCHMUCK RAINER , GEIPEL FRANK , GLASER STEPHAN , SCHOEN HELMUT , MEIER THOMAS , RUDOLPH RAINER , LILIE HAUKE , SCHOTT BJOERN
发明人： MUELLER RAINER , THALHOFER JOHANN-PETER , REXER BERNHARD , SCHMUCK RAINER , GEIPEL FRANK , GLASER STEPHAN , SCHOEN HELMUT , MEIER THOMAS , RUDOLPH RAINER , LILIE HAUKE , SCHOTT BJOERN
IPC分类号： C12N15/09 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12N9/12 , C12N9/58 , C12N15/55 , C07K1/113 , C12N15/70
CPC分类号： C12N9/58 , C07K2319/00 , C12Y304/21064
摘要： The invention relates to recombinant proteinase K and to a method for producing recombinant proteinase K, which is characterised by the following steps: a) transformation of a host cell containing a recombinant nucleic acid that codes for the zymogenic precursor of the proteinase K, b) cultivation of the host cell in such a way that the zymogenic precursor of proteinase K occurs in the form of inclusion bodies in said host cell, c) isolation of the inclusion bodies and re-naturing under conditions, from which the protease part of the zymogenic precursor emerges in its natural conformation, d) activation and purification of the re-natured proteinase K.
摘要翻译：在本发明中，重组蛋白酶K进行说明。此外，公开了用于生产重组蛋白酶K的方法，其特点在于：a）转化的宿主细胞用重组核酸，其编码K中蛋白酶的酶原前体，b）以一种方式培养该宿主细胞，该酶原前体通过蛋白酶K在宿主细胞“包涵体”的形式生产的，c）分离“包涵体”，和复性的条件下，其中在它们的天然构象所产生的酶原前体，D）的活化和纯化，性蛋白酶K的蛋白酶部分，

2. 发明申请

WO02064760A3 EXPRESSION OF RECOMBINANT PROTEINASE K FROM TRITIRACHIUM ALBUM IN YEAST 审中-公开
标题翻译：从酵母Tritirachium专辑重组蛋白酶K表达
公开(公告)号：WO02064760A3
公开(公告)日：2002-12-27
申请号：PCT/EP0201144
申请日：2002-02-05
申请人： ROCHE DIAGNOSTICS GMBH , HOFFMANN LA ROCHE , MUELLER RAINER , THALHOFER JOHANN-PETER , GEIPEL FRANK , GLASER STEPHAN , HOELKE WERNER , SCHOEN HELMUT , KIRSCHBAUM THOMAS
发明人： MUELLER RAINER , THALHOFER JOHANN-PETER , GEIPEL FRANK , GLASER STEPHAN , HOELKE WERNER , SCHOEN HELMUT , KIRSCHBAUM THOMAS
IPC分类号： C12N15/09 , C12N1/19 , C12N9/58 , C12N9/60 , C12N15/55 , C12N15/52 , C12N15/66 , C12N15/67 , C12N15/80 , C12P21/00
CPC分类号： C12N9/58
摘要： The invention relates to a method for the expression in solution of a proteinase K encoding gene in yeast, for example in Pichia pastoris, and subsequent secretion thereof into the culture medium. The invention further relates to a purification method of the heterologously expressed and secreted proteinase K.
摘要翻译：本发明涉及一种方法，用于基因在酵母中的编码该蛋白酶K基因可溶性表达，例如在毕赤酵母，随后分泌进入培养基中。此外，异源的纯化方法表达和分泌的蛋白酶K进行说明。

3. 发明申请

WO2007031187A8 CLEAVAGE OF PRECURSORS OF INSULINS BY A VARIANT OF TRYPSIN 审中-公开
标题翻译：通过一系列TRYPSIN对胰岛素前列腺进行清除
公开(公告)号：WO2007031187A8
公开(公告)日：2007-08-16
申请号：PCT/EP2006008380
申请日：2006-08-26
申请人： SANOFI AVENTIS DEUTSCHLAND , HOFFMANN LA ROCHE , ROCHE DIAGNOSTICS GMBH , GEIPEL FRANK , GLASER STEPHAN , HOH CHRISTOPH , KOPETZKI ERHARD , MUELLER RAINER , ZOCHER FRANK
发明人： GEIPEL FRANK , GLASER STEPHAN , HOH CHRISTOPH , KOPETZKI ERHARD , MUELLER RAINER , ZOCHER FRANK
IPC分类号： C07K14/81
CPC分类号： C07K14/62 , C12N9/6427
摘要： The present invention relates to the production of a variant of recombinant trypsin with increased substrate specificity for arginine versus lysine in non-animal host organisms. Moreover, the present invention relates to a variant of recombinant trypsin and their production. Also provided are use of recombinant porcine pancreatic trypsin variants for cleavage of precursors of insulins, and kits containing the variant of trypsin.
摘要翻译：本发明涉及在非动物宿主生物中生产具有增加的对精氨酸与赖氨酸的底物特异性的重组胰蛋白酶的变体。此外，本发明涉及重组胰蛋白酶的变体及其制备。还提供了重组猪胰胰蛋白酶变体用于切割胰岛素前体的试剂，以及含有胰蛋白酶变体的试剂盒。

4. 发明专利

ES2395232T3 Una composición acuosa estabilizada de alfa-galactosidasa y métodos relacionados con la misma 未知
公开(公告)号：ES2395232T3
公开(公告)日：2013-02-11
申请号：ES10003549
申请日：2010-03-31
申请人： ROCHE DIAGNOSTICS GMBH , HOFFMANN LA ROCHE
发明人： HOELKE WERNER , MUELLER RAINER , SCHMIDT MANFRED , SOBEK HARALD , THALHOFER JOHANN-PETER
IPC分类号： C07K14/435
摘要： Una composición acuosa que comprende una proteína bacteriana con actividad enzimática alfa-galactosidasa,que se caracteriza en que dicha composición contiene arginina a una concentración por encima de 100 mM, enparticular a una concentración de al menos 150 mM, y en la que la conductividad de dicha composición es comomucho 40 mS/cm.

5. 发明专利

ES2384108T3 Fosfatasa alcalina codón-optimizada y su expresión en levadura 未知
公开(公告)号：ES2384108T3
公开(公告)日：2012-06-29
申请号：ES06021699
申请日：2001-07-21
申请人： ROCHE DIAGNOSTICS GMBH , HOFFMANN LA ROCHE
发明人： MUELLER RAINER , THALHOFER JOHANN-PETER , GEIPEL FRANK , HOELKE WERNER , GLASER STEPHAN , ECKSTEIN HELLMUT , KIRSCHBAUM THOMAS , BOMMARIUS BETTINA
IPC分类号： C12N9/16 , C12N15/09 , C12N1/19 , C12N15/53 , C12N15/55 , C12N15/69 , C12N15/81 , C12R1/645
摘要： Cepa huésped transformada que se obtiene al llevar a cabo las etapas (a) clonación de una secuencia genética codón-optimizada que codifica para una fosfatasa alcalina eucariota en un primer vector de expresión con un gen de resistencia a un primer antibiótico, y en un segundo vector de expresión con un gen de resistencia a un segundo antibiótico; seguido de (b) transformación de un huésped de levadura con un primer vector de expresión y la selección de transformantes que tienen integrados en el genoma, por lo menos, una copia del primer vector de expresión con una secuencia genética y el gen de resistencia al primer antibiótico, por medio de su crecimiento sobre un medio de cultivo con una baja concentración del primer antibiótico; seguido de (c) aumento del número de copias de genes del primer vector de expresión mediante transformación múltiple con el primer vector de expresión, en el que la selección se realiza por medio del crecimiento sobre un medio de cultivo con una concentración más elevada del primer antibiótico comparada con la etapa (b); seguido de (d) transformación de clones transformados del huésped con el máximo rendimiento de expresión en fosfatasa alcalina con el segundo vector de expresión y la selección de transformantes que, además de las copias del primer vector de expresión han incluido las copias del segundo vector de expresión con la secuencia genética y el gen resistente al segundo antibiótico, por medio del crecimiento en un medio de cultivo con el segundo antibiótico; seguido de (e) aumento del número de copias de los genes del segundo vector de expresión por medio de la transformación múltiple con el segundo vector de expresión, en el que la selección se realiza por medio del crecimiento sobre un medio de cultivo con una concentración más elevada del segundo antibiótico comparada con la etapa (d); seguido de (f) selección de los clones que tienen integrados de forma estable en el genoma varias copias de la secuencia genética y de los genes de resistencia a los marcadores de selección; y (g) examinación de los clones obtenidos en la etapa (f) en cuanto a su expresión elevada por medio de una prueba de actividad de la fosfatasa alcalina; siendo el rendimiento de expresión de los clones obtenidos en la etapa (f) aumentado por el factor 4 con respecto a la expresión de los clones obtenidos en la etapa (b) .

6. 发明专利

JP2010252793A Stabilized aqueous alpha-galactosidase composition and related method to the same 有权
标题翻译：稳定的水性α-乳糖苷酶组合物及其相关方法
公开(公告)号：JP2010252793A
公开(公告)日：2010-11-11
申请号：JP2010099666
申请日：2010-04-23
申请人： F Hoffmann La Roche Ag , エフ.ホフマン−ラロシュアーゲーF. Hoffmann−La Roche Aktiengesellschaft
发明人： HOELKE WERNER , MUELLER RAINER , SCHMIDT MANFRED , SOBEK HARALD , THALHOFER JOHANN-PETER
IPC分类号： C12N9/96 , C12N9/24 , C12N15/09
CPC分类号： C07K14/435
摘要： PROBLEM TO BE SOLVED: To provide an aqueous composition containing a protein having enzymatic activity of an alpha-galactosidase.
SOLUTION: This method for stabilizing the aqueous composition containing the protein having the enzymatic activity of the alpha-galactosidase is provided. The method for preparing the purified aqueous composition containing the protein having the enzymatic activity of the alpha-galactosidase is also provided.
COPYRIGHT: (C)2011,JPO&INPIT
摘要翻译：待解决的问题：提供含有具有α-半乳糖苷酶的酶活性的蛋白质的含水组合物。解决方案：提供了用于稳定含有具有α-半乳糖苷酶的酶活性的蛋白质的含水组合物的方法。还提供了制备含有具有α-半乳糖苷酶的酶活性的蛋白质的纯化含水组合物的方法。版权所有（C）2011，JPO＆INPIT

7. 发明专利

JP2010046074A Carboxypeptidase b expressed by recombination, and purification of the same 有权
标题翻译：通过重新表达的羧氧化酶B及其相似物的纯化
公开(公告)号：JP2010046074A
公开(公告)日：2010-03-04
申请号：JP2009238318
申请日：2009-10-15
申请人： F Hoffmann La Roche Ag , エフ.ホフマン−ラロシュアーゲーF. Hoffmann−La Roche Aktiengesellschaft
发明人： GLASER STEPHAN , GEIPEL FRANK , KIRSCHBAUM THOMAS , REXER BERNHARD , THALHOFER JOHANN-PETER , MUELLER RAINER , GIESSEL CLAUDIA , ECKSTEIN HELLMUT , WOLF ELVIRA
IPC分类号： C12N9/48 , C12N15/09 , C12N9/98
CPC分类号： C12N9/48 , C07K2319/02 , C07K2319/21
摘要： PROBLEM TO BE SOLVED: To provide a method for producing a protein exhibiting carboxypeptidase B activity from a zymogen.
SOLUTION: A vector comprises a nucleotide sequence encoding a pre-protein consisting of a rat pro-carboxypeptidase B fused with a histidine tag and signal peptide at its N-terminal. The protein without substantially containing the carboxypeptidase B pro-peptide, having the carboxypeptidase B activity and lacking C-terminal tyrosine is obtained by a method for transforming a microorganic host organism by the above vector, culturing it, secreting a histidine-tagged pro-carboxypeptidase B in culturing medium, immobilizing the same on a particle metal chelate-compatible matrix bindable with the histidine tag, releasing the protein having the carboxypeptidase B activity into a liquid phase by a trypsin treatment, and purifying the protein having the carboxypeptidase B activity from the liquid phase.
COPYRIGHT: (C)2010,JPO&INPIT
摘要翻译：待解决的问题：提供从酶原产生显示羧肽酶B活性的蛋白质的方法。解决方案：载体包含编码由组氨酸标签融合的大鼠前羧肽酶B和其N-末端的信号肽组成的前蛋白质的核苷酸序列。通过用上述载体转化微生物宿主生物体的方法，培养它们，分泌组氨酸标记的前羧肽酶，获得具有羧肽酶B活性且缺乏C-末端酪氨酸的基本上不含羧肽酶B前肽的蛋白质 B在培养基中，将其固定在可与组氨酸标签结合的颗粒金属螯合物相容性基质上，通过胰蛋白酶处理将具有羧肽酶B活性的蛋白质释放到液相中，并从其中纯化具有羧肽酶B活性的蛋白质液相。版权所有（C）2010，JPO＆INPIT

8. 发明专利

JP2005168501A Recombinantly expressed carboxypeptidase b and purification thereof 有权
标题翻译：重组表达的羧化酶B及其纯化
公开(公告)号：JP2005168501A
公开(公告)日：2005-06-30
申请号：JP2004350397
申请日：2004-12-02
申请人： F Hoffmann La Roche Ag , エフ.ホフマン−ラロシュアーゲーF. Hoffmann−La Roche Aktiengesellschaft
发明人： GLASER STEPHAN , GEIPEL FRANK , KIRSCHBAUM THOMAS , REXER BERNHARD , THALHOFER JOHANN-PETER , MUELLER RAINER , GIESSEL CLAUDIA , ECKSTEIN HELLMUT , WOLF ELVIRA
IPC分类号： C12N15/09 , C12N9/48
CPC分类号： C12N9/48 , C07K2319/02 , C07K2319/21
摘要： PROBLEM TO BE SOLVED: To provide a method for producing a protein with carboxypeptidase B activity from a zymogen, a method for separating the protein from the zymogen; and to provide a method for separating the protein from the zymogen under a nondenaturing condition. SOLUTION: A method to produce the protein comprising the steps of (a) providing a vector comprising a rat pro-carboxypeptidase B that is fused to a Histidine-tag and a signal peptide; (b) transforming a microbial host organism with the vector; (c) the host organism expresses the pre-protein and secretes the Histidine-tagged pro-carboxypeptidase B; (d) immobilizing the secreted peptidase B on a particulate metal chelate affinity matrix; (e) incubating the matrix in a buffer containing trypsin, releasing the protein with carboxypeptidase B activity into the liquid phase; (f) separating the liquid phase containing the protein from the matrix; and (g) purification of the protein obtained from the liquid phase. COPYRIGHT: (C)2005,JPO&NCIPI
摘要翻译：待解决的问题：提供从酶原产生具有羧肽酶B活性的蛋白质的方法，从酶原分离蛋白质的方法; 并提供在非变性条件下从酶原分离蛋白质的方法。解决方案：一种产生蛋白质的方法，包括以下步骤：（a）提供包含与组氨酸标签和信号肽融合的大鼠前羧肽酶B的载体; （b）用载体转化微生物宿主生物; （c）宿主生物表达前蛋白并分泌组氨酸标记的前羧肽酶B; （d）将分泌的肽酶B固定在金属螯合亲和基质颗粒上; （e）在含有胰蛋白酶的缓冲液中培养基质，将羧肽酶B活性释放到液相中; （f）将含有蛋白质的液相与基质分离; 和（g）从液相中获得的蛋白质的纯化。版权所有（C）2005，JPO＆NCIPI

9. 发明专利

JP2011224002A New t7 rna polymerase variant with enhanced thermostability 有权
标题翻译：新型T7 RNA聚合酶具有增强的热稳定性
公开(公告)号：JP2011224002A
公开(公告)日：2011-11-10
申请号：JP2011090825
申请日：2011-04-15
申请人： F Hoffmann La Roche Ag , エフ.ホフマン−ラロシュアーゲーF. Hoffmann−La Roche Aktiengesellschaft
发明人： GREIF MICHAEL , MUELLER RAINER , RUDOLPH CHRISTIAN , RUF ARMIN , SCHMIDT MANFRED , SOBEK HARALD , THALHOFER JOHANN-PETER
IPC分类号： C12N15/09 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12N9/12 , C12P19/34
CPC分类号： C12N9/1247 , C12P19/34 , C12Y207/07006
摘要： PROBLEM TO BE SOLVED: To provide an improved variant of T7 RNA polymerase by introducing new mutation leading to improved thermostability of the enzyme.SOLUTION: There is provided a variant of T7 RNA polymerase leading to improved thermostability by introducing a new mutation at positions Val426, Ser633, Val650, Thr654, Ala702, Val795 and combinations thereof.

10. 发明专利

JP2004201688A Heat-labile desoxyribonculease i variants 有权
公开(公告)号：JP2004201688A
公开(公告)日：2004-07-22
申请号：JP2003422697
申请日：2003-12-19
申请人： F Hoffmann La Roche Ag , エフ.ホフマン−ラロシュアーゲーF. Hoffmann−La Roche Aktiengesellschaft
发明人： MUELLER RAINER , KIRSCHBAUM THOMAS , SUPPMANN BERNHARD , SCHOEN HELMUT , ENGH RICHARD , HOFFMANN ARTUR , THALHOFER JOHANN-PETER , SIEDEL JOACHIM , ENGEL WOLF-DIETER
IPC分类号： C12N15/09 , C12N1/16 , C12N1/19 , C12N9/16 , C12N9/22 , C12N15/55 , C12N15/63 , C12Q1/34 , C12R1/84
CPC分类号： C12N9/22
摘要： PROBLEM TO BE SOLVED: To provide desoxyribonuclease with increased thermolability compared to bovine pancreatic desoxyribonuclease I, and to provide a method with a high cost-efficiency, for producing the desoxyribonuclease by non-animal host organisms. SOLUTION: The bovine pancreatic desoxyribonuclease I variant has a specific amino acid residue of the bovine pancreatic desoxyribonuclease I substituted with a different amino acid, and has activity of the desoxyribonuclease I. The method for producing the same uses a base sequence encoding the variant. A Pichia pastoris strain encodes a preprotein comprising the variant and a signal peptide, and contains a Pichia pastoris AOX1 promoter or a specific base sequence connected to the promoter element, wherein the base sequence encoding the preprotein contains a chromosome obtained by carrying out fusion of the base sequence of the bovine pancreatic desoxyribonuclease I. COPYRIGHT: (C)2004,JPO&NCIPI

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