公开(公告)号：US11685908B2

公开(公告)日：2023-06-27

申请号：US16978629

申请日：2019-03-08

Applicant: Genomatica, Inc.

Inventor： Michael A. Noble ,  Kevin G. Hoff ,  Anna Lechner ,  Harish Nagarajan

IPC: C12N9/10 ,  C12P7/22 ,  C12P7/42

CPC classification number: C12N9/1085 ,  C12P7/22 ,  C12P7/42

Abstract: Described herein are prenyltransferases including non-natural variants thereof having at least one amino acid substitution as compared to its corresponding natural or unmodified prenyltransferases and that are capable of at least two-fold greater rate of formation of cannabinoids such as cannabigerolic acid, cannabigerovarinic acid, cannabigerorcinic acid, and cannabigerol, as compared to a wild type control. Prenyltransferase variants also demonstrated regioselectivity to desired cannabinoid isomers such as CDBA (3-GOLA), 3-GDVA, 3-GOSA, and CBG (2-GOL). The prenyltransferase variants can be used to form prenylated aromatic compounds, and can be expressed in an engineered microbe having a pathway to such compounds, which include 3-GOLA, 3-GDVA, 3-GOSA, and CBG. 3-GOLA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions.

公开(公告)号：US20210254030A1

公开(公告)日：2021-08-19

申请号：US16978629

申请日：2019-03-08

Applicant: Genomatica, Inc.

Inventor： Michael A. Noble ,  Kevin G. Hoff ,  Anna Lechner ,  Harish Nagarajan

IPC: C12N9/10 ,  C12P7/42 ,  C12P7/22

Abstract: Described herein are prenyltransferases including non-natural variants thereof having at least one amino acid substitution as compared to its corresponding natural or unmodified prenyltransferases and that are capable of at least two-fold greater rate of formation of cannabinoids such as cannabigerolic acid, cannabigerovarinic acid, cannabigerorcinic acid, and cannabigerol, as compared to a wild type control. Prenyltransferase variants also demonstrated regioselectivity to desired cannabinoid isomers such as CDBA (3-GOLA), 3-GDVA, 3-GOSA, and CBG (2-GOL). The prenyltransferase variants can be used to form prenylated aromatic compounds, and can be expressed in an engineered microbe having a pathway to such compounds, which include 3-GOLA, 3-GDVA, 3-GOSA, and CBG. 3-GOLA can be used for the preparation of cannabigerol (CBG), which can be used in therapeutic compositions.

公开(公告)号：US20160237410A1

公开(公告)日：2016-08-18

申请号：US15027169

申请日：2014-10-03

Applicant: GENOMATICA, INC.

Inventor： Stefan Andrae ,  Michael Patrick Kuchinskas ,  Jingyi Li ,  Harish Nagarajan ,  Priti Pharkya

IPC: C12N9/04 ,  C12P7/24

Abstract: Described herein are non-natural NAD+-dependent alcohol dehydrogenases (ADHs) capable of at least two fold greater conversion of methanol or ethanol to formaldehyde or acetaldehyde, respectively, as compared to its unmodified counterpart. Nucleic acids encoding the non-natural alcohol dehydrogenases, as well as expression constructs including the nucleic acids, and engineered cells comprising the nucleic acids or expression constructs are described. Also described are engineered cells expressing a non-natural NAD+-dependent alcohol dehydrogenase, optionally include one or more additional metabolic pathway transgene(s), methanol metabolic pathway genes, target product pathway genes, cell culture compositions including the cells, methods for promoting production of the target product or intermediate thereof from the cells, compositions including the target product or intermediate, and products made from the target product or intermediate.

Abstract translation: 本文描述了与其未修饰的对应物相比，能够分别甲醇或乙醇转化为甲醛或乙醛的至少两倍的非天然NAD +依赖性醇脱氢酶（ADH）。 描述了编码非天然醇脱氢酶的核酸，以及包含核酸的表达构建体和包含核酸或表达构建体的工程化细胞。 还描述了表达非天然NAD +依赖性醇脱氢酶的工程细胞，任选地包括一种或多种另外的代谢途径转基因，甲醇代谢途径基因，靶产物途径基因，包括细胞的细胞培养组合物，用于促进生产的方法 的靶产物或其中间体，包括靶产物或中间体的组合物，以及由目标产物或中间体制成的产品。

公开(公告)号：US12234463B2

公开(公告)日：2025-02-25

申请号：US17255352

申请日：2019-06-26

Applicant: Genomatica, Inc.

Inventor： Harish Nagarajan ,  Tae Hoon Yang ,  Ali Khodayari

IPC: C12N9/16 ,  C12N9/02 ,  C12N9/88 ,  C12N9/90 ,  C12N15/52 ,  C12P5/00 ,  C12P5/02 ,  C12P7/16 ,  C12P7/18 ,  C12P7/24 ,  C12P7/42 ,  C12P7/46 ,  C12P7/6409 ,  C12P13/00

Abstract: Described herein are engineered cells including ones having synthetic methylotrophy which include an NADH-dependent enzyme capable of converting G3P to 3PG (e.g., B. methanolicus gapN) and/or fructose-1,6-bisphosphatase, along with hexulose-6-phosphate synthase, 6-phospho-3-hexuloisomerase, a phosphoketolase, or a combination thereof. Engineered cells of the disclosure beneficially maintain adequate pool sizes of phosphorylated C3 and/or C4 compounds, and/or provide increased levels of NADPH. As such, the modifications allow for the generation of C6 compounds from C1 (e.g., a methanol feedstod) and C5 compounds, the regeneration of C5 compounds from C6 compounds by carbon rearrangement, and an improved balance between regeneration of C5 compounds and lower glycolysis. In turn, this allows the engineered microorganism to generate sufficient quantities of metabolic precursors (e.g., acetyl-CoA) which can be used in a bioproduct pathway, and the engineered cells can include further modifications to those pathway enzymes allowing for production of a desired bioproduct.

公开(公告)号：US20220403420A1

公开(公告)日：2022-12-22

申请号：US17770233

申请日：2020-10-22

Applicant: Genomatica, Inc.

Inventor： Harish Nagarajan ,  Gionata Scalcinati ,  Tae Hoon Yang

IPC: C12P7/18 ,  C12N9/02 ,  C12N9/12 ,  C12N9/04

Abstract: The present disclosure provides microbial organisms having increased availability of co-factors, such as NADPH, for increasing production of various products, including 1,3-BDO, MMA, (3R)-hydroxybutyl (3R)-hydroxybutyrate, amino acids, 3HB-CoA, adipate, caprolactam, 6-ACA, HMD A, or MAA, and products made from any of these. Also provided are one or more exogenous nucleic acids encoding an enzyme expressed in a sufficient amount to increase availability of NADPH, where the exogenous nucleic acid includes one or more of ATP-NADH kinase, pntAB, nadK, and gapN. Also provided are one or more gene attenuations occurring in genes, such as NDH-2, that result in an increased ratio of NADPH to NADH. Various combinations of the exogenous nucleic acids and gene deletions are also provided in the present disclosure. The present disclosure also provides methods of making and using the same, including methods for culturing cells, and for the production of the various products.

公开(公告)号：US20220235385A1

公开(公告)日：2022-07-28

申请号：US17605196

申请日：2020-04-24

Applicant: Genomatica, Inc.

Inventor： Amit M. Shah ,  Harish Nagarajan

IPC: C12P13/00 ,  C12P17/08 ,  C12P17/10 ,  C12P7/18 ,  C12P7/44 ,  C12P7/62 ,  C12N9/02

Abstract: Disclosed are biosynthetic methods and engineered microorganism that enhance or improve the biosynthesis of hexamethylenediamine, caproic acid or caprolactam. The engineered microorganisms include selected aldehyde dehydrogenase activity.

公开(公告)号：US11384340B2

公开(公告)日：2022-07-12

申请号：US16792592

申请日：2020-02-17

Applicant: Genomatica, Inc.

Inventor： Stefan Andrae ,  Michael Patrick Kuchinskas ,  Jingyi Li ,  Harish Nagarajan ,  Priti Pharkya

IPC: C12N9/04 ,  C12P7/24 ,  C12P7/18

Abstract: Described herein are non-natural NAD+-dependent alcohol dehydrogenases (ADHs) capable of at least two fold greater conversion of methanol or ethanol to formaldehyde or acetaldehyde, respectively, as compared to its unmodified counterpart. Nucleic acids encoding the non-natural alcohol dehydrogenases, as well as expression constructs including the nucleic acids, and engineered cells comprising the nucleic acids or expression constructs are described. Also described are engineered cells expressing a non-natural NAD+-dependent alcohol dehydrogenase, optionally include one or more additional metabolic pathway transgene(s), methanol metabolic pathway genes, target product pathway genes, cell culture compositions including the cells, methods for promoting production of the target product or intermediate thereof from the cells, compositions including the target product or intermediate, and products made from the target product or intermediate.

公开(公告)号：US20200087634A1

公开(公告)日：2020-03-19

申请号：US16498336

申请日：2018-03-29

Applicant: Genomatica, Inc.

Inventor： Kevin Hoff ,  Cara Ann Tracewell ,  Kui Chan ,  Michael Kuchinkas ,  Harish Nagarajan

IPC: C12N9/04 ,  C12N9/10 ,  C12P19/32 ,  C12P7/52 ,  C12P7/24 ,  C12P7/18

Abstract: The present disclosure provides thiolases and polypeptide variants of 3-hydroxybutyryl-CoA dehydrogenase, nucleic acids encoding the same, vectors comprising the nucleic acids, and cells comprising the polypeptide variants and/or thiolase, the nucleic acids, and/or the vectors. The present disclosure also provides methods of making and using the same, including methods for culturing cells, and for the production of various products, including 3-hydroxybutyryl-CoA (3-HB-CoA), 3-hydroxybutyraldehyde (3-HBal), 3-hydroxybutyrate (3-HB), 1,3-butanediol (1,3-BDO), and esters and amides thereof, and products made from any of these.