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1. 发明授权

US06168931A Enhanced in vitro synthesis of biological macromolecules using a novel ATP regeneration system 有权
标题翻译：使用新型ATP再生系统增强生物大分子的体外合成
公开(公告)号：US06168931A
公开(公告)日：2001-01-02
申请号：US09270814
申请日：1999-03-17
申请人： James R. Swartz , Dong-Myung Kim
发明人： James R. Swartz , Dong-Myung Kim
IPC分类号： C12P2106
CPC分类号： C12N9/1033 , C07K14/5255 , C12P21/00
摘要： Compositions and methods are provided for the enhanced in vitro synthesis of biological molecules where ATP is required for synthesis. Of particular interest is the synthesis of polymers, e.g. nucleic acids, polypeptides, and complex carbohydrates. A homeostatic system is 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. Since inorganic phosphate can be an inhibitory by-product of synthesis, the period of time when synthesis is maintained in vitro can be extended. 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：提供了用于合成需要ATP的生物分子的增强的体外合成的组合物和方法。特别令人感兴趣的是聚合物的合成，例如核酸，多肽和复合碳水化合物。使用稳态系统来生产ATP，其中所需的高能磷酸键是原位生成的。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。由于无机磷酸盐可以是合成的抑制副产物，因此可以延长合成时间在体外保持的时间。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。

2. 发明申请

US20120077242A1 Efficient Cell-Free Hydrogen Production 有权
标题翻译：高效无细胞氢生产
公开(公告)号：US20120077242A1
公开(公告)日：2012-03-29
申请号：US13246542
申请日：2011-09-27
申请人： James R. Swartz , Phillip Smith
发明人： James R. Swartz , Phillip Smith
IPC分类号： C12P3/00 , C12N9/02
CPC分类号： C12P3/00
摘要： Cell-free synthesis of hydrogen from glucose and cellulosic hydrolysates is provided. Bacterial cells are modified to express high levels of (i) active [FeFe] hydrogenase; (ii) ferredoxin; and (iii) ferredoxin-NADP-reductase (FNR). The cells are then lysed and the lysate is combined with substrate during a production phase, where H2 is produced. The substrate is typically a sugar, e.g. glucose, cellulose hydrolysates, fructose, and the like, including pentose sugars capable of entering the bacterial pentose phosphate cycle. The reaction mixture may be further supplemented with one or more of niacin as a precursor to nicotinamide; a nuclease, particularly a ribonuclease, to break down nucleic acids and generate adenine; and iodoacetamide to inactivate the normal cellular glycolytic pathway and thus maximize conversion yields.
摘要翻译：提供了从葡萄糖和纤维素水解产物中无细胞合成氢气。细菌细胞被修饰以表达高水平的（i）活性[FeFe]氢化酶; （二）铁氧还蛋白; 和（iii）铁氧还蛋白-NADP-还原酶（FNR）。然后将细胞裂解，并在生产阶段中将裂解物与底物结合，其中产生H2。底物通常是糖，例如葡萄糖，纤维素水解物，果糖等，包括能够进入细菌戊糖磷酸循环的戊糖。反应混合物可进一步补充一种或多种作为烟酰胺前体的烟酸; 核酸酶，特别是核糖核酸酶，以分解核酸并产生腺嘌呤; 和碘乙酰胺灭活正常细胞糖酵解途径，从而使转化产率最大化。

3. 发明授权

US07294483B2 Process for producing recombinant polypeptides via a glycerol phosphate or sugar phosphate feed 有权
标题翻译：通过甘油磷酸盐或糖磷酸盐饲料生产重组多肽的方法
公开(公告)号：US07294483B2
公开(公告)日：2007-11-13
申请号：US11077272
申请日：2005-03-10
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12N15/09 , C12P21/02
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。

4. 发明授权

US5639635A Process for bacterial production of polypeptides 失效
标题翻译：细菌生产多肽的方法
公开(公告)号：US5639635A
公开(公告)日：1997-06-17
申请号：US333912
申请日：1994-11-03
申请人： John C. Joly , James R. Swartz
发明人： John C. Joly , James R. Swartz
IPC分类号： C12N15/09 , C07K1/113 , C07K14/245 , C07K14/47 , C07K14/65 , C12N1/21 , C12N9/02 , C12N9/90 , C12N15/70 , C12P21/00 , C12P21/02 , C12R1/19 , C12P21/06
CPC分类号： C12N9/0051 , C07K1/1133 , C07K14/245 , C07K14/65 , C12N15/70 , C12N9/90 , C12P21/02 , C07K2319/00
摘要： A process is provided for producing a heterologous polypeptide in bacteria, which process comprises:(a) culturing bacterial cells, which cells comprise nucleic acid encoding a DsbA or DsbC protein, nucleic acid encoding the heterologous polypeptide, a signal sequence for secretion of both the DsbA or DsbC protein and the heterologous polypeptide, and an inducible promoter for both the nucleic acid encoding the DsbA or DsbC protein and the nucleic acid encoding the heterologous polypeptide, under conditions whereby expression of the nucleic acid encoding the DsbA or DsbC protein is induced prior to induction of the expression of the nucleic acid encoding the heterologous polypeptide, and under conditions whereby either both the heterologous polypeptide and the DsbA or DsbC protein are secreted into the periplasm of the bacteria or the heterologous polypeptide is secreted into the medium in which the bacterial cells are cultured; and(b) recovering the heterologous polypeptide from the periplasm or the culture medium.
摘要翻译：提供了用于在细菌中产生异源多肽的方法，该方法包括：（a）培养细菌细胞，所述细胞包含编码DsbA或DsbC蛋白的核酸，编码异源多肽的核酸，用于分泌两者的信号序列 DsbA或DsbC蛋白和异源多肽，以及用于编码DsbA或DsbC蛋白的核酸和编码异源多肽的核酸的诱导型启动子，其条件是先前诱导编码DsbA或DsbC蛋白的核酸的表达诱导编码异源多肽的核酸的表达，并且在异源多肽和DsbA或DsbC蛋白分泌到细菌周质中的异常条件下，或异源多肽分泌到细菌的培养基中培养细胞; 和（b）从周质或培养基中回收异源多肽。

5. 发明授权

US5342763A Method for producing polypeptide via bacterial fermentation 失效
标题翻译：通过细菌发酵生产多肽的方法
公开(公告)号：US5342763A
公开(公告)日：1994-08-30
申请号：US989844
申请日：1992-11-23
申请人： James R. Swartz
发明人： James R. Swartz
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 , C12D21/00
CPC分类号： C12M41/34 , B01F15/00207 , B01F15/0022 , B01F15/00344 , B01F3/04531 , C07K14/65 , C12M27/02 , C12M41/28 , C12N15/01 , C12N9/0053 , B01F2003/04581 , B01F2003/04673 , B01F2003/04879
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.
摘要翻译：提供了由包含编码该多肽的核酸的细菌宿主细胞的发酵产生感兴趣的多肽的方法。在该方法中，所用宿主细胞具有失活的电子传递链。进一步提供了一种确定特定细菌细胞培养物是否具有大规模发酵时溶解氧不稳定性倾向的方法。

6. 发明授权

US5304472A Method of controlling polypeptide production in bacterial cells 失效
标题翻译：控制细菌细胞中多肽生成的方法
公开(公告)号：US5304472A
公开(公告)日：1994-04-19
申请号：US989845
申请日：1992-11-20
申请人： Steven Bass , James R. Swartz
发明人： Steven Bass , James R. Swartz
IPC分类号： C12N15/09 , A61K39/00 , C07K14/245 , C07K14/65 , C12N1/21 , C12N1/38 , C12N15/12 , C12N15/70 , C12P21/02 , C12R1/19 , C12N1/00 , C12N15/08 , C12N15/31 , C12N15/67
CPC分类号： C07K14/245 , C07K14/65 , C12N1/38 , C12N15/70 , A61K39/00
摘要： Nucleic acid is provided encoding a molecule having certain variations within the phosphate-binding region of native E. coli PstS. Additionally provided are bacterial cells comprising this nucleic acid under control of the native pstS gene promoter, and optionally further comprising nucleic acid encoding a polypeptide of interest under control of the alkaline phosphatase (AP) promoter. Bacterial cells containing both pstS variant nucleic acid and polypeptide nucleic acid are cultured in a medium at a concentration of inorganic phosphate that at all phases of cell growth is above the level at which the cells are starved for phosphate. Alternatively, the cells are cultured under conditions whereby the concentration of inorganic phosphate in the culture medium is controlled during the production period so that the polypeptide is produced under the control of the partially induced AP promoter.
摘要翻译：提供编码在天然大肠杆菌PstS的磷酸结合区域内具有某些变异的分子的核酸。另外提供了包含在天然pstS基因启动子控制下的该核酸的细菌细胞，以及任选地还包含编码在碱性磷酸酶（AP）启动子控制下的感兴趣多肽的核酸。含有pstS变体核酸和多肽核酸两者的细菌细胞在无机磷酸盐的浓度的培养基中培养，在细胞生长的所有阶段都高于细胞饥饿磷酸盐的水平。或者，在生产期间控制培养基中的无机磷酸盐浓度的条件下培养细胞，使得多肽在部分诱导的AP启动子的控制下产生。

7. 发明授权

US4554499A Computer controlled motor 失效
标题翻译：计算机控制电机
公开(公告)号：US4554499A
公开(公告)日：1985-11-19
申请号：US664110
申请日：1984-10-24
申请人： Leonard H. Sherman , James R. Swartz , James M. Williams
发明人： Leonard H. Sherman , James R. Swartz , James M. Williams
IPC分类号： G05D7/06 , H02P7/00 , H02P8/00
CPC分类号： G05D7/0676 , H02P7/00 , Y10T137/0329
摘要： To control a motor, a pulse width modulated signal from a computer is fed to a capacitor which stores each pulse as a voltage proportional to the width of the pulse, the capacitor being reset to zero by a leading edge reset circuit. The voltage on the capacitor is sampled by a sample/hold circuit the output of which is fed to a converter which converts the voltage signal to a frequency signal exponentially related to the voltage, and hence to the pulse width. The converter then powers a driver which drives the motor. The exponential relationship between the pulses and the signal fed to the motor permits a large variationin the frequency signal based on a small variation in pulse width and hence permits accurate control of the motor.
摘要翻译：为了控制电动机，来自计算机的脉冲宽度调制信号被馈送到电容器，其将每个脉冲存储为与脉冲宽度成比例的电压，电容器由前沿复位电路复位为零。电容器上的电压由采样/保持电路采样，该采样/保持电路的输出被馈送到转换器，该转换器将电压信号转换为与电压成指数相关的频率信号，并因此转换成脉冲宽度。然后，转换器为驱动电机的驱动器供电。脉冲和馈送到电机的信号之间的指数关系允许基于脉冲宽度的小变化对频率信号进行大的变化，从而允许电机的精确控制。

8. 发明授权

US07858339B1 Process for bacterial production of polypeptides 有权
标题翻译：细菌生产多肽的方法
公开(公告)号：US07858339B1
公开(公告)日：2010-12-28
申请号：US09422528
申请日：1999-10-21
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12P21/06 , C12N9/00 , C12N9/24 , C12N9/26 , C12N1/20 , C12N15/00 , C07H21/04
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，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。

9. 发明授权

US07341852B2 Methods of decoupling reaction scale and protein synthesis yield in batch mode 有权
标题翻译：分批模式中反应量表和蛋白质合成产率的去耦方法
公开(公告)号：US07341852B2
公开(公告)日：2008-03-11
申请号：US10888145
申请日：2004-07-09
申请人： Alexei M. Voloshin , James R. Swartz
发明人： Alexei M. Voloshin , James R. Swartz
IPC分类号： C12P21/04 , C12P19/23
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：生物大分子在薄膜中体外合成，提供高的表面积/体积比，从而提高放大反应的产率。

10. 发明授权

US06180367B2 Process for bacterial production of polypeptides 有权
公开(公告)号：US06180367B2
公开(公告)日：2001-01-30
申请号：US09422712
申请日：1999-10-21
申请人： Woon-Lam Susan Leung , James R. Swartz
发明人： Woon-Lam Susan Leung , James R. Swartz
IPC分类号： C12P2100
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 and a signal sequence for secretion of the heterologous polypeptide, which nucleic acid encoding the heterologous polypeptide is linked to a another promoter that is inducible, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate.

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