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    Method for the in vivo modification of the synthesis activity of a metabolite by means of the modification of a gene the activity of which is not the original activity
    3.
    发明授权
    Method for the in vivo modification of the synthesis activity of a metabolite by means of the modification of a gene the activity of which is not the original activity 有权
    通过改变基因的体内修饰代谢物的合成活性的方法，其活性不是原始活性

    公开(公告)号：US07892838B2

    公开(公告)日：2011-02-22

    申请号：US11735148

    申请日：2007-04-13

    申请人： Pierre-Alexandre Kaminski ,  Philippe Marliere

    发明人： Pierre-Alexandre Kaminski ,  Philippe Marliere

    IPC分类号： C12P21/06 ,  C12P19/30 ,  C12N9/10 ,  C12N1/20 ,  C12N15/00 ,  C12N15/74 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C12N15/1058 ,  C12N15/102

    摘要： The invention relates to a method for altering a protein X such as to modify the characteristics thereof by a) obtaining the mutants X* of the sequence coding for protein X, by means of aleatory mutagenesis, b) transformation of cells with a phenotype [P-] with vectors comprising the mutated nucleic acids obtained in step (a) which code for proteins X*, where P-signifies that said cells are auxotrophic for substance P, P begin the product of the action of X on the natural substrate thereof S, c) culturing said cells in a medium comprising a substrate S*, S* being an analogue of the natural substrate S of the protein X, d) selection of the cells [P-:: X*] which have survived step c) in which the proteins X* can biosynthesise the product P from the substrate S*. The invention further relates to mutated proteins X, nucleic acids, expression vectors, host cells comprising a vector, use of N-dideoxyribosyl transferases for the transfer of a dideoxyribose (ddR) from a dideoxyribonucleoside to another nucleoside, a method for production of compounds comprising a step using a mutated protein and a strain of E. coli.

    摘要翻译： 本发明涉及一种用于改变蛋白质X的方法，例如通过以下方式改变其特征：a）通过亲和诱变获得编码蛋白质X的序列的突变体X *，b）用表型转化的细胞[P - ]，其中包含步骤（a）中获得的编码蛋白质X *的突变核酸的载体，其中P表示所述细胞对于物质P是营养缺陷型的，P开始在其天然底物上的作用X的产物S c）在包含蛋白质X的天然底物S的类似物的底物S *，S *的培养基中培养所述细胞，d）选择在步骤c）中存活的细胞[P- :: X *]， 其中蛋白质X *可以从底物S *生物合成产物P. 本发明进一步涉及突变蛋白X，核酸，表达载体，包含载体的宿主细胞，使用N-二脱氧核糖基转移酶将二脱氧核糖（ddR）从双脱氧核糖核苷转移至另一核苷的方法，包括 使用突变蛋白和大肠杆菌菌株的步骤。

    METHOD FOR THE IN VIVO MODIFICATION OF THE SYNTHESIS ACTIVITY OF A METABOLITE BY MEANS OF THE MODIFICATION OF A GENE THE ACTIVITY OF WHICH IS NOT THE ORIGINAL ACTIVITY
    5.
    发明申请
    METHOD FOR THE IN VIVO MODIFICATION OF THE SYNTHESIS ACTIVITY OF A METABOLITE BY MEANS OF THE MODIFICATION OF A GENE THE ACTIVITY OF WHICH IS NOT THE ORIGINAL ACTIVITY 有权
    通过修饰基因的方法修饰代谢物的合成活性的方法不是原始活性的活性

    公开(公告)号：US20080299608A1

    公开(公告)日：2008-12-04

    申请号：US11735148

    申请日：2007-04-13

    申请人： Pierre-Alexandre Kaminski ,  Philippe Marliere

    发明人： Pierre-Alexandre Kaminski ,  Philippe Marliere

    IPC分类号： C12P21/04 ,  C12N15/87 ,  C12N9/10 ,  C12N15/00 ,  C12P19/30 ,  C07H21/00 ,  C12N1/20 ,  C12N15/11 ,  C12N15/63

    CPC分类号： C12N15/1058 ,  C12N15/102

    摘要： The invention relates to a method for altering a protein X such as to modify the characteristics thereof by a) obtaining the mutants X* of the sequence coding for protein X, by means of aleatory mutagenesis, b) transformation of cells with a phenotype [P-] with vectors comprising the mutated nucleic acids obtained in step (a) which code for proteins X*, where P-signifies that said cells are auxotrophic for substance P, P begin the product of the action of X on the natural substrate thereof S, c) culturing said cells in a medium comprising a substrate S*, S* being an analogue of the natural substrate S of the protein X, d) selection of the cells [P-:: X*] which have survived step c) in which the proteins X* can biosynthesise the product P from the substrate S*. The invention further relates to mutated proteins X, nucleic acids, expression vectors, host cells comprising a vector, use of N-dideoxyribosyl transferases for the transfer of a dideoxyribose (ddR) from a dideoxyribonucleoside to another nucleoside, a method for production of compounds comprising a step using a mutated protein and a strain of E. coli.

    摘要翻译： 本发明涉及一种用于改变蛋白质X的方法，例如通过以下方式改变其特征：a）通过亲和诱变获得编码蛋白质X的序列的突变体X *，b）用表型转化的细胞[P - ]，其中包含步骤（a）中获得的编码蛋白质X *的突变核酸的载体，其中P表示所述细胞对于物质P是营养缺陷型的，P开始在其天然底物上的作用X的产物S c）在包含蛋白质X的天然底物S的类似物的底物S *，S *的培养基中培养所述细胞，d）选择在步骤c）中存活的细胞[P- :: X *]， 其中蛋白质X *可以从底物S *生物合成产物P. 本发明进一步涉及突变蛋白X，核酸，表达载体，包含载体的宿主细胞，使用N-二脱氧核糖基转移酶将二脱氧核糖（ddR）从双脱氧核糖核苷转移至另一核苷的方法，包括 使用突变蛋白和大肠杆菌菌株的步骤。

    Lactobacillus fermentum N-desoxyribosyl transferases and the use thereof for enzymatic synthesis of 2′, 3′—didesoxynucleosides and 2′,3′- didehydro-2′,3′- didesoxynucleosides
    8.
    发明授权
    Lactobacillus fermentum N-desoxyribosyl transferases and the use thereof for enzymatic synthesis of 2′, 3′—didesoxynucleosides and 2′,3′- didehydro-2′,3′- didesoxynucleosides 有权
    发酵乳杆菌N-脱氧核糖基转移酶及其用于酶促合成2'，3'-二氧代核苷和2'，3'-二脱氢-2'，3'-二硫代核苷的用途

    公开(公告)号：US07820795B2

    公开(公告)日：2010-10-26

    申请号：US10594766

    申请日：2005-03-29

    申请人： Pierre-Alexandre Kaminski

    发明人： Pierre-Alexandre Kaminski

    IPC分类号： A23J1/18 ,  C12Q1/48 ,  C12N15/63 ,  C12N15/85 ,  C12N15/11

    CPC分类号： C12N9/1077

    摘要： N-deoxyribosyl transferases of Lactobacillus fermentum and their analogues, as well their use for the enzymatic synthesis of 2′,3′-dideoxynucleosides and 2′,3′-didehydro-2′,3′-dideoxynucleosides. These transferases and their analogues include a N-deoxyribosyl transferase protein (DTP) that has at least 70%-95% identity with the polypeptide of SEQ ID NO: 2 or SEQ ID NO: 4, that retains residues Y13, D77, D97, E103, and M312 which respectively correspond to positions 13, 77, 97, 103, and 132 of SEQ ID NO: 2; and that has threonine at a position corresponding to position 15 of SEQ ID NO: 2 or SEQ ID NO: 4. Polynucleotides, vectors and host cells encoding these N-deoxyribosyl transferases and their analogues.

    摘要翻译： 发酵乳杆菌的N-脱氧核糖基转移酶及其类似物，以及它们用于酶促合成2'，3'-二脱氧核苷和2'，3'-二脱氢-2'，3'-双脱氧核苷的用途。 这些转移酶及其类似物包括与SEQ ID NO：2或SEQ ID NO：4的多肽具有至少70％-95％同一性的N-脱氧核糖基转移酶蛋白（DTP），其保留残基Y13，D77，D97， E103和M312分别对应于SEQ ID NO：2的位置13,77,97,103和132; 并且在对应于SEQ ID NO：2或SEQ ID NO：4的位置15的位置具有苏氨酸。编码这些N-脱氧核糖基转移酶及其类似物的多核苷酸，载体和宿主细胞。

    Lactobacillus Fermentum N-Desoxyribosyl Transferases and the Use Thereof for Enzymatic Synthesis of 2', 3' - Didesoxynucleosides and 2',3'- Didehydro-2',3'- Didesoxynucleosides
    9.
    发明申请
    Lactobacillus Fermentum N-Desoxyribosyl Transferases and the Use Thereof for Enzymatic Synthesis of 2', 3' - Didesoxynucleosides and 2',3'- Didehydro-2',3'- Didesoxynucleosides 有权
    发酵酵母N-脱氧核糖转移酶及其用途用于2'，3'-二硫代核苷和2'，3'-二脱氢-2'，3'-二硫代核苷的酶合成

    公开(公告)号：US20080064646A1

    公开(公告)日：2008-03-13

    申请号：US10594766

    申请日：2005-03-29

    申请人： Pierre-Alexandre Kaminski

    发明人： Pierre-Alexandre Kaminski

    IPC分类号： A61K31/70 ,  C07H21/04 ,  C12N15/00 ,  C12N5/04 ,  C12P19/38

    CPC分类号： C12N9/1077

    摘要： The inventive method for evaluating an X protein encoded by an Lactobacillus fermentum (L. fermentum ) ntd gene in such a way that the characteristics thereof are modified consists a) in obtaining the Lactobacillus fermentum (L. fermentum ) ntd gene mutants by random mutagenesis, b) in transforming cells containing a [P-] phenotype provided with vectors containing mutated nucleic acids obtained at the stage a) coding for the thus modified X* proteins, wherein P- means that said cells are auxotrophic for a substance P produced by the action of X on a natural substrate S, c) in culturing said cells in a medium comprising a substrate S*, wherein S* is an analog to the natural substrate S of the protein X and d) in selecting the cells [P-::X*] which survived at the stage c) and ijn which the proteins X* are capable of carrying out the biosynthesis of the product P based on the substrate S*. The mutated L. fermentum N-desoxyribosyl transferases have an N-didesoxyribosyl transferase activity, corresponding nucleic acids, expression vectors, host cells containing said vectors and an application for the enzymatic synthesis of 2′,3′-didesoxynucleosides and 2′,3′-didehydro-2′,3′-didesoxynucleosides.

    摘要翻译： 本发明的由发酵乳杆菌（Lactobacillus fermentum）（L. fermentum）ntd基因编码的X蛋白的方法，其特征被修饰的方法是a）通过随机诱变获得发酵乳杆菌（Lactobacillus fermentum）（L. fermentum）ntd基因突变体， b）在转化含有[P-]表型的细胞中，所述细胞含有在步骤a）获得的含有突变的核酸的载体，编码如此修饰的X *蛋白质，其中P-表示所述细胞对于所产生的物质P产生营养缺陷型 X在天然底物S上的作用，c）在包含底物S *的培养基中培养所述细胞，其中S *与蛋白X的天然底物S类似，d）在选择细胞[P-： ：X *]，其在蛋白质X *能够基于底物S *进行产物P的生物合成的阶段c）和ijn中存活。 突变的发酵酵母N-脱氧核糖基转移酶具有N-二硫代氧代核糖基转移酶活性，相应的核酸，表达载体，含有所述载体的宿主细胞和用于酶促合成2'，3'-二硫代核苷和2'，3' - 二脱氢-2'，3'-二硫代核苷。