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公开(公告)号：US08603800B2
公开(公告)日：2013-12-10
申请号：US13752293
申请日：2013-01-28
Applicant: Amyris, Inc.
Inventor： Timothy Stevens Gardner , Kristy Michelle Hawkins , Adam Leon Meadows , Annie Ening Tsong , Yoseph Tsegaye
CPC classification number: C12P5/007 , C12N9/0006 , C12N9/0008 , C12N9/1288 , C12N15/52 , C12N15/81 , C12P7/42 , C12P23/00 , C12Y101/01034 , C12Y102/0101 , C12Y207/08
Abstract: Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.
Abstract translation: 本文提供了在宿主细胞中异源产生乙酰辅酶A衍生的类异戊二烯的组合物和方法。 在一些实施方案中，宿主细胞被遗传修饰以包含编码乙醛脱氢酶，乙酰化（ADA，E.C.1.2.1.10）和包含使用NADH的HMG-CoA还原酶的MEV途径的异源核苷酸序列。 在一些实施方案中，宿主细胞被遗传修饰以包含编码包含乙酰乙酰辅酶A合成酶的ADA和MEV途径的异源核苷酸序列。 在一些实施方案中，经遗传修饰的宿主细胞还包含一个或多个编码磷酸酮醇酶和磷酸转乙酰酶的异源核苷酸序列。 在一些实施方案中，遗传修饰的宿主细胞还包括天然PDH旁路的功能性破坏。 本文描述的组合物和方法提供了用于异源产生乙酰辅酶A衍生的类异戊二烯的能量效率尚未的氧化还原平衡途径。
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公开(公告)号：US09914941B2
公开(公告)日：2018-03-13
申请号：US14474976
申请日：2014-09-02
Applicant: Amyris, Inc.
Inventor： Timothy Stevens Gardner , Kristy Michelle Hawkins , Adam Leon Meadows , Annie Ening Tsong , Yoseph Tsegaye
CPC classification number: C12P5/007 , C12N9/0006 , C12N9/0008 , C12N9/1288 , C12N15/52 , C12N15/81 , C12P7/42 , C12P23/00 , C12Y101/01034 , C12Y102/0101 , C12Y207/08
Abstract: Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.
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公开(公告)号：US20140154765A1
公开(公告)日：2014-06-05
申请号：US14062798
申请日：2013-10-24
Applicant: AMYRIS, INC.
Inventor： Timothy Stevens GARDNER , Kristy Michelle Hawkins , Adam Leon Meadows , Annie Ening Tsong , Yoseph Tsegaye
CPC classification number: C12P5/007 , C12N9/0006 , C12N9/0008 , C12N9/1288 , C12N15/52 , C12N15/81 , C12P7/42 , C12P23/00 , C12Y101/01034 , C12Y102/0101 , C12Y207/08
Abstract: Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.
Abstract translation: 本文提供了在宿主细胞中异源产生乙酰辅酶A衍生的类异戊二烯的组合物和方法。 在一些实施方案中，宿主细胞被遗传修饰以包含编码乙醛脱氢酶，乙酰化（ADA，E.C.1.2.1.10）和包含使用NADH的HMG-CoA还原酶的MEV途径的异源核苷酸序列。 在一些实施方案中，宿主细胞被遗传修饰以包含编码包含乙酰乙酰辅酶A合成酶的ADA和MEV途径的异源核苷酸序列。 在一些实施方案中，经遗传修饰的宿主细胞还包含一个或多个编码磷酸酮醇酶和磷酸转乙酰酶的异源核苷酸序列。 在一些实施方案中，遗传修饰的宿主细胞还包括天然PDH旁路的功能性破坏。 本文描述的组合物和方法提供了用于异源产生乙酰辅酶A衍生的类异戊二烯的能量效率尚未的氧化还原平衡途径。
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公开(公告)号：US20190017074A1
公开(公告)日：2019-01-17
申请号：US16044381
申请日：2018-07-24
Applicant: Amyris, Inc.
Inventor： Andrew Horwitz , Kristy Michelle Hawkins , Max Schubert , Wayne Szeto
IPC: C12N15/90 , C12Q1/6897 , C12N15/11
Abstract: Provided herein are methods of integrating one or more exogenous nucleic acids into one or more selected target sites of a host cell genome. In certain embodiments, the methods comprise contacting the host cell genome with one or more integration polynucleotides comprising an exogenous nucleic acid to be integrated into a genomic target site, a nuclease capable of causing a break at the genomic target site, and a linear nucleic acid capable of homologous recombination with itself or with one or more additional linear nucleic acids contacted with the population of cells, whereupon said homologous recombination results in formation of a circular extrachromosomal nucleic acid comprising a coding sequence for a selectable marker. In some embodiments, the methods further comprise selecting a host cell that expresses the selectable marker.
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公开(公告)号：US20150184199A1
公开(公告)日：2015-07-02
申请号：US14577997
申请日：2014-12-19
Applicant: Amyris, Inc.
Inventor： Andrew Horwitz , Kristy Michelle Hawkins , Max Schubert , Wayne Szeto
CPC classification number: C12N15/907 , C12N15/111 , C12N15/902 , C12N2310/20 , C12Q1/6897
Abstract: Provided herein are methods of integrating one or more exogenous nucleic acids into one or more selected target sites of a host cell genome. In certain embodiments, the methods comprise contacting the host cell genome with one or more integration polynucleotides comprising an exogenous nucleic acid to be integrated into a genomic target site, a nuclease capable of causing a break at the genomic target site, and a linear nucleic acid capable of homologous recombination with itself or with one or more additional linear nucleic acids contacted with the population of cells, whereupon said homologous recombination results in formation of a circular extrachromosomal nucleic acid comprising a coding sequence for a selectable marker. In some embodiments, the methods further comprise selecting a host cell that expresses the selectable marker.
Abstract translation: 本文提供将一种或多种外源核酸整合到宿主细胞基因组的一个或多个选定靶位点的方法。 在某些实施方案中，所述方法包括使宿主细胞基因组与包含待整合到基因组靶位点的外源核酸的一个或多个整合多核苷酸，能够在基因组靶位点引起断裂的核酸酶和线性核酸 能够与其自身或与一个或多个与细胞群接触的另外的线性核酸进行同源重组，于是所述同源重组导致形成包含可选择标记的编码序列的环状染色体外核酸。 在一些实施方案中，所述方法还包括选择表达可选择标记的宿主细胞。
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Patent Agency Ranking
公开(公告)号：US10041092B2
公开(公告)日：2018-08-07
申请号：US15261727
申请日：2016-09-09
Applicant: Amyris, Inc.
Inventor： Andrew Horwitz , Kristy Michelle Hawkins , Max Schubert , Wayne Szeto
IPC: C12N9/22 , C12N15/90 , C12Q1/6897 , C12N15/11
Abstract: Provided herein are methods of integrating one or more exogenous nucleic acids into one or more selected target sites of a host cell genome. In certain embodiments, the methods comprise contacting the host cell genome with one or more integration polynucleotides comprising an exogenous nucleic acid to be integrated into a genomic target site, a nuclease capable of causing a break at the genomic target site, and a linear nucleic acid capable of homologous recombination with itself or with one or more additional linear nucleic acids contacted with the population of cells, whereupon said homologous recombination results in formation of a circular extrachromosomal nucleic acid comprising a coding sequence for a selectable marker. In some embodiments, the methods further comprise selecting a host cell that expresses the selectable marker.
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7.发明授权Use of phosphoketolase and phosphotransacetylase for production of acetyl-coenzyme A derived compounds 有权使用磷酸酮醇酶和磷酸转乙酰酶生产乙酰辅酶A衍生化合物公开(公告)号：US09410214B2
公开(公告)日：2016-08-09
申请号：US14214062
申请日：2014-03-14
Applicant: Amyris, Inc.
Inventor： Kristy Michelle Hawkins , Tina Tipawan Mahatdejkul-Meadows , Adam Leon Meadows , Lauren Barbara Pickens , Anna Tai , Annie Ening Tsong
CPC classification number: C12Y401/02009 , C12N9/1029 , C12N9/16 , C12N9/88 , C12P5/007 , C12P7/54 , C12Y203/01008 , C12Y301/03021 , Y02P20/52
Abstract: Provided herein are compositions and methods for improved production of acetyl-CoA and acetyl-CoA derived compounds in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding a phosphoketolase (PK), and a functional disruption of an endogenous enzyme that converts acetyl phosphate to acetate. In some embodiments, the host cell further comprises a heterologous nucleotide sequence encoding a phosphotransacetylase (PTA). In some embodiments, the enzyme that converts acetyl phosphate to acetate is a glycerol-1-phosphatase. In some embodiments, the glycerol-1-phosphatase is GPP1/RHR2. In some embodiments, the glycerol-1-phosphatase is GPP2/HOR2. The compositions and methods described herein provide an efficient route for the heterologous production of acetyl-CoA-derived compounds, including but not limited to, isoprenoids, polyketides, and fatty acids.
Abstract translation: 本文提供了用于在宿主细胞中改进乙酰辅酶A和乙酰辅酶A衍生化合物生产的组合物和方法。 在一些实施方案中，宿主细胞经遗传修饰以包含编码磷酸酮糖醇酶（PK）的异源核苷酸序列，以及将乙酰磷酸酯转化为乙酸盐的内源酶的功能性破坏。 在一些实施方案中，宿主细胞还包含编码磷酸转乙酰酶（PTA）的异源核苷酸序列。 在一些实施方案中，将乙酰磷酸酯转化为乙酸酯的酶是甘油-1-磷酸酶。 在一些实施方案中，甘油-1-磷酸酶是GPP1 / RHR2。 在一些实施方案中，甘油-1-磷酸酶是GPP2 / HOR2。 本文描述的组合物和方法为异源生产乙酰辅酶A衍生化合物（包括但不限于类异戊二烯，聚酮化合物和脂肪酸）提供了有效途径。
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公开(公告)号：US08415136B1
公开(公告)日：2013-04-09
申请号：US13673819
申请日：2012-11-09
Applicant: Amyris, Inc.
Inventor： Timothy Stevens Gardner , Kristy Michelle Hawkins , Adam Leon Meadows , Annie Ening Tsong , Yoseph Tsegaye
CPC classification number: C12P5/007 , C12N9/0006 , C12N9/0008 , C12N9/1288 , C12N15/52 , C12N15/81 , C12P7/42 , C12P23/00 , C12Y101/01034 , C12Y102/0101 , C12Y207/08
Abstract: Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.
Abstract translation: 本文提供了在宿主细胞中异源产生乙酰辅酶A衍生的类异戊二烯的组合物和方法。 在一些实施方案中，宿主细胞被遗传修饰以包含编码乙醛脱氢酶，乙酰化（ADA，E.C.1.2.1.10）和包含使用NADH的HMG-CoA还原酶的MEV途径的异源核苷酸序列。 在一些实施方案中，宿主细胞被遗传修饰以包含编码包含乙酰乙酰辅酶A合成酶的ADA和MEV途径的异源核苷酸序列。 在一些实施方案中，经遗传修饰的宿主细胞还包含一个或多个编码磷酸酮醇酶和磷酸转乙酰酶的异源核苷酸序列。 在一些实施方案中，遗传修饰的宿主细胞还包括天然PDH旁路的功能性破坏。 本文描述的组合物和方法提供了用于异源产生乙酰辅酶A衍生的类异戊二烯的能量效率尚未的氧化还原平衡途径。
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公开(公告)号：US20170058299A1
公开(公告)日：2017-03-02
申请号：US15261727
申请日：2016-09-09
Applicant: Amyris, Inc.
Inventor： Andrew Horwitz , Kristy Michelle Hawkins , Max Schubert , Wayne Szeto
IPC: C12N15/90
CPC classification number: C12N15/907 , C12N15/111 , C12N15/902 , C12N2310/20 , C12Q1/6897
Abstract: Provided herein are methods of integrating one or more exogenous nucleic acids into one or more selected target sites of a host cell genome. In certain embodiments, the methods comprise contacting the host cell genome with one or more integration polynucleotides comprising an exogenous nucleic acid to be integrated into a genomic target site, a nuclease capable of causing a break at the genomic target site, and a linear nucleic acid capable of homologous recombination with itself or with one or more additional linear nucleic acids contacted with the population of cells, whereupon said homologous recombination results in formation of a circular extrachromosomal nucleic acid comprising a coding sequence for a selectable marker. In some embodiments, the methods further comprise selecting a host cell that expresses the selectable marker.
Abstract translation: 本文提供将一种或多种外源核酸整合到宿主细胞基因组的一个或多个选定靶位点的方法。 在某些实施方案中，所述方法包括使宿主细胞基因组与包含待整合到基因组靶位点的外源核酸的一个或多个整合多核苷酸，能够在基因组靶位点引起断裂的核酸酶和线性核酸 能够与其自身或与一个或多个与细胞群接触的另外的线性核酸进行同源重组，于是所述同源重组导致形成包含可选择标记的编码序列的环状染色体外核酸。 在一些实施方案中，所述方法还包括选择表达可选择标记的宿主细胞。
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公开(公告)号：US09476065B2
公开(公告)日：2016-10-25
申请号：US14577997
申请日：2014-12-19
Applicant: Amyris, Inc.
Inventor： Andrew Horwitz , Kristy Michelle Hawkins , Max Schubert , Wayne Szeto
CPC classification number: C12N15/907 , C12N15/111 , C12N15/902 , C12N2310/20 , C12Q1/6897
Abstract: Provided herein are methods of integrating one or more exogenous nucleic acids into one or more selected target sites of a host cell genome. In certain embodiments, the methods comprise contacting the host cell genome with one or more integration polynucleotides comprising an exogenous nucleic acid to be integrated into a genomic target site, a nuclease capable of causing a break at the genomic target site, and a linear nucleic acid capable of homologous recombination with itself or with one or more additional linear nucleic acids contacted with the population of cells, whereupon said homologous recombination results in formation of a circular extrachromosomal nucleic acid comprising a coding sequence for a selectable marker. In some embodiments, the methods further comprise selecting a host cell that expresses the selectable marker.
Abstract translation: 本文提供将一种或多种外源核酸整合到宿主细胞基因组的一个或多个选定靶位点的方法。 在某些实施方案中，所述方法包括使宿主细胞基因组与包含待整合到基因组靶位点的外源核酸的一个或多个整合多核苷酸，能够在基因组靶位点引起断裂的核酸酶和线性核酸 能够与其自身或与一个或多个与细胞群接触的另外的线性核酸进行同源重组，于是所述同源重组导致形成包含可选择标记的编码序列的环状染色体外核酸。 在一些实施方案中，所述方法还包括选择表达可选择标记的宿主细胞。
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