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    • 5. 发明申请
    • BIOTECHNOLOGICAL PREPARATION OF 3-HYDROXYISOBUTYRIC ACID
    • 3-羟基丁酸的生物技术制备
    • US20150218600A1
    • 2015-08-06
    • US14363178
    • 2012-11-14
    • Thomas HaasSteffen SchafferMarkus PoetterMirja WesselJan Christoph PfefferChristian GehringNicole KirchnerEva Maria Wittmann
    • Thomas HaasSteffen SchafferMarkus PoetterMirja WesselJan Christoph PfefferChristian GehringNicole KirchnerEva Maria Wittmann
    • C12P7/42C12N9/88C12N9/12C12N9/02C12N9/00C12N9/10
    • C12P7/42C12N9/0004C12N9/001C12N9/1217C12N9/13C12N9/88C12N9/93C12N15/52C12Y103/99002C12Y207/02007C12Y402/01017C12Y602/01002
    • The invention relates to a method comprising the steps a) providing isobutyric acid, b) bringing isobutyric acid into contact with the combination of isobutyrate kinase and phosphotransisobutyrylase and/or isobutyryl-coenzyme A synthetase/ligase and/or isobutyrate-coenzyme A transferase, c) bringing the product from step a) into contact with isobutyryl-coenzyme A dehydrogenase, d) bringing the product from step b) into contact with methacrylyl-coenzyme A hydratase, and e) hydrolyzing the product from step d) to form 3-hydroxyisobutyric acid, where at least one of the enzymes is used in the form of a cell which, compared to its wildtype, comprises a reduced activity of a 3-hydroxyisobutyric acid dehydrogenase or a variant thereof, a cell which has at least one enzyme from the group comprising isobutyryl-coenzyme A synthetase/ligase, isobutyrate-coenzyme A transferase, isobutyrate kinase, phosphotransisobutyrylase, isobutyryl-coenzyme A dehydrogenase, methacrylyl-coenzyme A hydratase and 3-hydroxyisobutyryl-coenzyme A hydrolase and, compared to its wildtype, a reduced activity of a 3-hydroxyisobutyric acid dehydrogenase or a variant thereof, wherein the cell preferably has, in addition, a monooxygenase, more preferably a monooxygenase of the alkBGT type or a variant thereof and the use of such a cell for preparing 3-hydroxyisobutyric acid.
    • 本发明涉及包括以下步骤的方法:a)提供异丁酸,b)使异丁酸与异丁酸酯激酶和磷酸转移异丁酰基酶和/或异丁酰辅酶A合成酶/连接酶和/或异丁酸酯辅酶A转移酶c的组合接触 )使来自步骤a)的产物与异丁酰辅酶A脱氢酶接触,d)将来自步骤b)的产物与甲基丙烯酰辅酶A水合酶接触,和e)水解步骤d)中的产物以形成3-羟基异丁酸 酸,其中至少一种酶以细胞形式使用,与其野生型相比,其包含3-羟基异丁酸脱氢酶或其变体的降低的活性,具有至少一种酶的细胞 异丁酰辅酶A合成酶/连接酶,异丁酸酯辅酶A转移酶,异丁酸酯激酶,磷酸转移异丁酰基酶,异丁酰辅酶A脱氢酶,甲基丙烯酰辅酶A水合酶和 3-羟基异丁酰辅酶A水解酶,与其野生型相比,3-羟基异丁酸脱氢酶或其变体的活性降低,其中细胞优选还具有单加氧酶,更优选alkBGT型的单加氧酶或 其变体以及这种细胞用于制备3-羟基异丁酸的用途。