公开(公告)号：WO2017075208A1

公开(公告)日：2017-05-04

申请号：PCT/US2016/059096

申请日：2016-10-27

Applicant: GENOMATICA, INC.

Inventor： BARTON, Nelson R. ,  LI, Jingyi ,  WARNER, Joseph R. ,  PHARKYA, Priti

IPC: C07K14/195 ,  C12N15/62 ,  C12N5/00

CPC classification number: C12Y503/01027 ,  C07K2319/00 ,  C12N9/0006 ,  C12N9/88 ,  C12N9/90 ,  C12N15/52 ,  C12Y401/02043

Abstract: Described herein are fusion proteins including methanol dehydrogenase (MeDH) and at least one other polypeptide such as 3-hexulose-6-phosphate dehydrogenase (HPS) or 6-phospho-3-hexuloisomerase (PHI), such as DHAS synthase or fructose-6-Phosphate aldolase or such as DHA synthase or DHA kinase. In a localized manner, the fusion protein can promote the conversion of methanol to formaldehyde and then to a ketose phosphate such as hexulose 6-phosphate or then to DHA and G3P. When expressed in cells, the fusion proteins can promote methanol uptake and rapid conversion to the ketose phosphate or to the DHA and D3P, which in turn can be used in a pathway for the production of a desired bioproduct. Beneficially, the rapid conversion to the ketose phosphate or to the DHA and G3P can avoid the undesirable accumulation of formaldehyde in the cell. Also described are engineered cells expressing the fusion protein, 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: 本文描述了融合蛋白，其包括甲醇脱氢酶（MeDH）和至少一种其他多肽如3-己酮糖-6-磷酸脱氢酶（HPS）或6-磷酸-3-己酮糖异构酶（PHI）， 如DHAS合酶或果糖-6-磷酸醛缩酶或例如DHA合酶或DHA激酶。 以局部方式，融合蛋白可以促进甲醇转化为甲醛，然后转化为磷酸酮糖如己酮糖6-磷酸，然后转化为DHA和G3P。 当在细胞中表达时，融合蛋白可以促进甲醇吸收并快速转化为磷酸酮糖或DHA和D3P，其又可用于生产期望的生物产品的途径中。 有利的是，快速转化为磷酸酮糖或DHA和G3P可以避免细胞中不希望的甲醛累积。 还描述了表达融合蛋白的工程细胞，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，用于促进目标产物或中间产物生成的方法 包括目标产物或中间体的组合物，以及由目标产物或中间体制备的产物。

公开(公告)号：EP3368548A1

公开(公告)日：2018-09-05

申请号：EP16791508.1

申请日：2016-10-27

Applicant: Genomatica, Inc.

Inventor： BARTON, Nelson R. ,  LI, Jingyi ,  WARNER, Joseph R. ,  PHARKYA, Priti

IPC: C07K14/195 ,  C12N15/62 ,  C12N5/00

CPC classification number: C12Y503/01027 ,  C07K2319/00 ,  C12N9/0006 ,  C12N9/88 ,  C12N9/90 ,  C12N15/52 ,  C12Y401/02043

Abstract: Described herein are fusion proteins including methanol dehydrogenase (MeDH) and at least one other polypeptide such as 3-hexulose-6-phosphate dehydrogenase (HPS) or 6-phospho-3-hexuloisomerase (PHI), such as DHAS synthase or fructose-6-Phosphate aldolase or such as DHA synthase or DHA kinase. In a localized manner, the fusion protein can promote the conversion of methanol to formaldehyde and then to a ketose phosphate such as hexulose 6-phosphate or then to DHA and G3P. When expressed in cells, the fusion proteins can promote methanol uptake and rapid conversion to the ketose phosphate or to the DHA and D3P, which in turn can be used in a pathway for the production of a desired bioproduct. Beneficially, the rapid conversion to the ketose phosphate or to the DHA and G3P can avoid the undesirable accumulation of formaldehyde in the cell. Also described are engineered cells expressing the fusion protein, 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.