公开(公告)号：US09006412B2

公开(公告)日：2015-04-14

申请号：US13122675

申请日：2009-10-05

申请人： Yoshikazu Fujii ,  Tadashi Fujii ,  Akira Arisawa ,  Tomohiro Tamura

发明人： Yoshikazu Fujii ,  Tadashi Fujii ,  Akira Arisawa ,  Tomohiro Tamura

IPC分类号： C07H21/04 ,  C12N15/00 ,  C12N15/64 ,  C12P21/06 ,  C12P21/02 ,  C12N9/02 ,  C12N15/74 ,  C12P7/02 ,  C12P7/62 ,  C12P17/06

CPC分类号： C12P21/02 ,  C12N9/0071 ,  C12N9/0073 ,  C12N15/74 ,  C12P7/02 ,  C12P7/62 ,  C12P17/06 ,  C12Y114/13013

摘要： An expression vector capable of expressing a foreign gene in Pseudonocardia autotrophica; a transformant of Pseudonocardia autotrophica produced by using the expression vector; a method for producing a protein by using the transformant; a method for producing an active form of vitamin D3 from vitamin D3, which comprises highly expressing a gene encoding an enzyme involved in the synthesis of the active form of vitamin D3 in a transformant by using the expression vector or the transformant; a method for producing 25-hydroxyvitamin D2 from vitamin D2; and a method for producing pravastatin from compactin, which comprises highly expressing a compactin hydroxylase gene in a transformant by using the expression vector or the transformant.

摘要翻译： 能够在自养假诺卡氏菌中表达外源基因的表达载体; 通过使用表达载体产生的假心内膜自养基因的转化体; 通过使用该转化体生产蛋白质的方法; 从维生素D3生产活性形式的维生素D3的方法，其包括通过使用表达载体或转化体高度表达编码参与转化体中活性形式的维生素D3的活性形式的酶的基因; 从维生素D2生产25-羟基维生素D 2的方法; 以及通过使用表达载体或转化体高效表达转化体中的致密蛋白羟化酶基因的来自美伐他汀的普伐他汀的制造方法。

公开(公告)号：US20110262978A1

公开(公告)日：2011-10-27

申请号：US13122675

申请日：2009-10-05

申请人： Yoshikazu Fujii ,  Tadashi Fujii ,  Akira Arisawa ,  Tomohiro Tamura

发明人： Yoshikazu Fujii ,  Tadashi Fujii ,  Akira Arisawa ,  Tomohiro Tamura

IPC分类号： C12P7/02 ,  C12N15/74 ,  C12N9/02 ,  C12P7/62 ,  C12N15/63 ,  C12N1/21

CPC分类号： C12P21/02 ,  C12N9/0071 ,  C12N9/0073 ,  C12N15/74 ,  C12P7/02 ,  C12P7/62 ,  C12P17/06 ,  C12Y114/13013

摘要： An expression vector capable of expressing a foreign gene in Pseudonocardia autotrophica; a transformant of Pseudonocardia autotrophica produced by using the expression vector; a method for producing a protein by using the transformant; a method for producing an active form of vitamin D3 from vitamin D3, which comprises highly expressing a gene encoding an enzyme involved in the synthesis of the active form of vitamin D3 in a transformant by using the expression vector or the transformant; a method for producing 25-hydroxyvitamin D2 from vitamin D2; and a method for producing pravastatin from compactin, which comprises highly expressing a compactin hydroxylase gene in a transformant by using the expression vector or the transformant.

摘要翻译： 能够在自养假诺卡氏菌中表达外源基因的表达载体; 通过使用表达载体产生的假心内膜自养基因的转化体; 通过使用该转化体生产蛋白质的方法; 从维生素D3生产活性形式的维生素D3的方法，其包括通过使用表达载体或转化体高度表达编码参与转化体中活性形式的维生素D3的活性形式的酶的基因; 从维生素D2生产25-羟基维生素D 2的方法; 以及通过使用表达载体或转化体高效表达转化体中的致密蛋白羟化酶基因的来自美伐他汀的普伐他汀的制造方法。

公开(公告)号：US20120178124A1

公开(公告)日：2012-07-12

申请号：US13143607

申请日：2009-01-07

申请人： Masahiro Yamagishi ,  Takeshi Sakamoto ,  Makoto Ueda ,  Masashi Itoh ,  Yoshikazu Fujii ,  Hiroki Kabumoto ,  Akira Arisawa ,  Tadashi Fujii

发明人： Masahiro Yamagishi ,  Takeshi Sakamoto ,  Makoto Ueda ,  Masashi Itoh ,  Yoshikazu Fujii ,  Hiroki Kabumoto ,  Akira Arisawa ,  Tadashi Fujii

IPC分类号： C12P33/00 ,  C12N15/52 ,  C12N1/15 ,  C12N9/96 ,  C12N1/21 ,  C12N1/19 ,  C12N9/00 ,  C12N15/63

CPC分类号： C12N9/0077 ,  C12N9/0081 ,  C12N15/70 ,  C12N15/75 ,  C12P33/12

摘要： It is an object of the present invention to obtain highly active P450scc enzyme protein which is an important enzyme protein that catalyzes the first step of the biosynthesis of industrially useful steroid hormone. The present invention provides a sterol side chain cleavage enzyme protein having the following physicochemical properties: (1) action: the enzyme acts on sterol represented by formula (I) as defined in the specification and cleaves the carbon-carbon bond between positions 20 and 22 of a sterol side chain portion by its activity of cleaving the bonds, so as to generate a compound represented by formula (II) as defined in the specification; (2) substrate specificity: when microorganisms that produce the enzyme protein are allowed to react with an aqueous solution containing 100 μg/ml 4-cholesten-3-one or cholesterol at 28° C. for 5 hours, the conversion reaction rate from 4-cholesten-3-one to progesterone is 10% or more, and the conversion rate from cholesterol to pregnenolone is 10% or more;

摘要翻译： 本发明的目的是获得高活性P450scc酶蛋白，其是催化工业上有用的类固醇激素生物合成的第一步的重要酶蛋白。 本发明提供了具有以下物理化学性质的甾醇侧链切割酶蛋白：（1）作用：酶作用于如说明书中定义的由式（I）表示的固醇，并切割位置20和22之间的碳 - 碳键 的甾醇侧链部分通过其切割键的活性，从而产生如说明书中定义的式（II）表示的化合物; （2）底物特异性：使得产生酶蛋白的微生物在28℃下与含有100μg/ ml 4-胆甾烯-3-酮或胆固醇的水溶液反应5小时时，转化反应速率为4 胆固醇-3-酮与孕酮的含量为10％以上，胆固醇与孕烯醇酮的转化率为10％以上;

公开(公告)号：US6143541A

公开(公告)日：2000-11-07

申请号：US16

申请日：1998-01-30

申请人： Akira Arisawa ,  Motoko Matsufuji ,  Takurou Tsuruta ,  Kazuyuki Dobashi ,  Takashi Nakashima ,  Kunio Isshiki ,  Takeo Yoshioka

发明人： Akira Arisawa ,  Motoko Matsufuji ,  Takurou Tsuruta ,  Kazuyuki Dobashi ,  Takashi Nakashima ,  Kunio Isshiki ,  Takeo Yoshioka

IPC分类号： C07D211/88 ,  C07D211/90 ,  C12N9/18 ,  C12N9/78 ,  C12N15/55 ,  C12P17/12 ,  C12P41/00 ,  C12N9/14 ,  C12N1/20 ,  C12N9/16 ,  C12P12/12

CPC分类号： C07D211/88 ,  C07D211/90 ,  C12N9/18 ,  C12N9/78 ,  C12P17/12 ,  C12P41/006

摘要： This invention provides DNA fragments for efficiently preparing a protein having asymmetric hydrolase activity for 4-substituted 1,4-dihydropyridine derivatives, transformants obtained by using such a DNA fragment, and a process for preparing the aforesaid enzyme. Specifically, an isolated gene derived from the chromosome of Streptomyces viridosporus and encoding a protein having asymmetric hydrolase activity for 4-substituted 1,4-dihydropyridine derivatives, plasmids having the aforesaid gene integrated thereinto, transformants obtained by using such a plasmid, and a process for preparing the aforesaid enzyme by using such a transformant are disclosed. A process for the enzymatic conversion of 4-substituted 1,4-dihydropyridine derivatives by using the aforesaid enzyme is also disclosed.

摘要翻译： PCT No.PCT / JP96 / 02147 Sec。 371日期1998年1月30日 102（e）1998年1月30日PCT PCT 1996年7月30日PCT公布。 公开号WO97 / 05243 日期1997年2月13日本发明提供了用于有效制备对4-取代的1,4-二氢吡啶衍生物具有不对称水解酶活性的蛋白质的DNA片段，以及通过使用该DNA片段得到的转化体及其制备方法。 具体来说，来源于链球菌链球菌的染色体的分离的基因，编码对4-取代的1,4-二氢吡啶衍生物具有不对称的水解酶活性的蛋白质，将上述基因整合到其中的质粒，使用这样的质粒获得的转化体， 公开了通过使用这种转化体制备上述酶的方法。 还公开了通过使用上述酶酶促转化4-取代的1,4-二氢吡啶衍生物的方法。

公开(公告)号：US20090269820A1

公开(公告)日：2009-10-29

申请号：US11630689

申请日：2005-07-19

申请人： Kazuhiro Machida ,  Akira Arisawa ,  Susumu Takeda ,  Masashi Yoshida ,  Toshio Tsuchida

发明人： Kazuhiro Machida ,  Akira Arisawa ,  Susumu Takeda ,  Masashi Yoshida ,  Toshio Tsuchida

IPC分类号： C12P17/16 ,  C07H21/04 ,  C07K2/00 ,  C12N9/00 ,  C12N15/63 ,  C12N1/00

CPC分类号： C12P17/08 ,  C07K14/36 ,  C12N9/0071 ,  C12N9/0073 ,  C12N15/52 ,  C12P17/162

摘要： The present invention provides polypeptides that participate in the biosynthesis of the pladienolide macrolide compounds, DNA that encodes these polypeptides and variants of this DNA, transformants that maintain all or a portion of this DNA or variant thereof, and a method of producing the pladienolide macrolide compounds using these transformants. More particularly, it provides an isolated pure DNA that contains at least one region encoding a polypeptide that participates in pladienolide biosynthesis; polypeptide encoded by this DNA; a self-replicating or integrated-replicating recombinant plasmid carrying this DNA; a transformant maintaining this DNA; and a method of producing a pladienolide, characterized by culturing this transformant on culture medium and collecting pladienolide from this culture medium.

摘要翻译： 本发明提供了参与扁平二烯内酰脲大环内酯化合物的生物合成的多肽，编码这些多肽的DNA和该DNA的变体，保持该DNA或其变体的全部或一部分的转化体，以及生产该扁桃苷内酰胺大环内酯化合物的方法 使用这些转化体。 更具体地，其提供了分离的纯DNA，其含有至少一个编码参与泛二烯内酯生物合成的多肽的区域; 由该DNA编码的多肽; 携带该DNA的自我复制或整合复制重组质粒; 维持该DNA的转化体 以及生产普拉二烯内酯的方法，其特征在于在培养基上培养该转化体并从该培养基中收集普拉二烯内酯。

公开(公告)号：US5240849A

公开(公告)日：1993-08-31

申请号：US356323

申请日：1989-05-23

申请人： Akira Arisawa ,  Naoto Kawamura ,  Ikuo Kojima ,  Yasushi Okumura ,  Kazuhiko Okamura ,  Hiroshi Tone ,  Rokuro Okamoto

发明人： Akira Arisawa ,  Naoto Kawamura ,  Ikuo Kojima ,  Yasushi Okumura ,  Kazuhiko Okamura ,  Hiroshi Tone ,  Rokuro Okamoto

IPC分类号： C07H17/08 ,  C12N1/15 ,  C12N9/00 ,  C12N9/10 ,  C12N15/00 ,  C12N15/09 ,  C12N15/31 ,  C12P19/62 ,  C12R1/19 ,  C12R1/465 ,  C12R1/54

CPC分类号： C07H17/08 ,  C12N9/1029 ,  C12P19/62

摘要： A DNA fragment containing acyB gene coding for an enzyme capable of acylating the 4"-position of a macrolide antibiotic, said DNA fragment being derived from a microorganism of the genus Streptomyces, having a size of about 3.1 kb and being characterized by the restriction endonuclease map shown in FIG. 1 , and a DNA restriction fragment resulting from digestion with a restriction endonuclease; a recombinant DNA plasmid containing this DNA fragment; a microorganism transformed with this recombinant plasmid; and a process for producing a 4"-acylated macrolide antibiotic by using the transformant.

公开(公告)号：US08008049B2

公开(公告)日：2011-08-30

申请号：US11630689

申请日：2005-07-19

申请人： Kazuhiro Machida ,  Akira Arisawa ,  Susumu Takeda ,  Masashi Yoshida ,  Toshio Tsuchida

发明人： Kazuhiro Machida ,  Akira Arisawa ,  Susumu Takeda ,  Masashi Yoshida ,  Toshio Tsuchida

IPC分类号： C12P17/16 ,  C07H21/04 ,  C07K2/00 ,  C12N9/00 ,  C12N15/63 ,  C12N1/00

CPC分类号： C12P17/08 ,  C07K14/36 ,  C12N9/0071 ,  C12N9/0073 ,  C12N15/52 ,  C12P17/162

摘要： The present invention provides polypeptides that participate in the biosynthesis of the pladienolide macrolide compounds, DNA that encodes these polypeptides and variants of this DNA, transformants that maintain all or a portion of this DNA or variant thereof, and a method of producing the pladienolide macrolide compounds using these transformants. More particularly, it provides an isolated pure DNA that contains at least one region encoding a polypeptide that participates in pladienolide biosynthesis; polypeptide encoded by this DNA; a self-replicating or integrated-replicating recombinant plasmid carrying this DNA; a transformant maintaining this DNA; and a method of producing a pladienolide, characterized by culturing this transformant on culture medium and collecting pladienolide from this culture medium.

摘要翻译： 本发明提供了参与扁平二烯内酰脲大环内酯化合物的生物合成的多肽，编码这些多肽的DNA和该DNA的变体，保持该DNA或其变体的全部或一部分的转化体，以及生产该扁桃苷内酰胺大环内酯化合物的方法 使用这些转化体。 更具体地，其提供了分离的纯DNA，其含有至少一个编码参与泛二烯内酯生物合成的多肽的区域; 由该DNA编码的多肽; 携带该DNA的自我复制或整合复制重组质粒; 维持该DNA的转化体 以及生产普拉二烯内酯的方法，其特征在于在培养基上培养该转化体并从该培养基中收集普拉二烯内酯。

公开(公告)号：US20060234337A1

公开(公告)日：2006-10-19

申请号：US10510826

申请日：2003-04-11

申请人： Akira Arisawa ,  Ayako Kumeda

发明人： Akira Arisawa ,  Ayako Kumeda

IPC分类号： C12P21/06 ,  C12N9/02 ,  C07H21/04 ,  C12N15/74 ,  C12N1/21

CPC分类号： C12N15/70 ,  C12N9/0077 ,  C12N9/0095

摘要： This invention relates to a system for the expression of cytochrome P-450 gene in host Escherichia coli, and provides Escherichia coli which contains actinomycete ferredoxin gene and also ferredoxin gene and ferredoxin reductase gene which are xenogenic to Escherichia coli. Thus, this invention is useful for the promotion of effective single oxygen atom insertional reaction of a substrate organic compound.

摘要翻译： 本发明涉及用于在宿主大肠杆菌中表达细胞色素P-450基因的系统，并提供含有放线菌铁氧还蛋白基因的大肠杆菌以及对大肠杆菌异源的铁氧还蛋白基因和铁氧还蛋白还原酶基因。 因此，本发明可用于促进底物有机化合物的有效单氧原子插入反应。

公开(公告)号：US06361987B1

公开(公告)日：2002-03-26

申请号：US09514340

申请日：2000-02-28

申请人： Akira Arisawa ,  Motoko Matsufuji ,  Takurou Tsuruta ,  Kazuyuki Dobashi ,  Takashi Nakashima ,  Kunio Isshiki ,  Takeo Yoshioka

发明人： Akira Arisawa ,  Motoko Matsufuji ,  Takurou Tsuruta ,  Kazuyuki Dobashi ,  Takashi Nakashima ,  Kunio Isshiki ,  Takeo Yoshioka

IPC分类号： C12N914

CPC分类号： C07D211/88 ,  C07D211/90 ,  C12N9/18 ,  C12N9/78 ,  C12P17/12 ,  C12P41/006

摘要： This invention provides DNA fragments for efficiently preparing a protein having asymmetric hydrolase activity for 4-substituted 1,4-dihydropyridine derivatives, transformants obtained by using such a DNA fragment, and a process for preparing the aforesaid enzyme. Specifically, an isolated gene derived from the chromosome of Streptomyces viridosporus and encoding a protein having asymmetric hydrolase activity for 4-substituted 1,4-dihydropyridine derivatives, plasmids having the aforesaid gene integrated thereinto, transformants obtained by using such a plasmid, and a process for preparing the aforesaid enzyme by using such a transformant are disclosed. A process for the enzymatic conversion of 4-substituted 1,4-dihydropyridine derivatives by using the aforesaid enzyme is also disclosed.

摘要翻译： 本发明提供了用于有效制备对4-取代的1,4-二氢吡啶衍生物具有不对称水解酶活性的蛋白质的DNA片段，以及通过使用这样的DNA片段得到的转化体及其制备方法。 具体来说，来源于链球菌链球菌的染色体的分离的基因，编码对4-取代的1,4-二氢吡啶衍生物具有不对称的水解酶活性的蛋白质，将上述基因整合到其中的质粒，使用这样的质粒获得的转化体， 公开了通过使用这种转化体制备上述酶的方法。 还公开了通过使用上述酶酶促转化4-取代的1,4-二氢吡啶衍生物的方法。

公开(公告)号：US5322937A

公开(公告)日：1994-06-21

申请号：US708866

申请日：1991-05-31

申请人： Akira Arisawa ,  Naoto Kawamura ,  Ikuo Kojima ,  Kazuhiko Okamura ,  Hiroshi Tone ,  Rokuro Okamoto

发明人： Akira Arisawa ,  Naoto Kawamura ,  Ikuo Kojima ,  Kazuhiko Okamura ,  Hiroshi Tone ,  Rokuro Okamoto

IPC分类号： C07H17/08 ,  C12N1/21 ,  C12N9/10 ,  C12N15/55 ,  C12N15/76 ,  C12P19/62 ,  C12P21/06 ,  A01N37/18 ,  C07H19/00

CPC分类号： C07H17/08 ,  C12N15/76 ,  C12N9/1029 ,  C12P19/62 ,  Y10S435/822

摘要： A DNA fragment or a restriction enzyme-digested DNA fragment, the DNA fragment containing a gene, acyA, encoding a 3-acylation enzyme for macrolide antibiotics, characterized by being derived from a strain belonging to the genus Streptomyces, having a size of about 1.8 kb or about 3.2 kb, and having a DNA base sequence shown in a restriction enzyme map shown in FIG. 1(A) or FIG. 1(B), respectively, in the attached drawing. 3-Acylated macrolide antibiotics can be produced advantageously using macrolide antibiotics-producing bacteria transformed with vector plasmids having inserted therein the DNA fragment.

摘要翻译： DNA片段或限制酶消化的DNA片段，含有编码大环内酯类抗生素3-酰化酶的基因acyA的DNA片段，其特征在于衍生自属于链霉菌属（Streptomyces）属的菌株，大小约为1.8 kb或约3.2kb，并且具有如图1所示的限制酶图谱所示的DNA碱基序列。 图1（A） 1（B），附图中。 3-酰基化大环内酯类抗生素可以有利地用大环内酯抗生素产生细菌转化，其中插入有DNA片段的载体质粒转化。