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1. 发明授权

US07544498B2 Aromatic prenyltransferases, nucleic acids encoding same and uses therefor 有权
标题翻译：芳族异戊烯基转移酶，编码相同的核酸，用于其
公开(公告)号：US07544498B2
公开(公告)日：2009-06-09
申请号：US12106181
申请日：2008-04-18
申请人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
发明人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
IPC分类号： G01N33/573 , C12N9/10 , C12P1/00
CPC分类号： C12N9/1085
摘要： In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved in naphterpin biosynthesis has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg2+, and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture. The complexes with both aromatic substrates and prenyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and aromatic co-substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.
摘要翻译：根据本发明，一种新型芳族异戊烯基转移酶，来自链霉菌属的Orf2。已经鉴定了参与萘普生生物合成的菌株CL190，其结构被阐明。该异戊烯基转移酶催化异戊烯基和含有芳香核的化合物之间的C-C键的形成，并且还显示C-O键的形成活性。已经解决和改进了异戊烯基转移酶的许多晶体结构，例如（1）与缓冲分子（TAPS）复合的异戊烯基转移酶，（2）作为二进位配合物的前体甲基转移酶与香叶基二磷酸（GPP）和Mg2 +以及异戊烯基转移酶作为三元复合物不可水解的底物类似物，香叶基S-硫代双磷酸（GSPP）和（3）1,6-二羟基萘（1,6-DHN）或（4）黄堇素（即2,5,7-三羟基-1， 4-萘醌，其为1,3,6,8-四羟基萘（THN）的氧化产物）。这些结构分别被解析和精炼成1.5A，2.25，1.95和2.02。芳族异戊烯基转移酶的这种第一结构显示出意想不到的和非规范（β/α） - 杆状结构。具有芳香底物和含异戊烯基的底物和类似物的络合物描绘了活性位点，并与提出的异戊烯基转移的亲电子机制一致。这些结构还提供了在这个结构独特的芳族异戊烯基转移酶家族中了解异戊烯基链长度测定和芳香族共底物识别的机理基础。该结构信息对于预测蛋白质的芳族异戊烯基转移酶活性是有用的。

2. 发明申请

US20100285502A1 NOVEL AROMATIC PRENYLTRANSFERASES, NUCLEIC ACIDS ENCODING SAME AND USES THEREFOR 有权
标题翻译：新型芳香丙烯酸酯，编码其的核酸及其用途
公开(公告)号：US20100285502A1
公开(公告)日：2010-11-11
申请号：US12480618
申请日：2009-06-08
申请人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
发明人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
IPC分类号： G01N33/573 , C12P21/06 , C12N9/10 , C07H21/04
CPC分类号： C12N9/1085
摘要： In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved in naphterpin biosynthesis has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg2+, and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture. The complexes with both aromatic substrates and prenyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and aromatic co-substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.
摘要翻译：根据本发明，一种新型芳族异戊烯基转移酶，来自链霉菌属的Orf2。已经鉴定了参与萘普生生物合成的菌株CL190，其结构被阐明。该异戊烯基转移酶催化异戊烯基和含有芳香核的化合物之间的C-C键的形成，并且还显示C-O键的形成活性。已经解决和改进了异戊烯基转移酶的许多晶体结构，例如（1）与缓冲分子（TAPS）复合的异戊烯基转移酶，（2）作为二进位配合物的前体甲基转移酶与香叶基二磷酸（GPP）和Mg2 +以及异戊烯基转移酶作为三元复合物不可水解的底物类似物，香叶基S-硫代双磷酸（GSPP）和（3）1,6-二羟基萘（1,6-DHN）或（4）黄堇素（即2,5,7-三羟基-1， 4-萘醌，其为1,3,6,8-四羟基萘（THN）的氧化产物）。这些结构分别被解析和精炼成1.5A，2.25，1.95和2.02。芳香族异戊烯基转移酶的这种第一种结构显示出意想不到的和非规范的（＆bgr。/α） - 桶结构。具有芳香底物和含异戊烯基的底物和类似物的络合物描绘了活性位点，并与提出的异戊烯基转移的亲电子机制一致。这些结构还提供了在这个结构独特的芳族异戊烯基转移酶家族中了解异戊烯基链长度测定和芳香族共底物识别的机理基础。该结构信息对于预测蛋白质的芳族异戊烯基转移酶活性是有用的。

3. 发明授权

US08124390B2 Aromatic prenyltransferases, nucleic acids encoding same and uses therefor 有权
标题翻译：芳族异戊烯基转移酶，编码相同的核酸，用于其
公开(公告)号：US08124390B2
公开(公告)日：2012-02-28
申请号：US12480618
申请日：2009-06-08
申请人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
发明人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
IPC分类号： G01N33/573 , C12N9/10 , C12P1/00
CPC分类号： C12N9/1085
摘要： In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved in naphterpin biosynthesis has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg2+, and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture. The complexes with both aromatic substrates and prenyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and aromatic co-substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.
摘要翻译：根据本发明，一种新型芳族异戊烯基转移酶，来自链霉菌属的Orf2。已经鉴定了参与萘普生生物合成的菌株CL190，其结构被阐明。该异戊烯基转移酶催化异戊烯基和含有芳香核的化合物之间的C-C键的形成，并且还显示C-O键的形成活性。已经解决和改进了异戊烯基转移酶的许多晶体结构，例如（1）与缓冲分子（TAPS）复合的异戊烯基转移酶，（2）作为二进位配合物的前体甲基转移酶与香叶基二磷酸（GPP）和Mg2 +以及异戊烯基转移酶作为三元复合物不可水解的底物类似物，香叶基S-硫代双磷酸（GSPP）和（3）1,6-二羟基萘（1,6-DHN）或（4）黄堇素（即2,5,7-三羟基-1， 4-萘醌，其为1,3,6,8-四羟基萘（THN）的氧化产物）。这些结构分别被解析和精炼成1.5A，2.25，1.95和2.02。芳香族异戊烯基转移酶的这种第一种结构显示出意想不到的和非规范的（＆bgr。/α） - 桶结构。具有芳香底物和含异戊烯基的底物和类似物的络合物描绘了活性位点，并与提出的异戊烯基转移的亲电子机制一致。这些结构还提供了在这个结构独特的芳族异戊烯基转移酶家族中了解异戊烯基链长度测定和芳香族共底物识别的机理基础。该结构信息对于预测蛋白质的芳族异戊烯基转移酶活性是有用的。

4. 发明申请

US20080274478A1 NOVEL AROMATIC PRENYLTRANSFERASES, NUCLEIC ACIDS ENCODING SAME AND USES THEREFOR 有权
标题翻译：新型芳香丙烯酸酯，编码其的核酸及其用途
公开(公告)号：US20080274478A1
公开(公告)日：2008-11-06
申请号：US12106181
申请日：2008-04-18
申请人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
发明人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
IPC分类号： G01N33/573 , C12N9/10 , C12P1/00
CPC分类号： C12N9/1085
摘要： In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved in naphterpin biosynthesis has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg2+, and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture. The complexes with both aromatic substrates and prenyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and aromatic co-substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.
摘要翻译：根据本发明，一种新型芳族异戊烯基转移酶，来自链霉菌属的Orf2。已经鉴定了参与萘普生生物合成的菌株CL190，其结构被阐明。该异戊烯基转移酶催化异戊烯基和含有芳香核的化合物之间的C-C键的形成，并且还显示C-O键的形成活性。已经解决和改进了异戊烯基转移酶的许多晶体结构，例如（1）与缓冲分子（TAPS）复合的异戊烯基转移酶，（2）作为二进位配合物的异戊烯基转移酶与香叶基二磷酸酯（GPP）和Mg 2+ （3）1,6-二羟基萘（1,6-DHN）或（4）黄堇素（即2号）的三钠配合物，，5,7-三羟基-1,4-萘醌，其为1,3,6,8-四羟基萘（THN）的氧化产物）。这些结构分别被解析和精炼成1.5A，2.25，1.95和2.02。芳族异戊烯基转移酶的这种第一结构显示出意想不到的和非规范（β/α） - 杆状结构。具有芳香底物和含异戊烯基的底物和类似物的络合物描绘了活性位点，并与提出的异戊烯基转移的亲电子机制一致。这些结构还提供了在这个结构独特的芳族异戊烯基转移酶家族中了解异戊烯基链长度测定和芳香族共底物识别的机理基础。该结构信息对于预测蛋白质的芳族异戊烯基转移酶活性是有用的。

5. 发明授权

US07361483B2 Aromatic prenyltransferases, nucleic acids encoding same and uses therefor 有权
标题翻译：芳族异戊烯基转移酶，编码相同的核酸，用于其
公开(公告)号：US07361483B2
公开(公告)日：2008-04-22
申请号：US11342328
申请日：2006-01-27
申请人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
发明人： Tomohisa Kuzuyama , Joseph P. Noel , Stephane P. Richard
IPC分类号： C12N9/10 , C12Q1/48
CPC分类号： C12N9/1085
摘要： In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved in naphterpin biosynthesis has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg2+, and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture. The complexes with both aromatic substrates and prenyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and aromatic co-substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.
摘要翻译：根据本发明，一种新型芳族异戊烯基转移酶，来自链霉菌属的Orf2。已经鉴定了参与萘普生生物合成的菌株CL190，其结构被阐明。该异戊烯基转移酶催化异戊烯基和含有芳香核的化合物之间的C-C键的形成，并且还显示C-O键的形成活性。已经解决和改进了异戊烯基转移酶的许多晶体结构，例如（1）与缓冲分子（TAPS）复合的异戊烯基转移酶，（2）作为二进位配合物的异戊烯基转移酶与香叶基二磷酸酯（GPP）和Mg 2+ （3）1,6-二羟基萘（1,6-DHN）或（4）黄堇素（即2号）的三钠配合物，，5,7-三羟基-1,4-萘醌，其为1,3,6,8-四羟基萘（THN）的氧化产物）。这些结构分别被解析和精炼成1.5A，2.25，1.95和2.02。芳族异戊烯基转移酶的这种第一结构显示出意想不到的和非规范（β/α） - 杆状结构。具有芳香底物和含异戊烯基的底物和类似物的络合物描绘了活性位点，并与提出的异戊烯基转移的亲电子机制一致。这些结构还提供了在这个结构独特的芳族异戊烯基转移酶家族中了解异戊烯基链长度测定和芳香族共底物识别的机理基础。该结构信息对于预测蛋白质的芳族异戊烯基转移酶活性是有用的。

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