公开(公告)号：US06521444B1

公开(公告)日：2003-02-18

申请号：US09711508

申请日：2000-11-14

Applicant: Koichi Numata ,  Yasushi Oda ,  Masami Miyata ,  Yukio Okamura ,  Toshiaki Kimura ,  Masatoshi Uchida ,  Osamu Asami

Inventor： Koichi Numata ,  Yasushi Oda ,  Masami Miyata ,  Yukio Okamura ,  Toshiaki Kimura ,  Masatoshi Uchida ,  Osamu Asami

IPC: B09B300

CPC classification number: C12R1/01 ,  A62D3/02 ,  A62D2101/22 ,  B09C1/10 ,  C02F3/34 ,  C12P1/04

Abstract: A novel microorganism which has the following characteristics: morphology (coccoid, rod shaped); gram staining (+), spore forming (−), motility (−), relationship to oxygen (aerobic), oxidase test (−), catalase test (+), resistance to acid (−), rod-coccus cycle (+), and GC content of DNA (mole%) (73 (by HPLC)), and which can decompose chloroethylene. The microorganism can decompose in 24 hours 30 ppm of trichloroethylene, and decompose of 100 ppm of trichloroethylene by 50%.

Abstract translation: 具有以下特征的新型微生物：形态（coccoid，rod shaped）; 革兰氏染色（+），孢子形成（ - ），运动性（ - ），与氧的关系（需氧），氧化酶试验（ - ），过氧化氢酶试验（+），耐酸性（ - ），棒球菌循环 ，和GC含量（摩尔％）（73（通过HPLC）），并可分解氯乙烯。微生物可在24小时内分解30ppm的三氯乙烯，并将100ppm的三氯乙烯分解50％。

公开(公告)号：US06171844B2

公开(公告)日：2001-01-09

申请号：US09252806

申请日：1999-02-19

Applicant: Koichi Numata ,  Yasushi Oda ,  Masami Miyata ,  Yukio Okamura ,  Toshiaki Kimura ,  Masatoshi Uchida ,  Osamu Asami

Inventor： Koichi Numata ,  Yasushi Oda ,  Masami Miyata ,  Yukio Okamura ,  Toshiaki Kimura ,  Masatoshi Uchida ,  Osamu Asami

IPC: C12N120

CPC classification number: C12R1/01 ,  A62D3/02 ,  A62D2101/22 ,  B09C1/10 ,  C02F3/34 ,  C12P1/04

Abstract: A novel microorganism which has the following characteristics: morphology (coccoid, rod shaped), gram staining (+), spore forming (−), motility (−), relationship to exygen (aerobic), oxidase test (−), catalase test (+), resistance to acid (−), rod-coccus cycle (+), and GC content of DNA (mole %) (73 (by HPLC)), and which can decompose chloroethylene. The microorganism can decompose in 24 hours 30 ppm of trichloroethylene, and decompose of 100 ppm of trichloroethylene by 50%.

Abstract translation: 一种具有以下特征的新型微生物：形态（coccoid，rod shaped），革兰氏染色（+），孢子形成（ - ），运动（ - ），与exygen（好氧），氧化酶试验（ - ），过氧化氢酶试验 +），耐酸性（ - ），杆 - 球菌循环（+）和GC的GC含量（摩尔％）（73（通过HPLC）），并且其可以分解氯乙烯。 微生物可在24小时内分解30ppm的三氯乙烯，并将100ppm的三氯乙烯分解50％。

公开(公告)号：US4844897A

公开(公告)日：1989-07-04

申请号：US906240

申请日：1986-09-12

Applicant: Hiroshi Maeda ,  Yasuhiro Matsumura ,  Osamu Asami ,  Hideyuki Tanaka ,  Ikuharu Sasaki

Inventor： Hiroshi Maeda ,  Yasuhiro Matsumura ,  Osamu Asami ,  Hideyuki Tanaka ,  Ikuharu Sasaki

IPC: A61K38/00 ,  C12N9/52 ,  C12N9/54 ,  C12N9/96

CPC classification number: C12N9/54 ,  C12N9/52 ,  C12N9/96 ,  A61K38/00

Abstract: Method of treating a tumor in a mammal comprises administering to said mammal an effective anti-tumor amount of proteases originating from microorganisms.Method of treating a tumor in a mammal comprises administering to said mammal an effective anti-tumor amount of a protease originating from a microorganism which protease is chemically modified by one of the following procedures:(a) coupling with a saccharide,(b) introduction of a hydrophobic polymeric group,(c) alteration of electric charge of the protein surface,(d) conjugation with a low molecular weight anti-tumor agent of molecular weight less than 2,000,(e) formation of dimer or oligomer by cross-linking of protease molecules,(f) conjugation with a synthetic polycation,(g) conjugation with a synthetic polyanion, and(h) combination of the above-mentioned procedures.Microorganism protease is chemically modified by one of the following procedures:(a) coupling with a saccharide,(b) introduction of a hydrophobic polymeric group,(c) alteration of electric charge of the protein surface,(d) conjugation with a low molecular weight anti-tumor agent of molecular weight less than 2,000,(e) formation of dimer or oligomer by cross-linking of protease molecules,(f) conjugation with a synthetic polycation,(g) conjugation with a synthetic polyanion, and(h) combination of the above-mentioned procedures.

Abstract translation: 治疗哺乳动物肿瘤的方法包括向所述哺乳动物施用有效的抗肿瘤量的源于微生物的蛋白酶。 治疗哺乳动物肿瘤的方法包括向所述哺乳动物施用有效的抗肿瘤量的来自微生物的蛋白酶，所述蛋白酶通过以下方法之一进行化学修饰：（a）与糖的偶联，（b）引入 的疏水性聚合物组，（c）蛋白质表面电荷的改变，（d）与分子量小于2,000的低分子量抗肿瘤剂缀合，（e）通过交联形成二聚体或低聚物 的蛋白酶分子，（f）与合成聚阳离子的缀合，（g）与合成聚阴离子的缀合，和（h）上述步骤的组合。 微生物蛋白酶通过以下方法之一进行化学修饰：（a）与糖的偶联，（b）引入疏水聚合基团，（c）改变蛋白质表面的电荷，（d）与低分子 （e）通过蛋白酶分子的交联形成二聚体或寡聚体，（f）与合成聚阳离子的缀合，（g）与合成聚阴离子的缀合，和（h）分子量小于2000的重量抗肿瘤剂， 组合上述程序。

公开(公告)号：US5700659A

公开(公告)日：1997-12-23

申请号：US464365

申请日：1995-06-05

Applicant: Yukio Yamada ,  Osamu Asami ,  Hidehiko Sugiyama ,  Chie Idekoba ,  Fumihiko Hoshino ,  Masana Hirai ,  Tsutomu Kajino ,  Takao Imaeda ,  Kiyoko Sarai

Inventor： Yukio Yamada ,  Osamu Asami ,  Hidehiko Sugiyama ,  Chie Idekoba ,  Fumihiko Hoshino ,  Masana Hirai ,  Tsutomu Kajino ,  Takao Imaeda ,  Kiyoko Sarai

IPC: C12N9/90 ,  C12N15/52 ,  C12N15/31

CPC classification number: C12N9/90

Abstract: A highly thermostable polypeptide possessing protein disulfide isomerase (PDI) activity, a gene coding for the polypeptide and a process for producing the polyeptide are provided. The polypeptide possessing PDI activity is characterized by A) having a capability of catalyzing a disulfide exchange in proteins, B) recognizing mainly ribonuclease A as a substrate, C) having a suitable active temperature of 20.degree. to 70.degree. C., D) being stable at a pH value of 6 to 9, and E) having a molecular weight of about 60,000 to 62,000. Since it has a higher thermostability and exhibits a stable activity in a wider dithiothreitol concentration range as compared with the conventional PDI, it is possible to provide a novel enzyme active protein which can be advantageously used for a refolding reaction of certain proteins. Further, a process which enables the polylpeptide possessing PDI activity to be efficiently produced using Humicola insolens or a transformant transformed with an expression vector containing the above-described gene is also provided.

Abstract translation: 提供了具有蛋白质二硫键异构酶（PDI）活性的高度热稳定的多肽，编码多肽的基因和产生多肽的方法。 具有PDI活性的多肽的特征在于：A）具有催化蛋白质中二硫键交换的能力，B）主要以核糖核酸酶A为底物，C）具有20〜70℃的合适的活性温度，D）为 在pH值为6〜9的条件下稳定，E）分子量约为60,000〜62,000。 由于与常规PDI相比，具有较高的热稳定性并且在更宽的二硫苏糖醇浓度范围内表现出稳定的活性，因此可以提供可有利地用于某些蛋白质的重折叠反应的新型酶活性蛋白质。 此外，还提供了使用Humicola insolens或用含有上述基因的表达载体转化的转化体能够有效地产生具有PDI活性的多肽的方法。

公开(公告)号：US5998198A

公开(公告)日：1999-12-07

申请号：US981352

申请日：1997-12-19

Applicant: Mika Nakayama ,  Chie Miyazaki ,  Osamu Asami ,  Yukio Yamada ,  Koichi Numata ,  Yasushi Oda

Inventor： Mika Nakayama ,  Chie Miyazaki ,  Osamu Asami ,  Yukio Yamada ,  Koichi Numata ,  Yasushi Oda

IPC: A62D3/02 ,  B09C1/10 ,  C02F3/34 ,  C12P1/04 ,  B09B3/00

CPC classification number: C12R1/38 ,  B09C1/10 ,  C02F3/34 ,  C12P1/04 ,  C12R1/01 ,  Y10S435/874

Abstract: Microorganisms belonging to the genus Burkholderia and having the ability to decompose halogenated hydrocarbon, which are able to decompose 50% or more of 100 ppm of trichloroethylene in 2 days, or decompose 100% of 30 ppm of trichloroethylene in 18 hours, as well as providing a process for decomposing halogenated hydrocarbons in water or soil using those microorganisms.

Abstract translation: PCT No.PCT / JP97 / 01359 Sec。 371 1997年12月19日第 102（e）1997年12月19日PCT PCT 1997年4月18日PCT公布。 WO97 / 40136 PCT公开号 日期：1997年10月30日属于伯克霍尔德氏菌属的微生物，具有分解卤代烃的能力，其能够在2天内分解50ppm或更多的100ppm三氯乙烯，或在18小时内分解100ppm的三氯乙烯100％ ，以及提供使用这些微生物分解水或土壤中的卤代烃的方法。

公开(公告)号：US06410251B2

公开(公告)日：2002-06-25

申请号：US09440585

申请日：1999-11-15

Applicant: Fumihiko Hoshino ,  Osamu Asami ,  Hideo Nakane ,  Yukio Yamada

Inventor： Fumihiko Hoshino ,  Osamu Asami ,  Hideo Nakane ,  Yukio Yamada

IPC: G01N33543

CPC classification number: G01N33/581 ,  Y10S435/972 ,  Y10S435/975 ,  Y10S436/806 ,  Y10S436/819

Abstract: A method for detecting or assaying one constituting member in a specific binding pair, for example, the antigen in an antigen/antibody pair, by utilizing specific binding such as binding between an antigen and an antibody, together with redox reaction for detecting a label, wherein an oxygen micro-electrode with a sensing surface area of 1 mm2 or less is used; and an apparatus to which the method is applicable. According to the method and by using the apparatus, redox reaction for assaying the label can be completed in such a short time as several minutes. Therefore, an inexpensive disposable apparatus for household use can be realized.

Abstract translation: 通过利用特异性结合如抗原和抗体之间的结合以及用于检测标记的氧化还原反应来检测或测定特异性结合对中的一个构成成员，例如抗原/抗体对中的抗原的方法， 其中使用感测表面积为1mm 2以下的氧微电极; 以及适用该方法的装置。 根据该方法，通过使用该装置，用于测定标签的氧化还原反应可以在几分钟的短时间内完成。 因此，可以实现廉价的家庭用一次性装置。

公开(公告)号：US5728531A

公开(公告)日：1998-03-17

申请号：US624116

申请日：1996-03-29

Applicant: Yukio Yamada ,  Osamu Asami ,  Hiroko Uchiyama ,  Hidehiko Sugiyama ,  Satoshi Fujita ,  Takekazu Yamamoto ,  Naoto Kagiyama ,  Masayoshi Momiyama ,  Yasumitsu Kondoh

Inventor： Yukio Yamada ,  Osamu Asami ,  Hiroko Uchiyama ,  Hidehiko Sugiyama ,  Satoshi Fujita ,  Takekazu Yamamoto ,  Naoto Kagiyama ,  Masayoshi Momiyama ,  Yasumitsu Kondoh

IPC: G01N33/58 ,  C07H19/10 ,  C12N15/09 ,  C12Q1/68 ,  G01N33/53 ,  G01N37/00 ,  C07H21/04 ,  C12N15/00

CPC classification number: C12Q1/6804 ,  C07H19/10 ,  C12Q1/6834

Abstract: A method of detecting nucleic acid utilizes hybridization between the nucleic acid to be detected which is present on an immobilizing support and a DNA probe which is complementary to at least a portion of the desired nucleotide sequence of the nucleic acid. The present DNA probe includes a hapten attached via a linker which a hapten is gibberellin or a gibberellin derivative. After hybridization by reaction between the nucleic acid to be detected and the DNA probe, the unreacted DNA probe is removed, and then labelled anti-hapten antibody is allowed to bind with the DNA probe of the hybrid and the labelling is used to detect the nucleic acid. Accordingly, stable detecting sensitivity may be achieved regardless of the label mixing ratio or labelling ratio, and thus regardless of the nucleotide sequence composition of the nucleic acid being tested.

Abstract translation: 检测核酸的方法利用存在于固定化载体上的待检测核酸与与核酸的所需核苷酸序列的至少一部分互补的DNA探针之间的杂交。 本DNA探针包括通过接头连接的半抗原，半抗原是赤霉素或赤霉素衍生物。 在待检测核酸与DNA探针之间的反应杂交后，除去未反应的DNA探针，然后标记抗半抗原抗体与杂交体的DNA探针结合，并用标记物检测核酸 酸。 因此，无论标记混合比或标记比例如何，均可实现稳定的检测灵敏度，因此与所测核酸的核苷酸序列组成无关。

公开(公告)号：US5496719A

公开(公告)日：1996-03-05

申请号：US68395

申请日：1993-05-27

Applicant: Yukio Yamada ,  Osamu Asami ,  Hidehiko Sugiyama ,  Chie Idekoba ,  Fumihiko Hoshino ,  Masana Hirai ,  Tsutomu Kajino ,  Takao Imaeda ,  Kiyoko Sarai

Inventor： Yukio Yamada ,  Osamu Asami ,  Hidehiko Sugiyama ,  Chie Idekoba ,  Fumihiko Hoshino ,  Masana Hirai ,  Tsutomu Kajino ,  Takao Imaeda ,  Kiyoko Sarai

IPC: C12N9/90 ,  A61K38/00 ,  C07K1/00 ,  C12N9/00

CPC classification number: C12N9/90

Abstract: A highly thermostable polypeptide possessing protein disulfide isomerase (PDI) activity, a gene coding for the polypeptide and a process for producing the polypeptide are provided. The polypeptide possessing PDI activity is characterized by A) having a capability of catalyzing a disulfide exchange in proteins, B) recognizing mainly ribonuclease A as a substrate, C) having a suitable active temperature of 20.degree. to 70.degree. C., D) being stable at a pH value of 6 to 9, and E) having a molecular weight of about 60,000 to 62,000. Since it has a higher thermostability and exhibits a stable activity in a wider dithiothreitol concentration range as compared with the conventional PDI, it is possible to provide a novel enzyme active protein which can be advantageously used for a refolding reaction of certain proteins. Further, a process which enables the polypeptide possessing PDI activity to be efficiently produced using Humicola insolens or a transformant transformed with an expression vector containing the above-described gene is also provided.

Abstract translation: 提供了具有蛋白质二硫键异构酶（PDI）活性的高度热稳定的多肽，编码多肽的基因和产生多肽的方法。 具有PDI活性的多肽的特征在于：A）具有催化蛋白质中二硫键交换的能力，B）主要以核糖核酸酶A为底物，C）具有20〜70℃的合适的活性温度，D）为 在pH值为6〜9的条件下稳定，E）分子量约为60,000〜62,000。 由于与常规PDI相比，具有较高的热稳定性并且在更宽的二硫苏糖醇浓度范围内表现出稳定的活性，因此可以提供可有利地用于某些蛋白质的重折叠反应的新型酶活性蛋白质。 此外，还提供了能够使用Humicola insolens或使用含有上述基因的表达载体转化的转化体有效地产生具有PDI活性的多肽的方法。