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1. 发明授权

US08715988B2 Alkane oxidation by modified hydroxylases 有权
标题翻译：改性羟化酶的烷烃氧化
公开(公告)号：US08715988B2
公开(公告)日：2014-05-06
申请号：US11697404
申请日：2007-04-06
申请人： Frances Arnold , Peter Meinhold , Matthew W. Peters , Rudi Fasan , Mike M. Y. Chen
发明人： Frances Arnold , Peter Meinhold , Matthew W. Peters , Rudi Fasan , Mike M. Y. Chen
IPC分类号： C12N9/02
CPC分类号： C12P7/04 , C07K14/32 , C12N9/0042 , C12N9/0071 , C12N9/0077 , C12P7/06 , C12Y106/02004 , C12Y114/14001 , C12Y114/15 , C12Y114/15003 , Y02P20/52
摘要： This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.
摘要翻译：本发明涉及改性羟化酶。本发明还涉及表达这种修饰的羟化酶的细胞和通过使合适的底物与这种细胞接触来制备羟基化烷烃的方法。

2. 发明申请

US20080057577A1 Alkane Oxidation By Modified Hydroxylases 有权
标题翻译：通过改性羟基酶进行的烷基氧化
公开(公告)号：US20080057577A1
公开(公告)日：2008-03-06
申请号：US11697404
申请日：2007-04-06
申请人： Frances Arnold , Peter Meinhold , Matthew Peters , Rudi Fasan , Mike Chen
发明人： Frances Arnold , Peter Meinhold , Matthew Peters , Rudi Fasan , Mike Chen
IPC分类号： C07K14/32 , C07K1/107 , C12N1/19 , C12N1/21 , C12N5/10
CPC分类号： C12P7/04 , C07K14/32 , C12N9/0042 , C12N9/0071 , C12N9/0077 , C12P7/06 , C12Y106/02004 , C12Y114/14001 , C12Y114/15 , C12Y114/15003 , Y02P20/52
摘要： This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.
摘要翻译：本发明涉及改性羟化酶。本发明还涉及表达这种修饰的羟化酶的细胞和通过使合适的底物与这种细胞接触来制备羟基化烷烃的方法。

3. 发明申请

US20100143997A1 ENGINEERED MICROORGANISMS CAPABLE OF PRODUCING TARGET COMPOUNDS UNDER ANAEROBIC CONDITIONS 审中-公开
标题翻译：在无机条件下生产目标化合物的工程微生物
公开(公告)号：US20100143997A1
公开(公告)日：2010-06-10
申请号：US12610784
申请日：2009-11-02
申请人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Eva Eckl , Andrew Hawkins , Aristos Aristidou , Catherine Asleson Dundon , Doug Lies , Sabine Bastian , Frances Arnold , Jun Urano
发明人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Eva Eckl , Andrew Hawkins , Aristos Aristidou , Catherine Asleson Dundon , Doug Lies , Sabine Bastian , Frances Arnold , Jun Urano
IPC分类号： C12P7/16 , C12N9/02 , C12N1/21 , C12N1/15 , C12N1/19
CPC分类号： C12P7/16 , C12N9/0006 , C12N9/0008 , C12N9/0036 , Y02E50/10 , Y02E50/17
摘要： The present invention is generally provides recombinant microorganisms comprising engineered metabolic pathways capable of producing C3-C5 alcohols under aerobic and anaerobic conditions. The invention further provides ketol-acid reductoisomerase enzymes which have been mutated or modified to increase their NADH-dependent activity or to switch the cofactor preference from NADPH to NADH and are expressed in the modified microorganisms. In addition, the invention provides isobutyraldehyde dehydrogenase enzymes expressed in modified microorganisms. Also provided are methods of producing beneficial metabolites under aerobic and anaerobic conditions by contacting a suitable substrate with the modified microorganisms of the present invention.
摘要翻译：本发明通常提供重组微生物，其包含能够在需氧和厌氧条件下生产C3-C5醇的工程化代谢途径。本发明还提供已被突变或修饰以提高其NADH依赖性活性或将辅因子偏好从NADPH转变为NADH并在修饰的微生物中表达的酮酸酸还原异构酶。此外，本发明提供了以修饰微生物表达的异丁醛脱氢酶。还提供了在需氧和厌氧条件下通过使合适的底物与本发明的改性微生物接触来生产有益代谢物的方法。

4. 发明申请

US20120190089A1 ENGINEERED MICROOGRANISMS CAPABLE OF PRODUCING TARGET COMPOUNDS UNDER ANAEROBIC CONDITIONS 审中-公开
标题翻译：在无机条件下生产目标化合物的工程化微生物
公开(公告)号：US20120190089A1
公开(公告)日：2012-07-26
申请号：US13303884
申请日：2011-11-23
申请人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew C. Hawkins , Sabine Bastian , Frances Arnold , Jun Urano
发明人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew C. Hawkins , Sabine Bastian , Frances Arnold , Jun Urano
IPC分类号： C12P7/16 , C12N1/19
CPC分类号： C12P7/16 , C12N9/0006 , C12N9/0008 , C12N9/0036 , Y02E50/10
摘要： The present invention is generally provides recombinant microorganisms comprising engineered metabolic pathways capable of producing C3-C5 alcohols under aerobic and anaerobic conditions. The invention further provides ketol-acid reductoisomerase enzymes which have been mutated or modified to increase their NADH-dependent activity or to switch the cofactor preference from NADPH to NADH and are expressed in the modified microorganisms. In addition, the invention provides isobutyraldehyde dehydrogenase enzymes expressed in modified microorganisms. Also provided are methods of producing beneficial metabolites under aerobic and anaerobic conditions by contacting a suitable substrate with the modified microorganisms of the present invention.
摘要翻译：本发明通常提供重组微生物，其包含能够在需氧和厌氧条件下生产C3-C5醇的工程化代谢途径。本发明还提供已被突变或修饰以提高其NADH依赖性活性或将辅因子偏好从NADPH转变为NADH并在修饰的微生物中表达的酮酸酸还原异构酶。此外，本发明提供以修饰微生物表达的异丁醛脱氢酶。还提供了在需氧和厌氧条件下通过使合适的底物与本发明的改性微生物接触来生产有益代谢物的方法。

5. 发明授权

US08097440B1 Engineered microorganisms capable of producing target compounds under anaerobic conditions 有权
标题翻译：能够在厌氧条件下产生目标化合物的工程微生物
公开(公告)号：US08097440B1
公开(公告)日：2012-01-17
申请号：US13269937
申请日：2011-10-10
申请人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew C. Hawkins , Jun Urano , Sabine Bastian , Frances Arnold
发明人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew C. Hawkins , Jun Urano , Sabine Bastian , Frances Arnold
IPC分类号： C12P7/16 , C12N15/53 , C12N1/15 , C12N9/02
CPC分类号： C12P7/16 , C12N9/0006 , C12N9/0008 , C12N9/0036 , Y02E50/10 , Y02E50/17
摘要： The present invention is generally provides recombinant microorganisms comprising engineered metabolic pathways capable of producing C3-C5 alcohols under aerobic and anaerobic conditions. The invention further provides ketol-acid reductoisomerase enzymes which have been mutated or modified to increase their NADH-dependent activity or to switch the cofactor preference from NADPH to NADH and are expressed in the modified microorganisms. In addition, the invention provides isobutyraldehyde dehydrogenase enzymes expressed in modified microorganisms. Also provided are methods of producing beneficial metabolites under aerobic and anaerobic conditions by contacting a suitable substrate with the modified microorganisms of the present invention.
摘要翻译：本发明通常提供重组微生物，其包含能够在需氧和厌氧条件下生产C3-C5醇的工程化代谢途径。本发明还提供已被突变或修饰以提高其NADH依赖性活性或将辅因子偏好从NADPH转变为NADH并在修饰的微生物中表达的酮酸酸还原异构酶。此外，本发明提供以修饰微生物表达的异丁醛脱氢酶。还提供了在需氧和厌氧条件下通过使合适的底物与本发明的改性微生物接触来生产有益代谢物的方法。

6. 发明申请

US20050059128A1 Regio- and enantioselective alkane hydroxylation with modified cytochrome P450 有权
标题翻译：修饰的细胞色素P450的区域和对映选择性烷烃羟基化
公开(公告)号：US20050059128A1
公开(公告)日：2005-03-17
申请号：US10869825
申请日：2004-06-15
申请人： Frances Arnold , Matthew Peters , Peter Meinhold
发明人： Frances Arnold , Matthew Peters , Peter Meinhold
IPC分类号： C07H21/04 , C12N20060101 , C12N9/02
CPC分类号： C12N9/0071 , C12N9/0077 , C12P7/04 , C12Y114/14001
摘要： Cytochrome P450 BM-3 from Bacillus megaterium was engineered using a combination of directed evolution and site-directed mutagenesis to hydroxylate linear alkanes regio- and enantioselectively using atmospheric dioxygen as an oxidant. Mutant 9-10A-A328V hydroxylates octane primarily at the 2-positio to form S-2-octanol (40% ee). Another mutant, 1-12G, hydroxylates alkanes larger than hexane primarily at the 2-position, but forms R-2-alcohols (40-55% ee). These biocatalysts are highly active for alkane substrates and support thousands of product turnovers. These regio- and enantio-selectivities are retained in whole-cell biotransformations with E. coli, where the engineered P450s can be expressed at high levels and the expensive cofactor is supplied endogenously.
摘要翻译：使用定向进化和定点诱变的组合将来自巨大芽孢杆菌的细胞色素P450 BM-3设计为使用大气二氧氧化物作为区域和对映选择性的羟基化线型烷烃。突变体9-10A-A328V主要在2-位羟基化辛烷值以形成S-2-辛醇（40％ee）。另一个突变体1-12G，主要在2-位羟基化烷烃大于己烷，但形成R-2-醇（40-55％ee）。这些生物催化剂对烷烃底物具有高活性，并支持数千种产品周转。这些区域和对映体选择性保留在与大肠杆菌的全细胞生物转化中，其中工程化的P450可以以高水平表达并且昂贵的辅因子是内源性提供的。

7. 发明申请

US20120058532A1 ENGINEERED MICROORGANISMS CAPABLE OF PRODUCING TARGET COMPOUNDS UNDER ANAEROBIC CONDITIONS 审中-公开
标题翻译：在无机条件下生产目标化合物的工程微生物
公开(公告)号：US20120058532A1
公开(公告)日：2012-03-08
申请号：US13271084
申请日：2011-10-11
申请人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew Hawkins , Sabine Bastian , Frances Arnold , Jun Urano
发明人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew Hawkins , Sabine Bastian , Frances Arnold , Jun Urano
IPC分类号： C12P7/16 , C12N1/19 , C12N15/53
CPC分类号： C12P7/16 , C12N9/0006 , C12N9/0008 , C12N9/0036 , Y02E50/10 , Y02E50/17
摘要： The present invention is generally provides recombinant microorganisms comprising engineered metabolic pathways capable of producing C3-C5 alcohols under aerobic and anaerobic conditions. The invention further provides ketol-acid reductoisomerase enzymes which have been mutated or modified to increase their NADH-dependent activity or to switch the cofactor preference from NADPH to NADH and are expressed in the modified microorganisms. In addition, the invention provides isobutyraldehyde dehydrogenase enzymes expressed in modified microorganisms. Also provided are methods of producing beneficial metabolites under aerobic and anaerobic conditions by contacting a suitable substrate with the modified microorganisms of the present invention.
摘要翻译：本发明通常提供重组微生物，其包含能够在需氧和厌氧条件下生产C3-C5醇的工程化代谢途径。本发明还提供已被突变或修饰以提高其NADH依赖性活性或将辅因子偏好从NADPH转变为NADH并在修饰的微生物中表达的酮酸酸还原异构酶。此外，本发明提供了以修饰微生物表达的异丁醛脱氢酶。还提供了在需氧和厌氧条件下通过使合适的底物与本发明的改性微生物接触来生产有益代谢物的方法。

8. 发明授权

US09012189B2 Modified alcohol dehydrogenases for the production of fuels and chemicals 有权
标题翻译：用于生产燃料和化学品的改性醇脱氢酶
公开(公告)号：US09012189B2
公开(公告)日：2015-04-21
申请号：US13025805
申请日：2011-02-11
申请人： Sabine Bastian , Frances Arnold , Peter Meinhold
发明人： Sabine Bastian , Frances Arnold , Peter Meinhold
IPC分类号： C12P7/16 , C12N15/81 , C12N9/04
CPC分类号： C12N15/81 , C12N9/0006 , C12P7/16 , C12Y101/01 , C12Y102/01005 , Y02E50/10 , Y02P20/52
摘要： The present invention relates to recombinant microorganisms comprising biosynthetic pathways and methods of using said recombinant microorganisms to produce various beneficial metabolites. In various aspects of the invention, the recombinant microorganisms may further comprise one or more modifications resulting in the reduction or elimination of 3 keto-acid (e.g., acetolactate and 2-aceto-2-hydroxybutyrate) and/or aldehyde-derived by-products. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.
摘要翻译：本发明涉及包含生物合成途径的重组微生物和使用所述重组微生物产生各种有益代谢物的方法。在本发明的各个方面，重组微生物可以进一步包含一种或多种修饰，导致还原或消除3个酮酸（例如乙酰乳酸和2-乙酰-2-羟基丁酸酯）和/或醛衍生的副产物。在本文所述的各种实施方案中，重组微生物可以是酵母进化枝，Crabtree阴性酵母微生物，Crabtree阳性酵母微生物，WGD（全基因组复制）酵母微生物，WGD前（全基因组复制）酵母微生物，和非发酵酵母微生物。

9. 发明申请

US20140017748A1 MODIFIED ALCOHOL DEHYDROGENASES FOR THE PRODUCTION OF FUELS AND CHEMICALS 审中-公开
标题翻译：用于生产燃料和化学品的改性酒精脱氢剂
公开(公告)号：US20140017748A1
公开(公告)日：2014-01-16
申请号：US13431532
申请日：2012-03-27
申请人： Sabine BASTIAN , Frances Arnold , Peter Meinhold
发明人： Sabine BASTIAN , Frances Arnold , Peter Meinhold
IPC分类号： C12P7/16
CPC分类号： C12P7/16 , C12N9/0006 , Y02E50/10
摘要： The present invention relates to recombinant microorganisms comprising biosynthetic pathways and methods of using said recombinant microorganisms to produce various beneficial metabolites. In various aspects of the invention, the recombinant microorganisms may further comprise one or more modifications resulting in the reduction or elimination of 3 keto-acid (e.g., acetolactate and 2-aceto-2-hydroxybutyrate) and/or aldehyde-derived by-products. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.
摘要翻译：本发明涉及包含生物合成途径的重组微生物和使用所述重组微生物产生各种有益代谢物的方法。在本发明的各个方面，重组微生物可以进一步包含一种或多种修饰，导致还原或消除3个酮酸（例如乙酰乳酸和2-乙酰-2-羟基丁酸酯）和/或醛衍生的副产物。在本文所述的各种实施方案中，重组微生物可以是酵母进化枝，Crabtree阴性酵母微生物，Crabtree阳性酵母微生物，WGD（全基因组复制）酵母微生物，WGD前（全基因组复制）酵母微生物，和非发酵酵母微生物。

10. 发明申请

US20120028324A1 ENGINEERED MICROORGANISMS CAPABLE OF PRODUCING TARGET COMPOUNDS UNDER ANAEROBIC CONDITIONS 有权
标题翻译：在无机条件下生产目标化合物的工程微生物
公开(公告)号：US20120028324A1
公开(公告)日：2012-02-02
申请号：US13269937
申请日：2011-10-10
申请人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew Hawkins , Sabine Bastian , Frances Arnold , Jun Urano
发明人： Thomas Buelter , Peter Meinhold , Reid M. Renny Feldman , Andrew Hawkins , Sabine Bastian , Frances Arnold , Jun Urano
IPC分类号： C12P7/16 , C12N1/00 , C12N1/19 , C07H21/04
CPC分类号： C12P7/16 , C12N9/0006 , C12N9/0008 , C12N9/0036 , Y02E50/10 , Y02E50/17
摘要： The present invention is generlly provides recombinant microorganisms comprising engineered metabolic pathways capable of producing C3-C5 alcohols under aerobic and anaerobic conditions. The invention further provides ketol-acid reductoisomerase enzymes which have been mutated or modified to increase their NADH-dependent activity or to switch the cofactor preference from NADPH to NADH and are expressed in the modified microorganisms. In addition, the invention provides isobutyraldehyde dehydrogenase enzymes expressed in modified microorganisms. Also provided are methods of producing beneficial metabolites under aerobic and anaerobic conditions by contacting a suitable substrate with the modified microorganisms of the present invention.
摘要翻译：本发明总体上提供了重组微生物，其包含能够在有氧和厌氧条件下产生C3-C5醇的工程化代谢途径。本发明还提供已被突变或修饰以提高其NADH依赖性活性或将辅因子偏好从NADPH转变为NADH并在修饰的微生物中表达的酮酸酸还原异构酶。此外，本发明提供了以修饰微生物表达的异丁醛脱氢酶。还提供了在需氧和厌氧条件下通过使合适的底物与本发明的改性微生物接触来生产有益代谢物的方法。

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