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1. 发明申请

US20060148039A1 Methylotroph producing mammalian type sugar chain 有权
标题翻译：生产甲基鱼油的哺乳动物类型的糖链
公开(公告)号：US20060148039A1
公开(公告)日：2006-07-06
申请号：US10511436
申请日：2003-04-28
申请人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasumori Chiba
发明人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasumori Chiba
IPC分类号： C12P19/28 , C12N15/74 , C12N1/18
CPC分类号： C12P19/26 , C07K16/243 , C07K2317/21 , C12N1/14 , C12N9/0006 , C12N9/0008 , C12N9/1048 , C12N9/1051 , C12N9/2488 , C12N9/60 , C12N9/88 , C12N9/93 , C12P21/02 , C12Y101/01085 , C12Y101/03013 , C12Y102/01012 , C12Y302/01024 , C12Y402/01019 , C12Y603/02006
摘要： This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.
摘要翻译：本发明提供一种生产包含哺乳动物型糖链的糖蛋白的方法，其特征在于该方法包括将α-1,2-甘露糖苷酶基因导入具有糖链生物合成酶基因突变的甲基营养酵母中，因此 α-1,2-甘露糖苷酶基因在酵母中有效启动子的控制下表达; 在介质中培养具有转移到其中的异源基因的甲基营养酵母细胞; 从培养物中获得包含哺乳动物型糖链的糖蛋白。使用新制造的具有糖链突变的甲基营养酵母，可以大量生产与由哺乳动物细胞如人细胞产生的高甘露糖型糖链或包含这种中性糖链的糖蛋白相同的中性糖链纯度高。通过将哺乳动物型糖链生物合成基因导入到上述突变体中，可以有效地制备哺乳动物型糖链，例如杂合体或复合物，或包含哺乳动物型糖链的蛋白质。

2. 发明授权

US07972809B2 Methylotrophic yeast producing mammalian type sugar chain 有权
标题翻译：甲基营养酵母产生哺乳动物型糖链
公开(公告)号：US07972809B2
公开(公告)日：2011-07-05
申请号：US10511436
申请日：2003-04-28
申请人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasunori Chiba
发明人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasunori Chiba
IPC分类号： C12P21/00 , C12N15/01
CPC分类号： C12P19/26 , C07K16/243 , C07K2317/21 , C12N1/14 , C12N9/0006 , C12N9/0008 , C12N9/1048 , C12N9/1051 , C12N9/2488 , C12N9/60 , C12N9/88 , C12N9/93 , C12P21/02 , C12Y101/01085 , C12Y101/03013 , C12Y102/01012 , C12Y302/01024 , C12Y402/01019 , C12Y603/02006
摘要： This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.
摘要翻译：本发明提供一种生产包含哺乳动物型糖链的糖蛋白的方法，其特征在于该方法包括将α-1,2-甘露糖苷酶基因导入具有糖链生物合成酶基因突变的甲基营养酵母中，因此 α-1,2-甘露糖苷酶基因在酵母中有效启动子的控制下表达; 在介质中培养具有转移到其中的异源基因的甲基营养酵母细胞; 从培养物中获得包含哺乳动物型糖链的糖蛋白。使用新制造的具有糖链突变的甲基营养酵母，可以大量生产与由哺乳动物细胞如人细胞产生的高甘露糖型糖链或包含这种中性糖链的糖蛋白相同的中性糖链纯度高。通过将哺乳动物型糖链生物合成基因导入到上述突变体中，可以有效地制备哺乳动物型糖链，例如杂合体或复合物，或包含哺乳动物型糖链的蛋白质。

3. 发明申请

US20070087426A1 Novel transferase and amylase, process for producing the enzymes, use thereof, and gene coding for the same 审中-公开
标题翻译：新型转移酶和淀粉酶，用于生产酶的方法，其用途及其编码基因
公开(公告)号：US20070087426A1
公开(公告)日：2007-04-19
申请号：US11526632
申请日：2006-09-26
申请人： Masaru Kato , Yutaka Miura , Masako Kettoku , Akihiro Iwamatsu , Kazuo Kobayashi , Toshihiro Komeda
发明人： Masaru Kato , Yutaka Miura , Masako Kettoku , Akihiro Iwamatsu , Kazuo Kobayashi , Toshihiro Komeda
IPC分类号： C12N9/10 , C07H21/04
CPC分类号： C12N9/2411 , C12N9/1048 , C12N9/2408 , C12N9/2414 , C12N9/2417 , C12P19/14
摘要： The invention provides a novel transferase that acts on a saccharide, as a substrate, composed of at least three sugar units wherein at least three glucose residues on the reducing end are linked α-1,4 so as to transfer the α-1,4 lingages to a α-1, α-1 linkages; a process for producing the transferase; a gene coding for the same; and a process for producing an oligosaccharide by using the same. Also provided are a novel amylase that has a principal activity of acting on a saccharide, as a substrate, composed of at least three sugar units wherein at least three sugar units on the reducing end side are glucose units and the linkage between the first and the second glucose units is α-1, α-1 while the linkage between the second and the third glucose units is α-1,4 so as to liberate α, α-trehalose by hydrolyzing the α-1,4 linkage and another activity of hydrolyzing the α-1,4 linkage within the molecular chain of the substrate and that liberates disaccharides and/or monosaccharides as the principal final products; a process for producing the amylase; a gene coding for the same; and a process for producing α, α-trehalose by using a combination of the transferase and the amylase.
摘要翻译：本发明提供了一种新的转移酶，其作用为由至少三个糖单元组成的作为底物的糖，其中还原端上的至少三个葡萄糖残基与α-1,4连接以转移α-1,4 属于α-1，α-1键; 生产转移酶的方法; 编码相同的基因; 以及使用该寡糖的方法。还提供了一种新的淀粉酶，其具有作用于由至少三个糖单元组成的糖类作为底物的主要活性，其中还原端侧的至少三个糖单元是葡萄糖单元，第一和第第二葡萄糖单位是α-1，α-1，而第二和第三葡萄糖单元之间的连接是α-1,4，以便通过水解α-1,4键和另一种活性的α-1,4键来释放α，α-海藻糖水解基质分子链内的α-1,4键，释放出二糖和/或单糖作为主要的最终产物; 淀粉酶的制造方法; 编码相同的基因; 以及通过使用转移酶和淀粉酶的组合产生α，α-海藻糖的方法。

4. 发明授权

US06423510B1 Candida boidini dihydroxyacetone synthase promoter 失效
标题翻译：念珠菌boidini二羟基丙酮合酶启动子
公开(公告)号：US06423510B1
公开(公告)日：2002-07-23
申请号：US09622540
申请日：2000-08-18
申请人： Toshihiro Komeda , Keiji Kondo , Nobuo Kato , Yasuyoshi Sakai
发明人： Toshihiro Komeda , Keiji Kondo , Nobuo Kato , Yasuyoshi Sakai
IPC分类号： C12P2106
CPC分类号： C12N15/815 , C12N9/1022
摘要： This invention relates to a promoter for dihydroxyacetone synthase gene from Candida boidinii; an expression cassette or vector comprising the promoter and a heterologous gene; a transformant transformed with this vector; and a method for producing an expression product of the heterologous gene which comprises culturing the transformant to express the heterologous gene.
摘要翻译：本发明涉及来自布氏念珠菌的二羟基丙酮合成酶基因的启动子; 包含启动子和异源基因的表达盒或载体; 用该载体转化的转化体; 以及用于产生异源基因的表达产物的方法，其包括培养转化体以表达异源基因。

5. 发明授权

US06274340B1 DNA sequences enhancing promoter activity 有权
标题翻译： DNA序列增强启动子活性
公开(公告)号：US06274340B1
公开(公告)日：2001-08-14
申请号：US09297053
申请日：1999-04-28
申请人： Toshihiro Komeda
发明人： Toshihiro Komeda
IPC分类号： C12P2106
CPC分类号： C12N15/85 , C12N15/80 , C12N2830/00 , C12N2830/15 , C12N2830/702
摘要： The present invention relates to a DNA comprising the nucleotide sequence shown in SEQ ID NO:1, or a DNA for enhancing promoter activity which comprises a nucleotide sequence having deletions, substitutions, additions or insertions of one or more nucleotides in the nucleotide sequence shown in SEQ ID NO:1; to a mutant promoter comprising one or more fragments of the DNA; to a recombinant expression vector comprising the mutant promoter together with a heterologous gene; to a transformant prepared by transforming a host cell with the vector; to a process for preparing an expression product, comprising culturing the transformant in a medium, and recovering the expression product of a heterologous gene from the obtained culture; and to a method for enhancing a promoter activity, characterized in that one or more fragments of the above-defined DNA are located at at least one position preceding, following or within a selective promoter in a forward or reverse direction.
摘要翻译：本发明涉及包含SEQ ID NO：1所示核苷酸序列的DNA或用于增强启动子活性的DNA，其包含在SEQ ID NO：1所示的核苷酸序列中具有一个或多个核苷酸的缺失，替换，添加或插入的核苷酸序列 SEQ ID NO：1; 涉及包含DNA的一个或多个片段的突变启动子; 涉及包含突变启动子与异源基因的重组表达载体; 通过用载体转化宿主细胞制备的转化体; 涉及一种制备表达产物的方法，包括在培养基中培养转化体，并从获得的培养物中回收异源基因的表达产物; 以及用于增强启动子活性的方法，其特征在于，上述定义的DNA的一个或多个片段位于选择性启动子前后的至少一个位置，在前向或反向方向。

6. 发明申请

US20080096266A1 Method for Stabilizing the Antiallergic Activity of Lactic Acid Bacteria Against High-Temperature Treatment, Stabilized Compositons, and Food and Drink 审中-公开
标题翻译：稳定乳酸菌抗高温处理，稳定化合物和食品饮料的抗过敏活性的方法
公开(公告)号：US20080096266A1
公开(公告)日：2008-04-24
申请号：US11662077
申请日：2005-09-08
申请人： Toshihiro Komeda , Kumiko Ikado , Koichiro Yamauchi , Akira Saiki , Kiyoko Sadakari
发明人： Toshihiro Komeda , Kumiko Ikado , Koichiro Yamauchi , Akira Saiki , Kiyoko Sadakari
IPC分类号： C12N1/20 , A23L1/30 , A23L2/52 , A61P37/08
CPC分类号： C12N1/20 , A23L2/52 , A23L33/135 , A23V2002/00 , A23Y2220/67 , A23Y2240/65 , A23V2200/10 , A23V2250/214 , A23V2250/2132 , A23V2200/3204
摘要： The present invention provides a method for stabilizing an antiallergic activity of lactic acid bacteria against a high-temperature treatment and a composition for the stabilization, which can maintain an antiallergic activity of lactic acid bacteria against a high-temperature treatment, and foods and drinks such as drinks having a stabilized antiallergic activity of lactic acid bacteria, and a method for producing the same. The antiallergic activity of lactic acid bacteria can be maintained stably by making the lactic acid bacteria and polyphenols coexist, even when high-temperature treatments are conducted. As polyphenols used in the present invention, polyphenols extracted from a plant such as green tea-derived polyphenols can be used. In the present invention, various lactic acid bacteria having an antiallergic activity can be used, and in particular, lactic acid bacteria having a high antiallergic activity can be advantageously used as such lactic acid bacteria. The method of the present invention can be applied to foods and drinks for which high-temperature heating treatments are conducted in the process of their production, distribution, or when they are taken.
摘要翻译：本发明提供了一种用于稳定乳酸菌抗高温处理的抗过敏活性的方法和用于稳定化的组合物，其可以保持乳酸菌耐高温处理的抗过敏活性，以及食品和饮料作为乳酸菌具有稳定的抗过敏活性的饮料及其制造方法。即使进行高温处理也能够使乳酸菌和多酚共存，可以稳定地维持乳酸菌的抗过敏活性。作为本发明中使用的多酚，可以使用从植物提取的多酚，例如绿茶衍生的多酚。在本发明中，可以使用具有抗过敏活性的各种乳酸菌，特别是可以有利地使用具有高抗过敏活性的乳酸菌作为乳酸菌。本发明的方法可以应用于在其生产，分配或摄取过程中进行高温加热处理的食品和饮料。

7. 发明授权

US06001590A Promoter and terminator sequences of formate dehydrogenase gene of Candida boidinii 失效
标题翻译：博伊丁假丝酵母的甲酸脱氢酶基因的启动子和终止子序列
公开(公告)号：US06001590A
公开(公告)日：1999-12-14
申请号：US817926
申请日：1997-05-09
申请人： Toshihiro Komeda , Hisako Suda , Yukio Tamai , Akihiro Iwamatsu , Nobuo Kato , Yasuyoshi Sakai
发明人： Toshihiro Komeda , Hisako Suda , Yukio Tamai , Akihiro Iwamatsu , Nobuo Kato , Yasuyoshi Sakai
IPC分类号： C12N1/15 , C12N9/02 , C12N15/81 , C12N15/31 , C12P21/00
CPC分类号： C12N9/0008 , C12N15/815
摘要： A promoter for a formate dehydrogenase gene from Candida boidinii, substantially comprising a 190 bp or more continuous nucleotide sequence selected from the nucleotide sequence of SEQ ID NO:1; a promoter for a formate dehydrogenase gene from Candida boidinii, substantially comprising the nucleotide sequence of SEQ ID NO:1, 48, 49 or 50; a terminator for a formate dehydrogenase gene from Candida boidinii, substantially comprising the nucleotide sequence of SEQ ID NO:2; a gene expression cassette comprising said promoter, a heterologous gene and said terminator; a recombinant expression vector comprising said gene expression cassette; a transformant transformed with said recombinant expression vector; a process for producing an expression product of a heterologous gene, which comprises culturing said transformant and recovering an expression product of a heterologous gene from the culture.
摘要翻译： PCT No.PCT / JP96 / 02597 371日期：1997年5月9日 102（e）日期1997年5月9日PCT提交1996年9月12日PCT公布。公开号WO97 / 10345 日期：1997年3月20日来自布氏念珠菌的甲酸脱氢酶基因的启动子，基本上包含选自SEQ ID NO：1的核苷酸序列的190bp以上的连续核苷酸序列; 来自布氏念珠菌的甲酸脱氢酶基因的启动子，基本上包含SEQ ID NO：1,49,49或50的核苷酸序列; 来自布氏念珠菌的甲酸脱氢酶基因的终止子，基本上包含SEQ ID NO：2的核苷酸序列; 包含所述启动子，异源基因和所述终止子的基因表达盒; 包含所述基因表达盒的重组表达载体; 用所述重组表达载体转化的转化体; 用于产生异源基因的表达产物的方法，其包括培养所述转化体并从培养物中回收异源基因的表达产物。

8. 发明申请

US20060088513A1 Antiallergic composition 审中-公开
标题翻译：抗过敏组合物
公开(公告)号：US20060088513A1
公开(公告)日：2006-04-27
申请号：US10504901
申请日：2004-02-27
申请人： Sayo Inoue , Toshio Fujii , Daisuke Fujiwara , Akira Saiki , Minoru Takahashi , Koichiro Yamauchi , Masami Gotou , Satoshi Nishida , Keiji Deuchi , Hideyuki Wakabayashi , Toshihiro Komeda
发明人： Sayo Inoue , Toshio Fujii , Daisuke Fujiwara , Akira Saiki , Minoru Takahashi , Koichiro Yamauchi , Masami Gotou , Satoshi Nishida , Keiji Deuchi , Hideyuki Wakabayashi , Toshihiro Komeda
IPC分类号： A61K35/74 , C12N1/20
CPC分类号： A23C9/1234 , A23F3/166 , A23F3/34 , A23F5/246 , A23L2/52 , A23L33/135 , A23Y2220/67 , A61K35/747
摘要： The object of the present invention is to provide an antiallergic composition useful to prevention/treatment of allergy, its preparation method, and its use as foods or drinks or the like. As for means for solving the object, it is to provide an antiallergic composition comprising as active ingredients the lactic acid bacteria showing equal interleukin 12 production level compared to when Lactobacillus paracasei KW 3110 strain is used as the lactic acid bacteria to be tested, or showing 60% or more of interleukin 12 production level compared to when Lactobacillus paracasei KW 3110 strain is used, and showing less than 50% of interleukin 4 production level of a control wherein the lactic acid bacteria are not added, in case lymphocytes derived from mouse spleen sensitized with ovalbumin are suspended in a medium containing ovalbumin, and cultured by adding the lactic acid bacteria to be tested. The present invention encompasses a method for obtaining the lactic acid bacteria with antiallergic activity being the active ingredients of the antiallergic composition of the present invention, and the use of the antiallergic composition of the present invention to foods and beverages and the like, and further by formulation, the use as antiallergic agent administered orally.
摘要翻译：本发明的目的是提供一种用于预防/治疗过敏的抗过敏组合物，其制备方法及其作为食品或饮料等的用途。作为解决目的的方法，提供一种抗过敏组合物，其与作为待测试乳酸菌的副乳酸杆菌KW3110株相比，含有作为活性成分的乳酸菌显示出相等的白细胞介素12的产生水平，或显示出与使用副干酪乳杆菌KW3110菌株相比，白细胞介素12的产生水平高达60％以上，并且，如果不添加乳酸菌，则显示少于50％的白细胞介素4生产水平，如果来自小鼠脾的淋巴细胞用卵白蛋白敏化的物质悬浮在含有卵清蛋白的培养基中，并通过加入待测试的乳酸菌进行培养。本发明包括获得具有抗过敏活性的乳酸菌的方法，其为本发明的抗过敏组合物的活性成分，以及本发明的抗过敏组合物在食品和饮料等中的用途，进一步由制剂，口服给药作为抗过敏剂。

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