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1. 发明申请

WO2005087802A3 PROCESS FOR PRODUCING POLYPEPTIDES 审中-公开
标题翻译：生产多糖的方法
公开(公告)号：WO2005087802A3
公开(公告)日：2006-03-02
申请号：PCT/US2005007880
申请日：2005-03-10
申请人： GENENTECH INC , WOON-LAM SUSAN LEUNG , SWARTZ JAMES R
发明人： WOON-LAM SUSAN LEUNG , SWARTZ JAMES R
IPC分类号： C07H21/04 , C07K14/47 , C07K14/65 , C07K14/705 , C07K16/00 , C07K16/26 , C12N1/20 , C12N1/21 , C12N1/32 , C12N1/38 , C12N15/70 , C12N15/74 , C12P21/00 , C12P21/02 , C12P21/06
CPC分类号： C12N15/67 , C07K16/26 , C07K2317/22 , C12N1/20 , C12N1/32 , C12N1/38 , C12N15/70 , C12P21/00
摘要： A process is described for producing a polypeptide heterologous to E. coli wherein E. coli cells comprising nucleic acid encoding the polypeptide are cultured in a culture medium while feeding to the culture medium a transportable organophosphate, such that the nucleic acid is expressed. The polypeptide is then recovered from the cells.
摘要翻译：描述了用于产生与大肠杆菌异源的多肽的方法，其中包含编码多肽的核酸的大肠杆菌细胞在培养基中培养，同时向培养基中加入可运输的有机磷酸酯，使得表达核酸。然后从细胞中回收多肽。

2. 发明申请

WO9856930A9 METHODS FOR PRODUCING HETEROLOGOUS DISULFIDE BOND-CONTAINING POLYPEPTIDES IN BACTERIAL CELLS 审中-公开
标题翻译：在细菌细胞中生产含异源二硫键的多肽的方法
公开(公告)号：WO9856930A9
公开(公告)日：1999-05-06
申请号：PCT/US9812004
申请日：1998-06-09
申请人： UNIV TEXAS , GENENTECH INC , GEORGIOU GEORGE , QIU JI , BESSETTE PAUL , SWARTZ JAMES R
发明人： GEORGIOU GEORGE , QIU JI , BESSETTE PAUL , SWARTZ JAMES R
IPC分类号： C12N15/09 , C07K14/47 , C07K14/65 , C07K14/81 , C12N1/21 , C12N9/02 , C12N9/64 , C12N9/72 , C12N15/70 , C12P21/02
CPC分类号： C12N9/0051 , C07K14/4743 , C07K14/8117 , C12N9/6459 , C12N15/70 , C12Y304/21069
摘要： Disclosed are methods and compositions for producing heterologous disulfide bond containing polypeptides in bacterial cells. In preferred embodiments, the methods involve co-expression of a prokaryotic disulfide isomerase, such as DsbC or DsbG and a gene encoding a recombinant eukaryotic polypeptide. Exemplary polypeptides disclosed include tissue plasminogen activator.
摘要翻译：公开了用于在细菌细胞中产生含有异源二硫键的多肽的方法和组合物。在优选的实施方案中，所述方法涉及原核二硫化物异构酶（例如DsbC或DsbG）和编码重组真核生物多肽的基因的共表达。所公开的示例性多肽包括组织纤溶酶原激活物。

3. 发明申请

WO9412630A3 METHOD FOR PRODUCING POLYPEPTIDE VIA BACTERIAL FERMENTATION 审中-公开
标题翻译：通过生物发酵生产多肽的方法
公开(公告)号：WO9412630A3
公开(公告)日：1994-10-13
申请号：PCT/US9311297
申请日：1993-11-19
申请人： GENENTECH INC , SWARTZ JAMES R
发明人： SWARTZ JAMES R
IPC分类号： C12N15/09 , B01F3/04 , B01F15/00 , C07K14/65 , C12M1/00 , C12M1/06 , C12M1/36 , C12M3/02 , C12N1/21 , C12N9/02 , C12N15/01 , C12P21/02 , C12Q1/02
CPC分类号： C12M41/34 , B01F3/04531 , B01F15/00207 , B01F15/0022 , B01F15/00344 , B01F2003/04581 , B01F2003/04673 , B01F2003/04879 , C07K14/65 , C12M27/02 , C12M41/28 , C12N9/0053 , C12N15/01
摘要： A process for producing a polypeptide of interest from fermentation of bacterial host cells comprising nucleic acid encoding the polypeptide is provided. In this method, the host cells employed have an inactivated electron transport chain. Further provided is a method for determining if a particular bacterial cell culture has a propensity for dissolved oxygen instability when fermented on a large scale.

4. 发明申请

WO0055353A9 IN VITRO MACROMOLECULE BIOSYNTHESIS METHODS USING EXOGENOUS AMINO ACIDS AND A NOVEL ATP REGENERATION SYSTEM 审中-公开
标题翻译：在体外大分子生物化学方法中使用异构氨基酸和新型ATP再生系统
公开(公告)号：WO0055353A9
公开(公告)日：2002-03-14
申请号：PCT/US0007095
申请日：2000-03-15
申请人： UNIV LELAND STANFORD JUNIOR , SWARTZ JAMES R , KIM DONG MYUNG
发明人： SWARTZ JAMES R , KIM DONG-MYUNG
IPC分类号： C12N15/09 , C07K1/02 , C07K14/525 , C12N9/10 , C12P21/00 , C12P21/02 , C12P21/06
CPC分类号： C12N9/1033 , C07K14/5255 , C12P21/00
摘要： Compositions and methods are provided for the enhanced in vitro synthesis of polypeptides. In order to improve the performance of in vitro protein synthesis reactions, metabolic inhibitors, or manipulation of a source organism, is used to diminish or avoid the action of enzymes responsible for undesirable amino acids production or depletion. A homeostatic system may be used for production of ATP, where the required high energy phosphate bonds are generated in situ, e.g. through coupling with an oxidation reaction. The homeostatic energy source will typically lack high energy phosphate bonds itself, and will therefore utilize free phosphate in the reaction mix during generation of ATP. The homeostatic energy source is provided in combination with an enzyme that catalyzes the creation of high energy phosphate bonds and with an enzyme that can use that high energy phosphate bond to regenerate ATP.
摘要翻译：提供组合物和方法用于增强多肽的体外合成。为了改善体外蛋白质合成反应的性能，使用代谢抑制剂或来源生物体的操作来减少或避免负责不良氨基酸产生或消耗的酶的作用。可以使用稳态系统来生产ATP，其中所需的高能磷酸键是原位产生的。通过与氧化反应的偶联。稳态能源本身通常缺乏高能量磷酸键，因此在产生ATP期间将在反应混合物中利用游离磷酸盐。稳态能量源与催化高能磷酸键的酶的结合，以及能够使用高能磷酸键重新生成ATP的酶组合提供。

5. 发明申请

WO2011140516A3 METHODS FOR CONTROL OF FLUX IN METABOLIC PATHWAYS THROUGH ENZYME RELOCATION 审中-公开
标题翻译：通过酶转移控制代谢途径中的通量的方法
公开(公告)号：WO2011140516A3
公开(公告)日：2012-01-05
申请号：PCT/US2011035639
申请日：2011-05-06
申请人： GREENLIGHT BIOSCIENCES INC , UNIV LELAND STANFORD JUNIOR , SWARTZ JAMES R
发明人： SWARTZ JAMES R
IPC分类号： C12N1/00 , C12N1/06 , C12N15/52
CPC分类号： C12P7/42 , C07K2319/034 , C12N9/1029 , C12N9/88 , C12N15/625 , C12N15/70 , C12P5/007 , C12P7/16 , C12P7/625 , C12P23/00 , Y02E50/10
摘要： Genetically manipulated cells, lysates of such cells, systems, and methods of use thereof are provided, where one or more enzymes in a pathway of interest are genetically modified to incorporate a peptide sequence that provides for relocation of the protein, e.g., to the periplasm, so as to sequester the enzyme, and where the enzyme controls flux in the pathway of interest.
摘要翻译：提供了遗传操纵的细胞，这种细胞的裂解物，系统及其使用方法，其中感兴趣途径中的一种或多种酶被遗传修饰以掺入提供蛋白重新定位的肽序列，例如提供给周质，以便螯合酶，以及酶控制感兴趣途径中的通量。

6. 发明申请

WO2005010155A3 METHODS OF DECOUPLING REACTION SCALE AND PROTEIN SYNTHESIS YIELD IN BATCH MODE 审中-公开
标题翻译：分批模式下反应规模和蛋白质合成产量的解耦方法
公开(公告)号：WO2005010155A3
公开(公告)日：2005-04-28
申请号：PCT/US2004022632
申请日：2004-07-12
申请人： UNIV LELAND STANFORD JUNIOR , VOLOSHIN ALEXEI M , SWARTZ JAMES R
发明人： VOLOSHIN ALEXEI M , SWARTZ JAMES R
IPC分类号： C12N20060101 , C12N15/70 , C12P21/02 , C12P21/06
CPC分类号： C12P21/02
摘要： Biological macromolecules are synthesized in vitro in a thin film that provides for high surface area/volume ratio, allowing improved yield in scaled up reactions.
摘要翻译：生物大分子在体外以薄膜形式合成，提供高表面积/体积比，允许在放大的反应中提高产量。

7. 发明申请

WO9856930A2 METHODS FOR PRODUCING HETEROLOGOUS DISULFIDE BOND-CONTAINING POLYPEPTIDES IN BACTERIAL CELLS 审中-公开
标题翻译：在细菌细胞中生产含有异源结合物的多肽的方法
公开(公告)号：WO9856930A2
公开(公告)日：1998-12-17
申请号：PCT/US9812004
申请日：1998-06-09
申请人： UNIV TEXAS , GEORGIOU GEORGE , QIU JI , BESSETTE PAUL , SWARTZ JAMES R
发明人： GEORGIOU GEORGE , QIU JI , BESSETTE PAUL , SWARTZ JAMES R
IPC分类号： C12N15/09 , C07K14/47 , C07K14/65 , C07K14/81 , C12N1/21 , C12N9/02 , C12N9/64 , C12N9/72 , C12N15/70 , C12P21/02
CPC分类号： C12N9/0051 , C07K14/4743 , C07K14/8117 , C12N9/6459 , C12N15/70 , C12Y304/21069
摘要： Disclosed are methods and compositions for producing heterologous disulfide bond containing polypeptides in bacterial cells. In preferred embodiments, the methods involve co-expression of a prokaryotic disulfide isomerase, such as DsbC or DsbG and a gene encoding a recombinant eukaryotic polypeptide. Exemplary polypeptides disclosed include tissue plasminogen activator.
摘要翻译：公开了用于在细菌细胞中产生含异源二硫键的多肽的方法和组合物。在优选的实施方案中，所述方法涉及原核二硫键异构酶如DsbC或DsbG的共表达和编码重组真核多肽的基因。公开的示例性多肽包括组织纤溶酶原激活剂。

8. 发明申请

WO0024873A9 PROCESS FOR RECOVERING HETEROLOGOUS POLYPEPTIDES FROM BACTERIAL CELLS 审中-公开
标题翻译：从细菌回收异源多肽的方法
公开(公告)号：WO0024873A9
公开(公告)日：2000-11-02
申请号：PCT/US9924696
申请日：1999-10-21
申请人： GENENTECH INC
发明人： LEUNG WOON-LAM SUSAN , SWARTZ JAMES R
IPC分类号： C12N15/09 , C07K14/01 , C07K14/65 , C07K16/22 , C07K16/28 , C07K16/40 , C12N1/06 , C12N1/20 , C12N9/22 , C12N9/36 , C12P21/02 , C12P21/08 , C12N15/70
CPC分类号： C12N9/2462 , C07K14/005 , C07K14/65 , C07K16/2845 , C07K2317/24 , C07K2317/54 , C07K2319/02 , C12N1/06 , C12N9/22 , C12N2795/10222 , C12N2795/10322 , C12P21/02
摘要： Processes are described for recovering heterologous polypeptide from bacterial cells, including the periplasm and cytoplasm. One process involves culturing the bacterial cells, which cells comprise nucleic acid encoding phage lysozyme and nucleic acid encoding a protein that displays DNA-digesting activity, wherein these nucleic acids are linked to a first promoter, and nucleic acid encoding the heterologous polypeptide, which nucleic acid is linked to a second promoter, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate. Another process entails culturing bacterial cells that comprise nucleic acid encoding phage lysozyme, gene t, and nucleic acid encoding a protein that displays DNA-digesting activity under the control of a signal sequence for secretion of said DNA-digesting protein, wherein said nucleic acids are linked to one or more promoters, and nucleic acid encoding the heterologous polypeptide, which nucleic acid encoding the heterologous polypeptide is linked to another promoter that is inducible, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate.
摘要翻译：描述了从细菌细胞（包括周质和细胞质）回收异源多肽的方法。一个过程涉及培养细菌细胞，该细胞包含编码噬菌体溶菌酶的核酸和编码显示DNA消化活性的蛋白质的核酸，其中这些核酸与第一启动子和编码异源多肽的核酸连接，该核酸在某些条件下酸与第二种启动子连接以产生肉汤裂解物; 并从肉汤裂解物中回收积累的异源多肽。另一种方法需要培养包含编码噬菌体溶菌酶的核酸，基因t和编码蛋白质的核酸的细菌细胞，所述蛋白质在用于分泌所述DNA消化蛋白质的信号序列的控制下显示DNA消化活性，其中所述核酸是与一种或多种启动子连接的核酸，以及编码异源多肽的核酸，该核酸编码异源多肽与另一种可诱导的启动子连接，以在某些条件下产生肉汤裂解物; 并从肉汤裂解物中回收积累的异源多肽。

9. 发明申请

WO0024769A3 PROCESS FOR RECOVERING HETEROLOGOUS POLYPEPTIDE FROM BACTERIAL REFRACTILE PARTICLES 审中-公开
标题翻译：从细菌回收颗粒中回收异物多肽的方法
公开(公告)号：WO0024769A3
公开(公告)日：2000-08-17
申请号：PCT/US9924830
申请日：1999-10-21
申请人： GENENTECH INC
发明人： LEUNG WOON-LAM SUSAN , SWARTZ JAMES R
IPC分类号： C07K14/52 , C07K14/65 , C12N9/22 , C12N9/36 , C12N15/70 , C12P21/02 , C12N1/06
CPC分类号： C12N9/2462 , C07K14/52 , C07K14/65 , C12N9/22 , C12N15/70 , C12P21/02
摘要： Refractile particles containing a heterologous polypeptide as an insoluble aggregate are recovered from bacterial periplasm. The process involves culturing bacterial cells so as to express nucleic acid encoding phage lysozyme and nucleic acid encoding the heterologous polypeptide under separate promoters, disrupting the cells mechanically to release the phage lysozyme so as to release refractile particles from the bacterial cellular matrix, and recovering the released refractile particles from the periplasm. Chloroform is not used in any step and the recovery step minimizes co-recovery of cellular debris with the released refractile particles.
摘要翻译：从细菌周质中回收含有异源多肽作为不溶性聚集体的易折射粒子。该方法包括培养细菌细胞，以便在单独的启动子下表达编码噬菌体溶菌酶的核酸和编码异源多肽的核酸，机械破坏细胞以释放噬菌体溶菌酶，以便从细菌细胞基质中释放可折射颗粒，并回收从周质释放出可折射粒子。氯仿不用于任何步骤，并且回收步骤最小化释放的折射率颗粒的细胞碎片的共同回收。

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