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1. 发明申请

WO1996005316A1 OVERPRODUCTION AND PURIFICATION OF SOLUBLE PHA SYNTHASE 审中-公开
标题翻译：可溶性PHA合成酶的超导和纯化
公开(公告)号：WO1996005316A1
公开(公告)日：1996-02-22
申请号：PCT/US1995010235
申请日：1995-08-10
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , METABOLIX, INC.
发明人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , METABOLIX, INC. , PEOPLES, Oliver, P. , GERNGROSS, Tillman, U. , SINSKEY, Anthony, J.
IPC分类号： C12N15/52
CPC分类号： C12N9/93 , C12N9/00 , C12N9/0006 , C12N9/1029 , C12N15/52 , C12P7/625
摘要： Purified, soluble recombinant bacterial long chain and short chain PHA synthases are described, with methods and materials for overexpression and purification.
摘要翻译：描述了纯化的可溶性重组细菌长链和短链PHA合成酶，其具有用于过表达和纯化的方法和材料。

2. 发明申请

WO1998004713A1 METHOD FOR CONTROLLING MOLECULAR WEIGHT OF POLYHYDROXYALKANOATES 审中-公开
标题翻译：控制聚羟基烷基酯分子量的方法
公开(公告)号：WO1998004713A1
公开(公告)日：1998-02-05
申请号：PCT/US1997013301
申请日：1997-07-25
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY
发明人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , SNELL, Kristi, D. , HOGAN, Scott, A. , SIM, Sang, Jun , SINSKEY, Anthony, J. , RHA, Chokyun
IPC分类号： C12N15/52
CPC分类号： C12N9/93 , C12P7/625
摘要： A method has been developed for control of molecular weight and molecular weight dispersity during production of polyhydroxyalkanoates in genetically engineered organism by control of the level and time of expression of one or more PHA synthases in the organisms. The method was demonstrated by constructing a synthetic operon for PHA production in E. coli in which the level of PHA synthase activity could be tightly controlled by placement of the synthase behind an inducible promoter. Modulation of the total level of PHA synthase activity in the host cell by varying the concentration of the inducer, isopropyl beta -D-thiogalactoside (IPTG), was found to effect the molecular weight of the polymer produced in the cell. Specifically, high concentrations of synthase activity were found to yield polymers of low molecular weight while low concentrations of synthase activity yielded polymers of higher molecular weight. Polymer molecular weight dispersity is also proportional to the amount of synthase activity, with less dispersity in polyhydroxyalkanoate compositions produced in expression systems with an initial burst of synthase activity, and higher levels of molecular weight dispersity in polyhydroxyalkanoate compositions produced in expression systems with the levels of synthase activity varied during synthesis of the polyhydroxyalkanoate.
摘要翻译：已经开发了一种通过控制生物体内一种或多种PHA合成酶的表达水平和时间来控制在遗传工程生物体内聚羟基链烷酸酯生产过程中的分子量和分子量分散性的方法。该方法通过在大肠杆菌中构建用于PHA生产的合成操纵子来证明，其中通过将合酶置于诱导型启动子后面可以严格控制PHA合酶活性的水平。发现通过改变诱导剂异丙基β-D-硫代半乳糖苷（IPTG）的浓度来调节宿主细胞中PHA合成酶活性的总水平，从而影响细胞中产生的聚合物的分子量。具体地说，发现高浓度的合成酶活性产生低分子量的聚合物，而低浓度的合成酶活性则产生较高分子量的聚合物。聚合物分子量分散度也与合成酶活性的量成比例，在具有合成酶活性的初始爆发的表达系统中产生的聚羟基链烷酸酯组合物的分散性较低，并且在表达系统中产生的聚羟基链烷酸酯组合物中较高水平的分子量分散性具有合成酶活性在聚羟基链烷酸酯的合成过程中变化。

3. 发明申请

WO2009117036A3 BIOACTIVE MOLECULES FROM CO-CULTIVATION OF MICROBES 审中-公开
公开(公告)号：WO2009117036A3
公开(公告)日：2009-09-24
申请号：PCT/US2008/087936
申请日：2008-12-22
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , SINSKEY, Anthony, J. , LESSARD, Philip, A. , KUROSAWA, Kazuhiko
发明人： SINSKEY, Anthony, J. , LESSARD, Philip, A. , KUROSAWA, Kazuhiko
IPC分类号： C07D498/12 , C07D405/12 , C07D498/02 , A61K31/7056
摘要： Certain aspects of the invention relates to antibiotics, as well as pharmaceutically acceptable salts, pro-drugs and/or analogs thereof. Another aspect of the inventions relates to methods of use of said antibiotics.

4. 发明申请

WO2003093406A2 MICROFERMENTORS FOR RAPID SCREENING AND ANALYSIS OF BIOCHEMICAL PROCESSES 审中-公开
标题翻译：微生物快速筛选和生物化学过程分析
公开(公告)号：WO2003093406A2
公开(公告)日：2003-11-13
申请号：PCT/US2003/013479
申请日：2003-05-01
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY
发明人： ZANZOTTO, Andrea , JENSEN, Klavs, F. , SINSKEY, Anthony, J. , SCHMIDT, Martin, A. , LAIBINIS, Paul , RAM, Rajeev, J. , SZITA, Nicolas
IPC分类号： C12M
CPC分类号： G01N21/648 , B01J2219/00283 , B01J2219/00317 , B01J2219/00479 , B01J2219/00481 , B01J2219/00484 , B01J2219/00495 , B01J2219/00585 , B01J2219/00599 , B01J2219/00691 , B01J2219/00743 , B01L3/5027 , B01L2200/027 , B01L2200/10 , B01L2300/0816 , B01L2300/0822 , B01L2300/0867 , B01L2300/10 , C12M23/16 , C12M23/58 , C12M29/04 , C12M29/10 , G01N21/31 , G01N21/6408 , G01N21/6428 , G01N21/6452 , G01N21/7703 , G01N35/0099 , G01N2021/6484 , G01N2021/7783 , G01N2021/7786
摘要： The present invention provides a variety of microscale bioreactors (microfermentors) and microscale bioreactor arrays for use in culturing cells. The microfermentors include a vessel for culturing cells and means for providing oxygen to the interior of the vessel at a concentration sufficient to support cell growth, e.g., growth of bacterial cells. Depending on the embodiment, the microfermentor vessel may have various interior volumes less than approximately 1 ml. The microfermentors may include an aeration membrane and optionally a variety of sensing devices. The invention further provides a chamber to contain the microfermentors and microfermentor arrays and to provide environmental control. Certain of the microfermentors include a second chamber that may be used, e.g., to provide oxygen, nutrients, pH control, etc., to the culture vessel and/or to remove metabolites, etc. Various methods of using the microfermentors, e.g., to select optimum cell strains or bioprocess parameters are provided.
摘要翻译：本发明提供用于培养细胞的各种微量生物反应器（微发酵器）和微生物生物反应器阵列。微发酵器包括用于培养细胞的容器和用于以足以支持细胞生长（例如细菌细胞生长）的浓度向容器内部提供氧气的装置。根据实施方案，微发酵罐容器可以具有小于约1ml的各种内部容积。微发酵器可以包括曝气膜和任选的各种感测装置。本发明进一步提供了容纳微发射体和微发光体阵列并提供环境控制的腔室。某些微发酵器包括第二室，其可以用于例如向培养容器提供氧气，营养物质，pH控制等，和/或去除代谢物等。使用微发酵器的各种方法，例如，选择最佳细胞株或生物过程参数。

5. 发明申请

WO2006037022A2 MICROBIOREACTOR FOR CONTINUOUS CELL CULTURE 审中-公开
标题翻译：连续细胞培养微生物
公开(公告)号：WO2006037022A2
公开(公告)日：2006-04-06
申请号：PCT/US2005/034774
申请日：2005-09-26
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , ZHANG, Zhiyu , BOCCAZZI, Paolo , CHOI, Hyun-Goo , JENSEN, Klavs, F. , SINSKEY, Anthony, J.
发明人： ZHANG, Zhiyu , BOCCAZZI, Paolo , CHOI, Hyun-Goo , JENSEN, Klavs, F. , SINSKEY, Anthony, J.
IPC分类号： C12M3/00
CPC分类号： C12M23/16 , B01L3/502707 , B01L3/502715 , B01L3/502723 , B01L2300/0654 , B01L2300/0816 , B01L2300/0877 , B01L2300/0887 , B01L2300/10 , B01L2300/185 , B01L2300/1883 , B01L2400/0466 , B01L2400/0487 , B82Y15/00 , B82Y30/00 , B82Y40/00 , C12M29/04 , C12M29/10 , C12M37/00 , C12M41/26 , C12M41/34 , G01N21/03 , G01N2021/0346
摘要： The present invention microscale bioreactors (microfermentors) and microscale bioreactor arrays for use in culturing cells. The microfermentors include a vessel for culturing cells and means for providing oxygen to the interior of the vessel at a concentration sufficient to support cell growth, e.g., growth of bacterial cells. Depending on the embodiment, the microfermentor vessel may have various interior volumes of less than approximately 1 ml. The microfermentors may include an aeration membrane and optionally a variety of sensing devices. Methods of using the microfermentors, e.g., to select optimum cell strains or bioprocess parameters are provided. The microbioreactors having a variety of different designs, some of which incorporate active fluid mixing and/or have the capability to operate in batch, fed-batch, or continuous mode. In certain embodiments the microreactors operate as microchemostats. Methods for culturing cells under chemostat conditions in a microbioreactor are also provided.
摘要翻译：本发明的用于培养细胞的微生物生物反应器（微发酵器）和微生物生物反应器阵列。微发酵器包括用于培养细胞的容器和用于以足以支持细胞生长（例如细菌细胞生长）的浓度向容器内部提供氧气的装置。取决于实施方案，微发酵罐容器可以具有小于约1ml的各种内部体积。微发酵器可以包括曝气膜和任选的各种感测装置。提供了使用微发酵器的方法，例如选择最佳细胞株或生物过程参数。具有各种不同设计的微生物反应器，其中一些结合有活性流体混合和/或具有以批量，补料分批或连续模式操作的能力。在某些实施方案中，微反应器作为微量化学剂操作。还提供了在微生物反应器中在恒化器条件下培养细胞的方法。

6. 发明申请

WO1988000948A1 METHOD TO CONTROL AND PRODUCE NOVEL BIOPOLYMERS 审中-公开
标题翻译：控制和生产新型生物聚合物的方法
公开(公告)号：WO1988000948A1
公开(公告)日：1988-02-11
申请号：PCT/US1987001835
申请日：1987-07-28
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY
发明人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , SINSKEY, Anthony, J. , EASSON, Donald, Davidson, Jr. , RHA, ChoKyun
IPC分类号： C07H15/12
CPC分类号： C09K8/905 , C12N15/52 , C12P19/04
摘要： A method for identifying, characterizing, utilizing and modifying a set of genes which interact to produce a specific polymer. The method is demonstrated for production of an exocellular polysaccharide produced by the bacteria Zoogloea ramigera. The isolated polysaccharide is useful as a viscosity modifier, oil field chemical, drag reducing agent, dispersant or flocculant. Modification of the isolated genes, for example, by insertion of a regulatable promoter, provides a means for alteration of the enzymes responsible for synthesis of the polysaccharide and its resulting structure.
摘要翻译：用于鉴定，表征，利用和修饰相互作用以产生特定聚合物的一组基因的方法。该方法被证明用于生产由细菌Zoogloea苎麻生产的外细胞多糖。分离的多糖可用作粘度调节剂，油田化学品，减阻剂，分散剂或絮凝剂。分离的基因的修饰，例如通过插入可调节的启动子，提供了改变负责合成多糖的酶及其所得结构的方法。

7. 发明申请

WO2007038572A2 MICROBIOREACTOR FOR CONTINUOUS CELL CULTURE 审中-公开
标题翻译：连续细胞培养微生物
公开(公告)号：WO2007038572A2
公开(公告)日：2007-04-05
申请号：PCT/US2006/037612
申请日：2006-09-26
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , ZHANG, Zhiyu , BOCCAZZI, Paolo , CHOI, Hyun-goo , JENSEN, Klavs, F. , SINSKEY, Anthony, J.
发明人： ZHANG, Zhiyu , BOCCAZZI, Paolo , CHOI, Hyun-goo , JENSEN, Klavs, F. , SINSKEY, Anthony, J.
IPC分类号： C12M3/00 , C12M1/12
CPC分类号： C12M23/16 , B01F13/0059 , B01F13/0827 , B01L3/502707 , B01L3/502715 , B01L3/502723 , B01L7/00 , B01L2300/0822 , B01L2300/0887 , B01L2300/10 , B82Y30/00 , C12M23/24 , C12M23/34 , C12M27/02 , C12M29/04 , C12M41/26 , C12M41/32
摘要： The present invention microscale bioreactors (microfermentors) and microscale bioreactor arrays for use in culturing cells. The microfermentors include a vessel for culturing cells and means for providing oxygen to the interior of the vessel at a concentration sufficient to support cell growth, e.g., growth of bacterial cells. Depending on the embodiment, the microfermentor vessel may have various interior volumes of less than approximately 1 ml. The microfermentors may include an aeration membrane and optionally a variety of sensing devices. Methods of using the microfermentors, e.g., to select optimum cell strains or bioprocess parameters are provided. The microbioreactors having a variety of different designs, some of which incorporate active fluid mixing and/or have the capability to operate in batch, fed- batch, or continuous mode. In certain embodiments the microreactors operate as microchemostats. Methods for culturing cells under chemostat conditions in a microbioreactor are also provided.
摘要翻译：本发明的用于培养细胞的微生物生物反应器（微发酵器）和微生物生物反应器阵列。微发酵器包括用于培养细胞的容器和用于以足以支持细胞生长（例如细菌细胞生长）的浓度向容器内部提供氧气的装置。取决于实施方案，微发酵罐容器可以具有小于约1ml的各种内部体积。微发酵器可以包括曝气膜和任选的各种感测装置。提供了使用微发酵器的方法，例如选择最佳细胞株或生物过程参数。具有各种不同设计的微生物反应器，其中一些包括主动流体混合和/或具有以批量，补料或连续模式操作的能力。在某些实施方案中，微反应器作为微量化学剂操作。还提供了在微生物反应器中在恒化器条件下培养细胞的方法。

8. 发明申请

WO1989009270A1 METHOD FOR ALTERING SURFACE CHARGE OF MICROORGANISMS 审中-公开
标题翻译：改善微生物表面电荷的方法
公开(公告)号：WO1989009270A1
公开(公告)日：1989-10-05
申请号：PCT/US1989001135
申请日：1989-03-20
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY
发明人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , EASSON, Donald, D., Jr. , SINSKEY, Anthony, J. , PEOPLES, Oliver, P.
IPC分类号： C12N15/00
CPC分类号： C12N9/00 , C12N9/10 , C12P19/00
摘要： The present invention is a method to produce polysaccharides with altered functional groups on their surface having modified surface charge as a result of the altered functional groups. These functional groups include, but are not limited to, negatively charged pyruvyl and succinyl moieties and positively charged amino groups. The polysaccharides can be produced by either: (1) mutation and selection of organisms synthesizing polysaccharides with modified surface charge or (2) engineering of specific genes within organisms to alter the charged groups attached to the polysaccharides during synthesis. Mutation of Zoogloea ramigera polysaccharide, as measured by the change in pyruvate content, is demonstrated. The isolation, characterization and manipulation of the gene for the pyruvyl transferase enzyme, which adds pyruvyl moities to the exopolysaccharide produced by Zoogloea ramigera, leading to an exopolysaccharide having a modified surface charge, is detailed. The modified exopolysaccharides have new applications due to the differences in surface charge. The modified enzymes and sequences regulating their expression can be used to produce novel polysaccharides. Further, studies of the structural-functional relationships of the modified exopolysaccharide can lead to a better understanding of secondary and tertiary structure in polysaccharides.
摘要翻译：本发明是由于功能基团的改变而产生具有改变的表面电荷的表面上具有改变的官能团的多糖的方法。这些官能团包括但不限于带负电荷的丙烯酰基和琥珀酰基部分和带正电荷的氨基。多糖可以通过：（1）突变和选择具有修饰的表面电荷合成多糖的生物体，或（2）在生物体内工程化特定基因以改变在合成过程中与多糖连接的带电基团。证明了通过丙酮酸含量变化测量的Zoogloea苎麻多糖的突变。详细说明了分离，表征和操作丙酮酸转移酶的基因，其向由苎麻生产的外多糖添加丙酮酸，导致具有改性表面电荷的外多糖。由于表面电荷的差异，修饰的外多糖具有新的应用。修饰的酶和调节其表达的序列可用于产生新型多糖。此外，修饰的外多糖的结构 - 功能关系的研究可以更好地了解多糖中的二级和三级结构。

9. 发明申请

WO1993009225A1 METHOD AND DEREGULATED ENZYME FOR THREONINE PRODUCTION 审中-公开
标题翻译：用于生产THR生产的方法和分离酶
公开(公告)号：WO1993009225A1
公开(公告)日：1993-05-13
申请号：PCT/US1992009325
申请日：1992-10-30
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY
发明人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , ARCHER, John, A., C. , FOLLETTIE, Maximillian, T. , SINSKEY, Anthony, J.
IPC分类号： C12N09/04
CPC分类号： C12N9/0006 , C12N9/12 , C12N9/88 , C12N15/52 , C12N15/77 , C12P13/08
摘要： Mutagenesis of the gene encoding homoserine dehydrogenase (hom) for production of the amino acid threonine is described. The mutation causes an alteration in the carboxy terminus of the enzyme that interferes with end-product inhibition by threonine. The lack of end-product inhibition causes an overproduction of threonine.
摘要翻译：描述了用于产生氨基酸苏氨酸的编码高丝氨酸脱氢酶（hom）的基因的突变。该突变导致酶的羧基末端的改变，其干扰苏氨酸的最终产物抑制。最终产物抑制的缺乏导致苏氨酸的过量产生。

10. 发明申请

WO1989000202A2 METHOD FOR PRODUCING NOVEL POLYESTER BIOPOLYMERS 审中-公开
标题翻译：生产新型聚酯生物聚合物的方法
公开(公告)号：WO1989000202A2
公开(公告)日：1989-01-12
申请号：PCT/US1988002227
申请日：1988-06-27
申请人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY
发明人： MASSACHUSETTS INSTITUTE OF TECHNOLOGY , PEOPLES, Oliver, P. , SINSKEY, Anthony, J.
IPC分类号： C12P07/62
CPC分类号： C12N9/0006 , C12N9/00 , C12N9/1029 , C12N15/52 , C12P7/625
摘要： The present invention is a method for controlling biopolymer synthesis by determining the genetics and enzymology of polyhydroxybutyrate (PHB) biosynthesis at the molecular level. The purified enzymes and genes provide the means for developing new PHB-like biopolymers having polyester backbones. Specific aims are to 1) control the chain length of the polymers produced in fermentation processes through genetic manipulation, 2) incorporate different monomers into the polymers to produce co-polymers with different physical properties, and 3) examine the physical/rheological properties of these new biopolymers in order to develop further design criteria at the molecular level. The method for engineering biopolymer synthesis includes: isolation and characterization of the genes for the enzymes in the synthetic pathway (beta-ketothiolase, acetoacetyl-CoA reductase and PHB synthetase); cloning of the genes in a vector(s); placement of the vector(s) under the control of regulated promoters; expression of the genes; determination of the function and use of other factors such as substrate specificity in polymer production and composition; and isolation and physical and chemical analysis of the resulting polymers.
摘要翻译：本发明是通过在分子水平上测定聚羟基丁酸（PHB）生物合成的遗传学和酶学方法来控制生物聚合物合成的方法。纯化的酶和基因为开发具有聚酯主链的新型PHB样生物聚合物提供了手段。具体目的是1）通过遗传操作控制在发酵过程中产生的聚合物的链长，2）将不同单体引入聚合物中以产生具有不同物理性质的共聚物，和3）检查这些物质/流变学性质新的生物聚合物，以便在分子水平上制定进一步的设计标准。工程生物聚合物合成的方法包括：合成途径中酶的分离和表征（β-酮硫解酶，乙酰乙酰辅酶A还原酶和PHB合成酶）; 在载体中克隆基因; 将载体置于受调节启动子的控制下; 基因表达; 确定聚合物生产和组成中其他因素如底物特异性的功能和用途; 以及所得聚合物的分离和物理和化学分析。

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