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1. 发明授权

US08153413B2 High alkaline protease and use thereof 失效
标题翻译：高碱性蛋白酶及其用途
公开(公告)号：US08153413B2
公开(公告)日：2012-04-10
申请号：US11719713
申请日：2005-11-16
申请人： Jie Lu , Zhi jun Li , Si Hung Vo , Yuji Hatada , Ken Takai , Susumu Ito , Koki Horikoshi
发明人： Jie Lu , Zhi jun Li , Si Hung Vo , Yuji Hatada , Ken Takai , Susumu Ito , Koki Horikoshi
IPC分类号： C12N9/00 , C12N9/92 , C12N1/20 , C12N15/00 , C07H21/04
CPC分类号： C12N9/52
摘要： The invention aims to provide a novel alkaline protease having peculiar properties such as high alkali activity, resistance to surfactants and calcium-dependent thermostability and exhibiting excellent performance in highly alkaline detergents, and a gene coding for the amino acid sequence thereof. There is provided an alkaline protease with such properties that an active pH range is from 5 to 13, an optimum pH is approximately 12.6, an optimum temperature is 70° C., no activity drop by heating is observed up to 65° C. at pH 10 and the optimum temperature and the thermostability are not affected by Ca2+ ions. Specifically, there is provided, for example, an alkaline protease having an amino acid sequence constituting a mature enzyme as represented by SEQ ID NO: 3 or an amino acid sequence resulting from deletion, substitution, situs inversus arrangement, addition or insertion of a part of amino acids thereof, or derived from Alkaliphillus transvaalensis. The protease cleaves 26 peptide bonds among 29 peptide bonds of acidic insulin B-chain.
摘要翻译：本发明的目的在于提供一种具有高碱性，耐表面活性和钙依赖性热稳定性等特性，在高碱性洗涤剂中表现出优异性能的新型碱性蛋白酶及其氨基酸序列的编码基因。提供了具有这样的性质的碱性蛋白酶，其活性pH范围为5至13，最佳pH为约12.6，最适温度为70℃，在65℃下不观察到加热下的活性降低 pH 10，最佳温度和热稳定性不受Ca2 +离子的影响。具体地，例如提供了具有SEQ ID NO：3所示的构成成熟酶的氨基酸序列的氨基酸序列的碱性蛋白酶或由缺失，取代，位置反转排列，添加或插入部分的氨基酸，或衍生自变形白菜（Alkaliphillus transvaalensis）。蛋白酶在酸性胰岛素B链的29个肽键中切割26个肽键。

2. 发明申请

US20110008832A1 METHODS FOR STABLY RETAINING FOREIGN GENES IN CELLS 审中-公开
标题翻译：在细胞中稳定保留外源基因的方法
公开(公告)号：US20110008832A1
公开(公告)日：2011-01-13
申请号：US12452645
申请日：2008-07-09
申请人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
发明人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
IPC分类号： C12P21/00 , C12N15/63 , C12N1/21 , C12N1/16 , C12N5/10
CPC分类号： C12N15/64 , C12N9/93 , C12N15/821
摘要： A method for preparing a protein or peptide encoded by a foreign gene by expressing a foreign gene, which comprises the steps of: preparing a recombinant vector comprising in an expressible state a gene encoding an aminoacyl-tRNA synthetase, in which a desired foreign gene has been inserted in an expressible state; preparing a mutant host cell or the like in which a chromosomal gene encoding an aminoacyl-tRNA synthetase has been knocked out; transforming the mutant host cell with the recombinant vector to obtain a transformant; and culturing the transformant to prepare the protein or peptide encoded by the foreign gene. It becomes possible to provide a novel means for permitting the retention of a recombinant DNA cloning vector without employing an antibiotic and without limiting the composition of the medium; and a method for preparing a protein or peptide encoded by a foreign gene by using the means.
摘要翻译：一种通过表达外源基因来制备由外来基因编码的蛋白质或肽的方法，该方法包括以下步骤：制备包含可表达状态的编码氨基-tRNA合成酶的基因的重组载体，其中期望的外源基因具有被插入可表达的状态; 制备其中编码氨基-tRNA合成酶的染色体基因已经被敲除的突变宿主细胞等; 用重组载体转化突变宿主细胞以获得转化体; 并培养转化体以制备由外源基因编码的蛋白质或肽。可以提供一种用于在不使用抗生素的情况下保留重组DNA克隆载体并且不限制培养基的组成的新方法; 以及通过使用该方法制备由外源基因编码的蛋白质或肽的方法。

3. 发明申请

US20080050807A1 Gene encoding alkaline liquefying alpha-amylase 审中-公开
标题翻译：基因编码碱液化α-淀粉酶
公开(公告)号：US20080050807A1
公开(公告)日：2008-02-28
申请号：US10829331
申请日：2004-04-22
申请人： Yuji Hatada , Katsuya Ozaki , Katsutoshi Ara , Shuji Kawai , Susumu Ito
发明人： Yuji Hatada , Katsuya Ozaki , Katsutoshi Ara , Shuji Kawai , Susumu Ito
IPC分类号： C12N15/00 , C07H21/04
CPC分类号： C12N9/2417
摘要： The present invention provides a DNA fragment encoding alkaline liquefying α-amylase, recombinant DNA containing the DNA fragment, a transformed microorganism harboring the recombinant DNA, as well as a method for producing alkaline liquefying α-amylase using the transformant. The method of the present invention enables mass production of alkaline liquefying α-amylase useful as a detergent component.The present invention is directed to a liquefying alkaline alphα-amylase, and a DNA encoding for the same and functional fragments thereof.
摘要翻译：本发明提供了编码碱液化α-淀粉酶的DNA片段，含有DNA片段的重组DNA，携带重组DNA的转化微生物以及使用该转化体产生碱液化α-淀粉酶的方法。本发明的方法能够大量生产用作洗涤剂组分的碱性液化α-淀粉酶。本发明涉及液化碱性α-淀粉酶和编码其相同功能片段的DNA。

4. 发明授权

US06803222B2 Alkaline proteases 有权
标题翻译：碱性蛋白酶
公开(公告)号：US06803222B2
公开(公告)日：2004-10-12
申请号：US09985689
申请日：2001-11-05
申请人： Yuji Hatada , Akinori Ogawa , Yasushi Kageyama , Tsuyoshi Sato , Hiroyuki Araki , Nobuyuki Sumitomo , Mitsuyoshi Okuda , Katsuhisa Saeki
发明人： Yuji Hatada , Akinori Ogawa , Yasushi Kageyama , Tsuyoshi Sato , Hiroyuki Araki , Nobuyuki Sumitomo , Mitsuyoshi Okuda , Katsuhisa Saeki
IPC分类号： C12N948
CPC分类号： C12N9/54 , C11D3/38609 , C11D3/38618
摘要： The present invention relates to alkaline proteases having high specific activity and strong oxidant resistance. The present invention also relates to alkaline proteases having excellent detergency that is to be added to a detergent.
摘要翻译：本发明涉及具有高比活性和强氧化剂抗性的碱性蛋白酶。本发明还涉及具有优异的去污力的碱性蛋白酶，其被添加到洗涤剂中。

5. 发明授权

US09217139B2 Extracellularly secreted nuclease 有权
标题翻译：细胞外分泌的核酸酶
公开(公告)号：US09217139B2
公开(公告)日：2015-12-22
申请号：US13823990
申请日：2011-09-15
申请人： Kozue Mori , Yukari Ohta , Yuji Hatada , Nobuyuki Nakamura , Masayuki Miyazaki
发明人： Kozue Mori , Yukari Ohta , Yuji Hatada , Nobuyuki Nakamura , Masayuki Miyazaki
IPC分类号： C12N9/22 , C12R1/465 , C12Q1/44 , C12N1/08
CPC分类号： C12N9/22 , C12N1/08 , C12Q1/44 , C12R1/465
摘要： The purpose of the present invention is to provide a nuclease that secretes natural nonpathogenic microorganisms extracellularly, has higher specific activity than conventional nucleases, and is useful in nucleolytic degradation on an industrial scale. This purpose is achieved with an extracellularly secreted nuclease derived from Streptomyces bacteria, the nuclease having specific activity equal to or greater than the specific activity of Benzonase® when supplied to double-stranded DNA for 30 minutes at 37° C. in 20 mM Tris/HCl (pH 8.5) containing 1 mM MgCl2 and 1 mM CaCl2 after purification, using double-stranded DNA, single-stranded DNA, and RNA as substrates.
摘要翻译：本发明的目的是提供一种在细胞外分泌天然非致病微生物的核酸酶，比常规核酸酶具有更高的比活性，并且可用于工业规模的核酸分解降解。该目的通过来自链霉菌细菌的细胞外分泌的核酸酶实现，所述核酸酶在37℃下在20mM Tris / HCl中提供至双链DNA 30分钟时具有等于或大于Benzonase的比活性的比活性，纯化后含有1mM MgCl 2和1mM CaCl 2的HCl（pH8.5），使用双链DNA，单链DNA和RNA作为底物。

6. 发明授权

US08790907B2 Method for detachment and preparation of living microorganisms 有权
标题翻译：活细菌分离和制备方法
公开(公告)号：US08790907B2
公开(公告)日：2014-07-29
申请号：US13359797
申请日：2012-01-27
申请人： Sumihiro Koyama , Tadashi Maruyama , Chiaki Kato , Yuichi Nogi , Yuji Hatada , Yukari Ohta , Masaaki Konishi , Taishi Tsubouchi
发明人： Sumihiro Koyama , Tadashi Maruyama , Chiaki Kato , Yuichi Nogi , Yuji Hatada , Yukari Ohta , Masaaki Konishi , Taishi Tsubouchi
IPC分类号： C12N1/00 , C12N1/04 , C12N11/14
CPC分类号： C12N13/00 , C12N11/14
摘要： A method for immobilizing living microorganisms includes a step (1) of disposing a solution containing microorganisms as an electrolyte on the surface of a substrate at least one portion of which is an electrode, and applying a constant potential to the electrode to cause at least a portion of the microorganisms to attach to the surface of the substrate. The constant potential in step (1) is greater than −0.5 V but not greater than −0.2 V (vs Ag/AgCl) or greater than +0.2 V but not greater than +0.4 V (vs Ag/AgCl). The electrolyte in step (1) does not contain a source of nutrition for the microorganisms.
摘要翻译：固定活体微生物的方法包括将含有微生物作为电解质的溶液设置在基材的表面上的步骤（1），其中至少一部分是电极，并向电极施加恒定电位，使至少部分微生物附着到基底表面。步骤（1）中的恒定电位大于-0.5V但不大于-0.2V（相对于Ag / AgCl）或大于+ 0.2V但不大于+0.4V（相对于Ag / AgCl）。步骤（1）中的电解质不含微生物的营养来源。

7. 发明申请

US20110045570A1 NOVEL HIGH ALKALINE PROTEASE AND USE THEREOF 失效
标题翻译：新型高碱性蛋白酶及其用途
公开(公告)号：US20110045570A1
公开(公告)日：2011-02-24
申请号：US11719713
申请日：2005-11-16
申请人： Jie Lu , Zhijun Li , Si Hung Vo , Yuji Hatada , Ken Takai , Susumu Ito , Koki Horikoshi
发明人： Jie Lu , Zhijun Li , Si Hung Vo , Yuji Hatada , Ken Takai , Susumu Ito , Koki Horikoshi
IPC分类号： C12N9/52 , C12N9/50 , C07H21/04 , C07H21/00 , C12N15/63 , C12N1/21
CPC分类号： C12N9/52
摘要： The invention aims to provide a novel alkaline protease having peculiar properties such as high alkali activity, resistance to surfactants and calcium-dependent thermostability and exhibiting excellent performance in highly alkaline detergents, and a gene coding for the amino acid sequence thereof. There is provided an alkaline protease with such properties that an active pH range is from 5 to 13, an optimum pH is approximately 12.6, an optimum temperature is 70° C., no activity drop by heating is observed up to 65° C. at pH 10 and the optimum temperature and the thermostability are not affected by Ca2+ ions. Specifically, there is provided, for example, an alkaline protease having an amino acid sequence constituting a mature enzyme as represented by SEQ ID NO: 3 or an amino acid sequence resulting from deletion, substitution, situs inversus arrangement, addition or insertion of a part of amino acids thereof, or derived from Alkaliphillus transvaalensis. The protease cleaves 26 peptide bonds among 29 peptide bonds of acidic insulin B-chain.
摘要翻译：本发明的目的在于提供一种具有高碱性，耐表面活性和钙依赖性热稳定性等特性，在高碱性洗涤剂中表现出优异性能的新型碱性蛋白酶及其氨基酸序列的编码基因。提供了具有这样的性质的碱性蛋白酶，其活性pH范围为5至13，最佳pH为约12.6，最适温度为70℃，在65℃下不观察到加热下的活性降低 pH 10，最佳温度和热稳定性不受Ca2 +离子的影响。具体地，例如提供了具有SEQ ID NO：3所示的构成成熟酶的氨基酸序列的氨基酸序列的碱性蛋白酶或由缺失，取代，位置反转排列，添加或插入部分的氨基酸，或衍生自变形白菜（Alkaliphillus transvaalensis）。蛋白酶在酸性胰岛素B链的29个肽键中切割26个肽键。

8. 发明授权

US06486113B1 Mutant &agr;-amylases 失效
标题翻译：突变型α-淀粉酶
公开(公告)号：US06486113B1
公开(公告)日：2002-11-26
申请号：US09381687
申请日：1999-09-23
申请人： Yuji Hatada , Kaori Ikawa , Susumu Ito , Kazuaki Igarashi , Hiroshi Hagihara , Yasuhiro Hayashi , Hiroyuki Araki , Katsuya Ozaki
发明人： Yuji Hatada , Kaori Ikawa , Susumu Ito , Kazuaki Igarashi , Hiroshi Hagihara , Yasuhiro Hayashi , Hiroyuki Araki , Katsuya Ozaki
IPC分类号： C11D3386
CPC分类号： C12N9/2417 , C11D3/386
摘要： The invention relates to a mutant &agr;-amylase having an amino acid sequence obtained by making deletion or replacement by another arbitrary amino acid residue of at least a methionine residue at the 202-position or a position homologous thereto among amino acid residues set forth in SEQ ID NO:1, which constitute a liquefying alkaline &agr;-amylase, a gene thereof, and a detergent composition comprising the mutant &agr;-amylase. The mutant &agr;-amylase has the optimum pH in an alkaline range, an excellent &agr;-amylase activity, and high and lasting resistance to oxidizing agents, and is hence particularly useful as a component of detergent compositions containing a bleaching agent and an oxidizing agent.
摘要翻译：本发明涉及一种具有氨基酸序列的突变型α-淀粉酶，该氨基酸序列是通过在SEQ ID NO：1所示的氨基酸残基之间至少含有202位的甲硫氨酸残基或其同源位置的另外任意氨基酸残基进行缺失或替换而获得的 ID NO：1，其构成液化碱性α-淀粉酶，其基因和包含突变型α-淀粉酶的洗涤剂组合物。突变型α-淀粉酶在碱性范围内具有最佳pH，优异的α-淀粉酶活性和对氧化剂的高持久耐受性，因此特别可用作含有漂白剂和氧化剂的洗涤剂组合物的组分。

9. 发明授权

US06338959B1 Gene for enzyme having both alkaline pullulanase and alkaline &agr;-amylase activities 失效
标题翻译：具有碱性支链淀粉酶和碱性α-淀粉酶活性的酶的基因
公开(公告)号：US06338959B1
公开(公告)日：2002-01-15
申请号：US09514302
申请日：2000-02-28
申请人： Yuji Hatada , Kazuaki Igarashi , Katsuya Ozaki , Katsutoshi Ara , Shuji Kawai , Susumu Ito
发明人： Yuji Hatada , Kazuaki Igarashi , Katsuya Ozaki , Katsutoshi Ara , Shuji Kawai , Susumu Ito
IPC分类号： C12N928
CPC分类号： C12N9/2457 , C12N9/2417 , C12N9/2451 , C12Y302/01001 , C12Y302/01041
摘要： The present invention provides a DNA fragment encoding alkaline pullulanase exhibiting alkaline &agr;-amylase activity, alkaline &agr;-amylase possessing both an amino acid sequence described in SEQ ID NO:3 and a DNA fragment encoding the amylase, alkaline pullulanase possessing both an amino acid sequence described in SEQ ID NO:4 and a DNA fragment encoding the pullulanase, recombinant DNAs containing these DNA fragments, and transformed microorganisms harboring the recombinant DNAs. The technique of the present invention enables mass production of alkaline pullulanase exhibiting alkaline &agr;-amylase activity.
摘要翻译：本发明提供了编码具有碱性α-淀粉酶活性的碱性支链淀粉酶的DNA片段，具有SEQ ID NO：3所示的氨基酸序列的碱性α-淀粉酶和编码具有氨基酸序列的淀粉酶，碱基支链淀粉酶的DNA片段和编码支链淀粉酶的DNA片段，含有这些DNA片段的重组DNA和含有重组DNA的转化微生物。本发明的技术能够大量生产显示碱性α-淀粉酶活性的碱性支链淀粉酶。

10. 发明申请

US20110014677A1 NOVEL DNA FRAGMENT, RECOMBINANT VECTOR COMPRISING SAME, TRANSFORMANT TRANSFORMED THEREWITH, AND USE THEREOF 失效
标题翻译：新型DNA片段，包含其的重组载体，其变形变体及其用途
公开(公告)号：US20110014677A1
公开(公告)日：2011-01-20
申请号：US12452648
申请日：2008-07-09
申请人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
发明人： Yuji Hatada , Yukari Ohta , Yuko Hidaka , Nobuyuki Nakamura
IPC分类号： C12N9/02 , C12N9/10 , C12N9/14 , C12N9/88 , C12N9/90 , C12N1/21 , C12N15/63 , C07H21/04
CPC分类号： C12N15/63 , C07K2319/02 , C12N9/2437 , C12N9/2468 , C12N15/70 , C12N15/75 , C12P21/02 , C12Y302/01004 , C12Y302/01081
摘要： A DNA fragment containing a gene which encodes a specific gene regulatory region alone or the gene regulatory region together with a signal peptide; a recombinant vector containing the DNA fragment; a transformant containing the recombinant vector; and a method of producing a recombinant protein by using the transformant. According to the invention, it is possible to produce a protein in a large amount at a high efficiency regardless of the kind of the recombinant protein.
摘要翻译：含有单独编码特定基因调控区域的基因或与信号肽一起的基因调控区域的DNA片段; 含有该DNA片段的重组载体; 含有重组载体的转化体; 以及使用该转化体生产重组蛋白的方法。根据本发明，无论重组蛋白质的种类如何，都可以高效率地生产蛋白质。

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