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1. 发明授权

US07285404B1 Cyclic depsipeptide synthetase and method for recombinant production 失效
标题翻译：循环depipipeptide合成酶和重组生产方法
公开(公告)号：US07285404B1
公开(公告)日：2007-10-23
申请号：US10070387
申请日：2000-09-07
申请人： Naoki Midoh , Kaoru Okakura , Koichi Miyamoto , Manabu Watanabe , Koji Yanai , Tetsuya Yasutake , Sato Aihara , Takafumi Futamura , Horst Kleinkauf , Takeshi Murakami
发明人： Naoki Midoh , Kaoru Okakura , Koichi Miyamoto , Manabu Watanabe , Koji Yanai , Tetsuya Yasutake , Sato Aihara , Takafumi Futamura , Horst Kleinkauf , Takeshi Murakami
IPC分类号： C12N9/00
CPC分类号： C12N9/00 , C12P21/00
摘要： The present invention provides an enzyme that synthesizes a cyclic depsipeptide, particularly the substance 1022, and a gene thereof. A cyclic depsipeptide synthetase according to the present invention comprises (a) an amino acid sequence of SEQ ID NO: 2 or (b) a modified amino acid sequence of the amino acid sequence of SEQ ID NO: 2 that have one or more modifications selected from a substitution, a deletion, an addition and an insertion and has cyclic depsipeptide synthetase activity. A cyclic depsipeptide synthetase gene according to the present invention comprises a nucleotide sequence encoding a cyclic depsipeptide synthetase. The present invention also provides a recombinant vector and a transformant for expressing the cyclic depsipeptide synthetase, and a mass production system for the cyclic depsipeptide. The present invention further provides a method for producing the cyclic depsipeptide synthetase.
摘要翻译：本发明提供合成环状缩肽，特别是物质1022及其基因的酶。根据本发明的循环倍肽合成酶包含（a）SEQ ID NO：2的氨基酸序列或（b）SEQ ID NO：2的氨基酸序列的修饰的氨基酸序列，其具有一个或多个修饰选择从取代，缺失，添加和插入，并具有循环衰老肽合成酶活性。根据本发明的环状缩肽酶合成酶基因包含编码环状倍肽合成酶的核苷酸序列。本发明还提供了用于表达环状缩酚酸肽合成酶的重组载体和转化体，以及用于环状缩酚酸肽的批量生产系统。本发明还提供了一种用于制备环状缩酚酸肽合成酶的方法。

2. 发明授权

US07109018B1 Transformant producing secondary metabolite modified with functional group and novel biosynthesis genes 失效
标题翻译：用功能组和新生物合成基因修饰的转化体产生次级代谢产物
公开(公告)号：US07109018B1
公开(公告)日：2006-09-19
申请号：US10089514
申请日：2000-09-29
申请人： Koji Yanai , Kaoru Okakura , Shohei Yasuda , Manabu Watanabe , Koichi Miyamoto , Naoki Midoh , Takeshi Murakami
发明人： Koji Yanai , Kaoru Okakura , Shohei Yasuda , Manabu Watanabe , Koichi Miyamoto , Naoki Midoh , Takeshi Murakami
IPC分类号： C12N1/20 , C12N5/10 , C07H21/04
CPC分类号： C12N9/0004 , C12N9/88 , C12N9/90 , C12N15/52 , C12P13/001 , C12P13/02
摘要： An objective of the present invention is to provide a transformant altered so as to produce a secondary metabolite in which a benzene ring of the secondary product is modified at the para-position with a functional group containing a nitrogen atom. A transformant according to the present invention is a transformant of an organism producing a secondary metabolite having a benzene ring skeleton without substitution at the para-position with a functional group containing a nitrogen atom, said transformant being transformed by introducing genes involved in a biosynthetic pathway from chorismic acid to p-aminophenylpyruvic acid, including a gene encoding an amino acid sequence (SEQ ID NO: 2) having 4-amino-4-deoxychorismic acid synthase activity, a gene encoding an amino acid sequence (SEQ ID NO: 4) having 4-amino-4-deoxychorismic acid mutase activity and a gene encoding an amino acid sequence (SEQ ID NO: 6) having 4-amino-4-deoxyprephenic acid dehydrogenase activity, so as to produce a secondary metabolite having a benzene ring skeleton substituted at the para-position with a functional group containing a nitrogen atom. Another objective of the present invention is to provide a novel gene involved in the biosynthetic pathway from chorismic acid to p-aminophenylpyruvic acid. A novel gene according to the present invention comprises genes encoding the amino acid sequences of SEQ ID NOs: 2, 4 and 6 or modified sequences thereof.
摘要翻译：本发明的目的是提供改性的转化体，以便产生次级代谢物，其中次级产物的苯环在对位上用含氮原子的官能团改性。根据本发明的转化体是产生具有苯环骨架的次级代谢物的生物体的转化体，其在具有含氮原子的官能团的对位没有取代，所述转化体通过引入参与生物合成途径的基因而转化包括编码氨基酸序列（SEQ ID NO：4）的氨基酸序列（SEQ ID NO：2）的编码氨基酸序列（SEQ ID NO：4）的氨基酸序列（SEQ ID NO：2）的基因的氨基苯基丙酮酸，具有4-氨基-4-脱氧酸性变位酶活性的基因，以及编码具有4-氨基-4-脱氧蝶呤酸脱氢酶活性的氨基酸序列（SEQ ID NO：6）的基因，以产生具有苯环骨架的次级代谢物在对位上用含有氮原子的官能团取代。本发明的另一个目的是提供一种涉及从氯酸到对氨基苯基丙酮酸生物合成途径的新基因。根据本发明的新基因包含编码SEQ ID NO：2,4和6的氨基酸序列的基因或其修饰序列。

3. 发明授权

US07432102B2 Transformants producing substance pf1022 derivatives, methods for producing the same, and novel biosynthesis genes 失效
标题翻译：产生物质pf1022衍生物的转化体，其制备方法和新的生物合成基因
公开(公告)号：US07432102B2
公开(公告)日：2008-10-07
申请号：US10472587
申请日：2002-03-22
申请人： Koji Yanai , Naomi Sumida , Manabu Watanabe , Tatsuki Moriya , Takeshi Murakami
发明人： Koji Yanai , Naomi Sumida , Manabu Watanabe , Tatsuki Moriya , Takeshi Murakami
IPC分类号： C12N1/00 , C12P1/00 , C12P17/00 , C12P15/00 , C12P13/00 , C12P7/00 , C12P7/24 , C12P1/02
CPC分类号： C12P17/14 , C12N15/52
摘要： An objective of the present invention is to provide a method for producing substance PF1022 derivatives, in particular PF1022-220 and PF1022-260, by a direct fermentation method, and a transformant to be used for this method. According to the present invention, there is provided a transformant producing substance FF1022 derivatives, which can be obtained by introducing a genes involved in a biosynthetic pathway from chorismic acid to p-aminophenylpyruvic acid, including a papA gene encoding 4-amino-4-deoxychorismate synthase. which gene comprises the DNA sequence encoding the amino acid sequence of SEQ ID NO: 2; a papB gene encoding 4-amino-4-deoxyclaismate mutase, which gene comprises the DNA sequence encoding the amino acid sequence of SEQ ID NO: 4; and a papC gene encoding 4-amino-4-deoxyprepbenate dehydrogenase, which gene comprises the DNA sequence encoding the amino acid sequence of SEQ ID NO: 6, into a phenylalanine auxotrophic host induced from an organism that produces a substance PF1022. According to the present invention, there is also provided a process of producing substance PF1022 derivative, comprising steps of culturing the above-mentioned transformant and collecting the substance PF1022 derivatives.
摘要翻译：本发明的目的是提供通过直接发酵方法生产物质PF1022衍生物，特别是PF1022-220和PF1022-260的方法，以及用于该方法的转化体。根据本发明，提供了转化体生产物质FF1022衍生物，其可以通过将参与生物合成途径的基因从氯磺酸引入对氨基苯基丙酮酸而获得，包括编码4-氨基-4-脱氧胸苷酸的papA基因合酶。该基因包含编码SEQ ID NO：2的氨基酸序列的DNA序列; 编码4-氨基-4-脱氧胆酸酯变位酶的papB基因，该基因包含编码SEQ ID NO：4的氨基酸序列的DNA序列; 和编码4-氨基-4-脱氧降磷酸脱氢酶的papC基因，该基因包含编码SEQ ID NO：6的氨基酸序列的DNA序列，由产生PF1022的生物体诱导的苯丙氨酸营养缺陷型宿主中。根据本发明，还提供了生产物质PF1022衍生物的方法，包括培养上述转化体并收集物质PF1022衍生物的步骤。

4. 发明申请

US20070099265A1 Surfactant tolerant cellulase and method for modification thereof 有权
标题翻译：表面活性剂耐受性纤维素酶及其修饰方法
公开(公告)号：US20070099265A1
公开(公告)日：2007-05-03
申请号：US10582002
申请日：2004-12-07
申请人： Manabu Watanabe , Koji Yanai , Yumiko Tsuyuki
发明人： Manabu Watanabe , Koji Yanai , Yumiko Tsuyuki
IPC分类号： C12P21/06 , C12N9/42 , C07H21/04 , C12N15/74 , C12N1/16
CPC分类号： C12N9/2437 , C12Y302/01004
摘要： A method for suppressing a reduction in an endoglucanase activity in the presence of a surfactant, characterized by modifying a protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid, to a protein having the N-terminus of pyroglutamic acid, is disclosed. Further, a modified protein having an endoglucanase activity wherein the N-terminal amino acid is converted into pyroglutamic acid by an amino acid modification, a polynucleotide encoding the protein, an expression vector comprising the polynucleotide, a host cell transformed with the expression vector, and a process for producing the protein by cultivating the host cell, are disclosed.
摘要翻译：一种抑制表面活性剂存在下内切葡聚糖酶活性降低的方法，其特征在于将具有N-端是除焦谷氨酸以外的氨基酸的具有内切葡聚糖酶活性的蛋白质修饰为具有N末端的蛋白质焦谷氨酸。此外，具有内切葡聚糖酶活性的修饰蛋白质，其中N-末端氨基酸通过氨基酸修饰转化为焦谷氨酸，编码该蛋白质的多核苷酸，包含该多核苷酸的表达载体，用表达载体转化的宿主细胞，以及公开了通过培养宿主细胞产生蛋白质的方法。

5. 发明授权

US08569033B2 Surfactant tolerant cellulase and method for modification thereof 有权
标题翻译：表面活性剂耐受性纤维素酶及其修饰方法
公开(公告)号：US08569033B2
公开(公告)日：2013-10-29
申请号：US10582002
申请日：2004-12-07
申请人： Manabu Watanabe , Koji Yanai , Yumiko Tsuyuki
发明人： Manabu Watanabe , Koji Yanai , Yumiko Tsuyuki
IPC分类号： C12N9/42 , C12N9/26 , C12N1/00 , C12N15/00 , C12P21/06 , C07H21/04
CPC分类号： C12N9/2437 , C12Y302/01004
摘要： A method for suppressing a reduction in an endoglucanase activity in the presence of a surfactant, characterized by modifying a protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid, to a protein having the N-terminus of pyroglutamic acid, is disclosed. Further, a modified protein having an endoglucanase activity wherein the N-terminal amino acid is converted into pyroglutamic acid by an amino acid modification, a polynucleotide encoding the protein, an expression vector comprising the polynucleotide, a host cell transformed with the expression vector, and a process for producing the protein by cultivating the host cell, are disclosed.
摘要翻译：一种抑制表面活性剂存在下内切葡聚糖酶活性降低的方法，其特征在于将具有N-端是除焦谷氨酸以外的氨基酸的具有内切葡聚糖酶活性的蛋白质修饰为具有N末端的蛋白质焦谷氨酸。此外，具有内切葡聚糖酶活性的修饰蛋白质，其中N-末端氨基酸通过氨基酸修饰转化为焦谷氨酸，编码该蛋白质的多核苷酸，包含该多核苷酸的表达载体，用表达载体转化的宿主细胞，以及公开了通过培养宿主细胞产生蛋白质的方法。

6. 发明授权

US08383392B2 Transformant and method for production of non-natural antibiotic 有权
标题翻译：用于生产非天然抗生素的转化体和方法
公开(公告)号：US08383392B2
公开(公告)日：2013-02-26
申请号：US12678101
申请日：2008-09-12
申请人： Naomi Sumida , Manabu Watanabe , Koji Yanai
发明人： Naomi Sumida , Manabu Watanabe , Koji Yanai
IPC分类号： C12N1/15 , C12P29/00
CPC分类号： C12P19/56 , C12N9/0006 , C12P17/06 , C12P19/44
摘要： Disclosed is a transformant prepared by introducing a ketoreductase gene involved in the biosynthesis of L-epivancosamine into an actinobacterium originally capable of producing daunorubicin. Also disclosed is a process of efficiently producing a non-natural daunorubicin derivative using the transformant. The transformant is capable of efficiently producing a non-natural daunorubicin derivative such as epidaunorubicin.
摘要翻译：公开了通过将参与L-赖氨替康的生物合成的酮还原酶基因导入本来能够产生柔红霉素的放线杆菌中制备的转化体。还公开了使用转化体有效生产非天然柔红霉素衍生物的方法。转化体能够有效地产生非天然柔红霉素衍生物如表柔比星。

7. 发明申请

US20100255543A1 METHOD FOR PRODUCTION OF NON-NATURAL ANTIBIOTIC 有权
标题翻译：生产非天然抗生素的方法
公开(公告)号：US20100255543A1
公开(公告)日：2010-10-07
申请号：US12678101
申请日：2008-09-12
申请人： Naomi Sumida , Manabu Watanabe , Koji Yanai
发明人： Naomi Sumida , Manabu Watanabe , Koji Yanai
IPC分类号： C12P19/56 , C12N1/21
CPC分类号： C12P19/56 , C12N9/0006 , C12P17/06 , C12P19/44
摘要： Disclosed is a transformant prepared by introducing a ketoreductase gene involved in the biosynthesis of L-epivancosamine into an actinobacterium originally capable of producing daunorubicin. Also disclosed is a process of efficiently producing a non-natural daunorubicin derivative using the transformant. The transformant is capable of efficiently producing a non-natural daunorubicin derivative such as epidaunorubicin.
摘要翻译：公开了通过将参与L-赖氨替康的生物合成的酮还原酶基因导入本来能够产生柔红霉素的放线杆菌中制备的转化体。还公开了使用转化体有效生产非天然柔红霉素衍生物的方法。转化体能够有效地产生非天然柔红霉素衍生物如表柔比星。

8. 发明授权

US5763221A Transformant producing substance PF1022, and method for transforming microorganism belonging to the class hyphomycetes 失效
标题翻译：转化体生产物质PF1022，以及用于转化属于类菌丝体的微生物的方法
公开(公告)号：US5763221A
公开(公告)日：1998-06-09
申请号：US776816
申请日：1997-02-07
申请人： Kaoru Aoyagi , Manabu Watanabe , Kohji Yanai , Takeshi Murakami
发明人： Kaoru Aoyagi , Manabu Watanabe , Kohji Yanai , Takeshi Murakami
IPC分类号： C07K11/02 , C12N1/15 , C12N9/30 , C12N15/74 , C12N15/80 , C12P21/02 , C07K5/12 , C12N15/09
CPC分类号： C12N9/242 , C07K11/02 , C12N15/74 , C12N15/80 , C12R1/645
摘要： A method for transforming strain PF1022, which produces a cyclic depsipeptide (substance PF1022) and belongs to the order Agonomycetales of the class Hyphomycetes has been established by using a plasmid constructed with a promoter, a terminator, a marker gene and a target gene. When the strain PF1022 was transformed by this method, a gene encoding an enzyme, which is the target gene, was transduced into the host. As a result, the utilization of nutrients by the strain PF1022 was improved and, further, its cyclic depsipeptide productivity was also improved.
摘要翻译： PCT No.PCT / JP96 / 01692 Sec。 371日期1997年2月7日 102（e）1997年2月7日PCT PCT 1996年6月19日PCT公布。公开号WO97 / 00944 日期1997年1月9日已经通过使用由启动子，终止子，标记基因构建的质粒和通过启动子，终止子，标记基因构建的质粒来建立转化菌株PF1022的方法，该方法产生环状缩酚酸肽（物质PF1022）并且属于类型菌纲（Agypycycetales）目标基因。当通过该方法转化菌株PF1022时，将编码作为靶基因的酶的基因转导入宿主体。结果，通过菌株PF1022的营养物质的利用得到改善，此外，其环状肽的生产力也得到提高。

9. 发明授权

US06403362B1 Systems for the mass production of proteins or peptides by microorganisms of the genus humicola 有权
标题翻译：通过腐质霉属微生物大规模生产蛋白质或肽的系统
公开(公告)号：US06403362B1
公开(公告)日：2002-06-11
申请号：US09230225
申请日：1999-03-03
申请人： Tatsuki Moriya , Kouichirou Murashima , Kaoru Aoyagi , Naomi Sumida , Manabu Watanabe , Toru Hamaya , Jinichiro Koga , Toshiaki Kono , Takeshi Murakami
发明人： Tatsuki Moriya , Kouichirou Murashima , Kaoru Aoyagi , Naomi Sumida , Manabu Watanabe , Toru Hamaya , Jinichiro Koga , Toshiaki Kono , Takeshi Murakami
IPC分类号： C12N114
CPC分类号： C12N9/2437 , C12N15/80 , C12Y302/01004 , Y10S530/82 , Y10S530/823
摘要： An expressing system which enables a large amount of production of a protein in Humicola, in particular, Humicola insolens, and particularly host-vector systems and a process for producing a protein using the systems, wherein an expression vector comprising the regulator sequences, i.e., the promoter, the signal sequence, and the terminator, of the cellulase NCE1 gene or NCE2 gene derived from Humicola insolens is constructed and used. The expression vector enables highly efficient production of cellulase NCE4, for example, in Humicola insolens at a rate as high as about 4.5 g or more per one liter of culture.
摘要翻译：一种表达系统，其能够在Humicola中大量生产蛋白质，特别是Humicola insolens，特别是宿主 - 载体系统，以及使用该系统产生蛋白质的方法，其中包含调节序列的表达载体，构建并使用来自Humicola insolens的纤维素酶NCE1基因或NCE2基因的启动子，信号序列和终止子。表达载体能够高效率地生产纤维素酶NCE4，例如在Humicola insolens中以每升培养物高达约4.5g或更高的速率。

10. 发明授权

US06913905B1 Regulatory sequences functioning in filamentous fungi 失效
标题翻译：在丝状真菌中起作用的调节序列
公开(公告)号：US06913905B1
公开(公告)日：2005-07-05
申请号：US10070386
申请日：2000-09-07
申请人： Manabu Watanabe , Takeshi Murakami
发明人： Manabu Watanabe , Takeshi Murakami
IPC分类号： C07K14/37 , C12N1/15 , C12N9/24 , C12N15/80 , C12P21/02 , C12N1/00 , C12N5/10 , C12N15/11 , C12N15/63
CPC分类号： C12Y302/01031 , C07K14/37 , C12N9/2402 , C12N15/80
摘要： The present invention provides a promoter and a terminator which coordinately function with the expression of an endogenous gene in a filamentous fungus that belongs to Agonomycetes, particularly in Mycelia sterilia. A promoter of the present invention comprises the nucleotide sequence of SEQ ID NO: 1 and a homologue thereof. A terminator of the present invention comprises the nucleotide sequence of SEQ ID NO: 2 and a homologue thereof. The present invention further provides an expression vector that highly expresses a protein of interest in a filamentous fungus, a transformed filamentous fungus that highly produces a protein of interest, and a process of producing the protein of interest in the transformed filamentous fungus.
摘要翻译：本发明提供了一种启动子和终止子，其与属于天然生物体的丝状真菌中的内源性基因的表达起协调作用，特别是在菌丝体灭菌中。本发明的启动子包含SEQ ID NO：1的核苷酸序列及其同源物。本发明的终止子包含SEQ ID NO：2的核苷酸序列及其同源物。本发明还提供了在丝状真菌中高度表达目的蛋白的表达载体，高度产生目的蛋白质的转化的丝状真菌，以及在转化的丝状真菌中产生目的蛋白质的方法。

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