公开(公告)号：US07179964B2

公开(公告)日：2007-02-20

申请号：US10194885

申请日：2002-07-12

申请人： Myeong-Je Cho ,  Joshua H. Wong ,  Peggy G. Lemaux ,  Bob B. Buchanan

发明人： Myeong-Je Cho ,  Joshua H. Wong ,  Peggy G. Lemaux ,  Bob B. Buchanan

IPC分类号： C12N15/82 ,  A01H5/00

CPC分类号： A21D13/064 ,  A23J1/12 ,  A23K10/30 ,  A23L33/105 ,  A23V2002/00 ,  C07K14/415 ,  C12N9/0036 ,  C12N15/8242 ,  C12N15/8245 ,  C12N15/8251 ,  C12N15/8261 ,  Y02A40/146 ,  A23V2300/21

摘要： The present invention is directed to a transgenic plant wherein at least a part of said plant includes a recombinant nucleic acid with a promoter active in the part operably linked to a nucleic acid encoding a thioredoxin polypeptide wherein the promoter is a seed or grain maturation-specific promoter and the thioredoxin polypeptide includes the amino acid sequence WCGPC. The present invention is further directed to transgenic plants that overexpress thioredoxin in seed wherein the overexpression of thioredoxin h effects a significant increase in the reduction of proteins (—SH as compared to S—S) of the albumin fraction of the seed.

摘要翻译： 本发明涉及一种转基因植物，其中所述植物的至少一部分包括具有启动子活性的启动子的重组核酸，其与编码硫氧还蛋白多肽的核酸可操作地连接，其中所述启动子是种子或谷物成熟特异性 启动子和硫氧还蛋白多肽包括氨基酸序列WCGPC。 本发明进一步涉及在种子中过表达硫氧还蛋白的转基因植物，其中硫氧还蛋白的过表达h与种子的白蛋白部分的蛋白质的降低（-SH）相比显着增加。

公开(公告)号：US6114504A

公开(公告)日：2000-09-05

申请号：US483930

申请日：1995-06-07

申请人： Bob B. Buchanan ,  Karoly Kobrehel ,  Boihon C. Yee ,  Joshua H. Wong ,  Rosa Lozano ,  Jin-an Jiao ,  Sungho Shin

发明人： Bob B. Buchanan ,  Karoly Kobrehel ,  Boihon C. Yee ,  Joshua H. Wong ,  Rosa Lozano ,  Jin-an Jiao ,  Sungho Shin

IPC分类号： A23L1/172 ,  A21D2/26 ,  A21D8/04 ,  A21D10/00 ,  A23C9/12 ,  A23J1/12 ,  A23J1/14 ,  A23J3/18 ,  A23L1/015 ,  A23L1/16 ,  A23L1/18 ,  A23L1/182 ,  A23L1/211 ,  A23L1/314 ,  A61K35/58 ,  A61K35/64 ,  A61K38/44 ,  A61P39/02 ,  C07K1/113 ,  C07K14/415 ,  C07K14/435 ,  C07K14/46 ,  C07K14/76 ,  C07K14/81 ,  C12N1/19 ,  C12N9/02 ,  C12N9/20 ,  C12N9/96 ,  C12N9/99 ,  C12N11/02 ,  C12P21/02 ,  C07K14/42 ,  A23J3/34 ,  A23J3/14

CPC分类号： C12P21/02 ,  A21D2/26 ,  A21D2/265 ,  A21D2/266 ,  A21D8/04 ,  A23C9/1213 ,  A23J1/12 ,  A23J1/14 ,  A23J3/18 ,  A23L11/34 ,  A23L13/48 ,  A23L5/27 ,  A23L7/109 ,  A23L7/196 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12Y108/01008 ,  C12Y108/01009 ,  A61K38/00

摘要： Methods of reducing cystine containing animal and plant proteins, and improving dough and baked goods' characteristics is provided which includes the steps of mixing dough ingredients with a thiol redox protein to form a dough and baking the dough to form a baked good. The method of the present invention preferably uses reduced thioredoxin with wheat flour which imparts a stronger dough and higher loaf volumes. Methods for reducing snake, bee and scorpion toxin proteins with a thiol redox (SH) agent and thereby inactivating the protein or detoxifying the protein in an individual are also provided. Protease inhibitors, including the Kunitz and Bowman-Birk trypsin inhibitors of soybean, were also reduced by the NADP/thioredoxin system (NADPH, thioredoxin, and NADP-thioredoxin reductase) from either E. coli or wheat germ. When reduced by thioredoxin, the Kunitz and Bowman-Birk soybean trypsin inhibitors lose their ability to inhibit trypsin. Moreover, the reduced form of the inhibitors showed increased susceptibility to heat and proteolysis by either subtilisin or a protease preparation from germinating wheat seeds. The 2S albumin of castor seed endosperm was reduced by thioredoxin from either wheat germ or E. coli. Thioredoxin was reduced by either NADPH and NADP-thioredoxin reductase or dithiothreitol. Analyses showed that thioredoxin actively reduced the intramolecular disulfides of the 2S large subunit, but was ineffective in reducing the intermolecular disulfides that connect the large to the small subunit. A novel cystine containing protein that inhibits pullulanase was isolated. The protein was reduced by thioredoxin and upon reduction its inhibitory activity was destroyed or greatly reduced.

公开(公告)号：US06838113B1

公开(公告)日：2005-01-04

申请号：US09296927

申请日：1999-04-22

申请人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Jin-an Jiao ,  Joshua H. Wong ,  Boihon C. Yee

发明人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Jin-an Jiao ,  Joshua H. Wong ,  Boihon C. Yee

IPC分类号： A21D2/26 ,  A21D2/36 ,  A21D8/04 ,  A21D13/06 ,  A23C9/12 ,  A23C9/152 ,  A23C9/20 ,  A23J1/12 ,  A23J1/14 ,  A23J3/08 ,  A23J3/16 ,  A23J3/18 ,  A23K1/16 ,  A23L1/015 ,  A23L1/16 ,  A23L1/182 ,  A23L1/211 ,  A23L1/305 ,  A23L1/314 ,  A61K38/44 ,  A61K39/35 ,  C07K1/113 ,  C07K14/415 ,  C07K14/435 ,  C07K14/46 ,  C07K14/76 ,  C07K14/81 ,  C12N9/02 ,  C12N9/20 ,  C12P21/02 ,  C12Q1/26 ,  C12Q1/34 ,  A23L1/29 ,  A23L3/358

CPC分类号： A21D13/064 ,  A21D2/26 ,  A21D2/264 ,  A21D2/265 ,  A21D2/266 ,  A21D2/36 ,  A21D8/04 ,  A23C9/1209 ,  A23C9/1213 ,  A23C9/152 ,  A23J1/12 ,  A23J1/14 ,  A23J3/08 ,  A23J3/18 ,  A23K20/147 ,  A23L5/27 ,  A23L7/109 ,  A23L7/196 ,  A23L11/34 ,  A23L13/48 ,  A23L33/17 ,  A61K38/44 ,  A61K39/35 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/43504 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12P21/02 ,  C12Q1/26 ,  C12Q1/34 ,  C12Y108/01008 ,  C12Y108/01009 ,  A61K2300/00

摘要： Thioredoxin, a small dithiol protein, is a specific reductant for major food proteins, allergenic proteins and particularly allergenic proteins present in widely used foods from animal and plant sources. All targeted proteins contain disulfide (S—S) bonds that are reduced to the sulfhydryl (SH) level by thioredoxin. The proteins are allergenically active and less digestible in the oxidized (S—S) state. When reduced (SH state), they lose their allergenicity and/or become more digestible. Thioredoxin achieved this reduction when activated (reduced) either by NADPH via NADP-thioredoxin reductase (physiological conditions) or by dithiothreitol, a chemical reductant. Skin tests and feeding experiments carried out with sensitized dogs showed that treatment of the food with reduced thioredoxin prior to ingestion eliminated or decreased the allergenicity of the food. Studies showed increased digestion of food and food proteins by pepsin and trypsin following reduction by thioredoxin.

公开(公告)号：US5952034A

公开(公告)日：1999-09-14

申请号：US953703

申请日：1997-10-17

申请人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Jin-an Jiao ,  Joshua H. Wong ,  Boihon C. Yee

发明人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Jin-an Jiao ,  Joshua H. Wong ,  Boihon C. Yee

IPC分类号： A21D2/26 ,  A21D2/36 ,  A21D8/04 ,  A21D13/06 ,  A23C9/12 ,  A23C9/152 ,  A23C9/20 ,  A23J1/12 ,  A23J1/14 ,  A23J3/08 ,  A23J3/16 ,  A23J3/18 ,  A23K1/16 ,  A23L1/015 ,  A23L1/16 ,  A23L1/182 ,  A23L1/211 ,  A23L1/305 ,  A23L1/314 ,  A61K38/44 ,  A61K39/35 ,  C07K1/113 ,  C07K14/415 ,  C07K14/435 ,  C07K14/46 ,  C07K14/76 ,  C07K14/81 ,  C12N9/02 ,  C12N9/20 ,  C12P21/02 ,  C12Q1/26 ,  C12Q1/34 ,  A23L1/03

CPC分类号： A21D13/064 ,  A21D2/26 ,  A21D2/264 ,  A21D2/265 ,  A21D2/266 ,  A21D2/36 ,  A21D8/04 ,  A23C9/1209 ,  A23C9/1213 ,  A23C9/152 ,  A23J1/12 ,  A23J1/14 ,  A23J3/08 ,  A23J3/18 ,  A23K20/147 ,  A23L11/34 ,  A23L13/48 ,  A23L33/17 ,  A23L5/27 ,  A23L7/109 ,  A23L7/196 ,  A61K38/44 ,  A61K39/35 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/43504 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12P21/02 ,  C12Q1/26 ,  C12Q1/34 ,  C12Y108/01008 ,  C12Y108/01009

摘要： Thioredoxin, a small dithiol protein, is a specific reductant for major food proteins, allergenic proteins and particularly allergenic proteins present in widely used foods from animal and plant sources. All targeted proteins contain disulfide (S--S) bonds that are reduced to the sulfhydryl (SH) level by thioredoxin. The proteins are allergenically active and less digestible in the oxidized (S--S) state. When reduced (SH state), they lose their allergenicity and/or become more digestible. Thioredoxin achieved this reduction when activated (reduced) either by NADPH via NADP-thioredoxin reductase (physiological conditions) or by dithiothreitol, a chemical reductant. Skin tests and feeding experiments carried out with sensitized dogs showed that treatment of the food with reduced thioredoxin prior to ingestion eliminated or decreased the allergenicity of the food. Studies showed increased digestion of food and food proteins by pepsin and trypsin following reduction by thioredoxin.

摘要翻译： 硫氧还蛋白，一种小二硫醇蛋白，是主要的食品蛋白质，过敏原蛋白质，特别是来自动物和植物来源广泛使用的食品中的过敏原蛋白质的特异性还原剂。 所有靶向蛋白含有通过硫氧还蛋白还原成巯基（SH）水平的二硫键（S-S）键。 蛋白质是过敏性活性的，在氧化（S-S）状态下消化不良。 当减少（SH状态）时，它们失去过敏原性和/或变得更易消化。 当NADPH通过NADP-硫氧还蛋白还原酶（生理条件）或二硫苏糖醇（化学还原剂）活化（还原）时，硫氧还蛋白实现了这种还原。 用敏化狗进行的皮肤试验和喂养实验表明，在摄取之前用硫氧还蛋白降低的食物处理消除或降低了食物的过敏原性。 研究显示硫氧还蛋白还原后胃蛋白酶和胰蛋白酶消化食物和食物蛋白质。

公开(公告)号：US06600021B1

公开(公告)日：2003-07-29

申请号：US09448111

申请日：1999-11-23

申请人： Bob B. Buchanan ,  Karoly Kobrehel ,  Boihon C. Yee ,  Joshua H. Wong ,  Rosa Lozano ,  Jin-an Jiao ,  Sungho Shin

发明人： Bob B. Buchanan ,  Karoly Kobrehel ,  Boihon C. Yee ,  Joshua H. Wong ,  Rosa Lozano ,  Jin-an Jiao ,  Sungho Shin

IPC分类号： C07K14415

CPC分类号： C12P21/02 ,  A21D2/26 ,  A21D2/265 ,  A21D2/266 ,  A21D8/04 ,  A23C9/1213 ,  A23J1/12 ,  A23J1/14 ,  A23J3/18 ,  A23L5/27 ,  A23L7/109 ,  A23L7/196 ,  A23L11/34 ,  A23L13/48 ,  A61K38/00 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12Y108/01008 ,  C12Y108/01009 ,  Y02A50/473 ,  A61K38/44 ,  A61K2300/00

摘要： Methods of reducing cystine containing animal and plant proteins, and improving dough and baked goods' characteristics is provided which includes the steps of mixing dough ingredients with a thiol redox protein to form a dough and baking the dough to form a baked good. The method of the present invention preferably uses reduced thioredoxin with wheat flour which imparts a stronger dough and higher loaf volumes. Methods for reducing snake, bee and scorpion toxin proteins with a thiol redox (SH) agent and thereby inactivating the protein or detoxifying the protein in an individual are also provided. Protease inhibitors, including the, Kunitz and Bowman-Birk trypsin inhibitors of soybean, were also reduced by the NADP/thioredoxin system (NADPH, thioredoxin, and NADP-thioredoxin reductase) from either E. coli or wheat germ. When reduced by thioredoxin, the Kunitz and Bowman-Birk soybean trypsin inhibitors lose their ability to inhibit trypsin. Moreover, the reduced form of the inhibitors showed increased susceptibility to heat and proteolysis by either subtilisin or a protease preparation from germinating wheat seeds. The 2S albumin of castor seed endosperm was reduced by thioredoxin from either wheat germ or E. coli. Thioredoxin was reduced by either NADPH and NADP-thioredoxin reductase or dithiothreitol. Analyses showed that thioredoxin actively reduced the intramolecular disulfides of the 2S large subunit, but was ineffective in reducing the intermolecular disulfides that connect the large to the small subunit. A novel cystine containing protein that inhibits pullulanase was isolated. The protein was reduced by thioredoxin and upon reduction its inhibitory activity was destroyed or greatly reduced.

公开(公告)号：US07074900B2

公开(公告)日：2006-07-11

申请号：US10422422

申请日：2003-04-23

申请人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Joshua H. Wong ,  Boihan C. Yee ,  Oscar L. Frick

发明人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Joshua H. Wong ,  Boihan C. Yee ,  Oscar L. Frick

IPC分类号： C07K14/00 ,  C07K14/415 ,  A61K39/35 ,  A61K39/36

CPC分类号： A21D2/26 ,  A21D2/264 ,  A21D2/265 ,  A21D2/266 ,  A21D2/36 ,  A21D8/04 ,  A21D13/064 ,  A23C9/1209 ,  A23C9/1213 ,  A23C9/152 ,  A23J1/12 ,  A23J1/14 ,  A23J3/08 ,  A23J3/18 ,  A23K20/147 ,  A23L5/27 ,  A23L7/109 ,  A23L11/34 ,  A23L13/48 ,  A23L33/17 ,  A61K39/35 ,  A61K2035/122 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/43504 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12P21/02 ,  C12Q1/26 ,  C12Q1/34 ,  C12Y108/01008 ,  C12Y108/01009 ,  Y10S436/904 ,  Y10T436/182

摘要： Thioredoxin, a small dithiol protein, is a specific reductant for allergenic proteins and particularly allergenic proteins present in pollen and animal and plant sources. All targeted proteins contain disulfide (S—S) bonds that are reduced to the sulfhydryl (SH) level by thioredoxin. The proteins are allergenically active and less digestible in the oxidized (S—S) state. When reduced (SH state), they lose their allergenicity and/or become more digestible. Thioredoxin achieved this reduction when activated (reduced) either by NADPH via NADP-thioredoxin reductase (physiological conditions) or by lipoic acid chemical reductant. Skin tests carried out with sensitized dogs showed that treatment of the pollens with reduced thioredoxin prior to injection eliminated or decreased the allergenicity of the pollen. Studies showed increased digestion of the pollen proteins by pepsin following reduction by thioredoxin. Pollen proteins that have been reduced by thioredoxin are effective and safe immunotherapeutic agents for decreasing or eliminating an animal's allergic reaction that would otherwise occur upon exposure to the non-reduced pollen protein.

公开(公告)号：US06555116B1

公开(公告)日：2003-04-29

申请号：US09238379

申请日：1999-01-27

申请人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Joshua H. Wong ,  Boihon C. Yee ,  Oscar L. Frick

发明人： Bob B. Buchanan ,  Gregorio del Val ,  Rosa M. Lozano ,  Joshua H. Wong ,  Boihon C. Yee ,  Oscar L. Frick

IPC分类号： A61K3936

CPC分类号： A21D2/26 ,  A21D2/264 ,  A21D2/265 ,  A21D2/266 ,  A21D2/36 ,  A21D8/04 ,  A21D13/064 ,  A23C9/1209 ,  A23C9/1213 ,  A23C9/152 ,  A23J1/12 ,  A23J1/14 ,  A23J3/08 ,  A23J3/18 ,  A23K20/147 ,  A23L5/27 ,  A23L7/109 ,  A23L11/34 ,  A23L13/48 ,  A23L33/17 ,  A61K39/35 ,  A61K2035/122 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/43504 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12P21/02 ,  C12Q1/26 ,  C12Q1/34 ,  C12Y108/01008 ,  C12Y108/01009 ,  Y10S436/904 ,  Y10T436/182

摘要： Thioredoxin, a small dithiol protein, is a specific reductant for allergenic proteins and particularly allergenic proteins present in pollen and animal and plant sources. All targeted proteins contain disulfide (S—S) bonds that are reduced to the sulfhydryl (SH) level by thioredoxin. The proteins are allergenically active and less digestible in the oxidized (S—S) state. When reduced (SH state), they lose their allergenicity and/or become more digestible. Thioredoxin achieved this reduction when activated (reduced) either by NADPH via NADP-thioredoxin reductase (physiological conditions) or by lipoic acid chemical reductant. Skin tests carried out with sensitized dogs showed that treatment of the pollens with reduced thioredoxin prior to injection eliminated or decreased the allergenicity of the pollen. Studies showed increased digestion of the pollen proteins by pepsin following reduction by thioredoxin. Pollen proteins that have been reduced by thioredoxin are effective and safe immunotherapeutic agents for decreasing or eliminating an animal's allergic reaction that would otherwise occur upon exposure to the non-reduced pollen protein.

摘要翻译： 硫氧还蛋白，一种小二硫醇蛋白，是一种特异性的还原剂，适用于过敏原蛋白，特别是花粉和动植物来源的变应原蛋白。 所有靶向蛋白含有通过硫氧还蛋白还原成巯基（SH）水平的二硫键（S-S）键。 蛋白质是过敏性活性的，在氧化（S-S）状态下消化不良。 当减少（SH状态）时，它们失去过敏原性和/或变得更易消化。 当NADPH通过NADP-硫氧还蛋白还原酶（生理条件）或硫辛酸化学还原剂被活化（还原）时，硫氧还蛋白实现了这一还原。 用致敏狗进行的皮肤试验表明，在注射前用降低的硫氧还蛋白处理花粉消除或降低了花粉的致敏性。 研究显示硫氧还蛋白还原后，通过胃蛋白酶消化花粉蛋白。 通过硫氧还蛋白还原的花粉蛋白是有效和安全的免疫治疗剂，用于减少或消除否则在暴露于未还原花粉蛋白时会发生的动物过敏反应。

公开(公告)号：US6113951A

公开(公告)日：2000-09-05

申请号：US211673

申请日：1994-11-21

申请人： Bob B. Buchanan ,  Karoly Kobrehel ,  Boihon C. Yee ,  Joshua H. Wong ,  Rosa Lozano ,  Jin-an Jiao ,  Sungho Shin

发明人： Bob B. Buchanan ,  Karoly Kobrehel ,  Boihon C. Yee ,  Joshua H. Wong ,  Rosa Lozano ,  Jin-an Jiao ,  Sungho Shin

IPC分类号： A23L1/172 ,  A21D2/26 ,  A21D8/04 ,  A21D10/00 ,  A23C9/12 ,  A23J1/12 ,  A23J1/14 ,  A23J3/18 ,  A23L1/015 ,  A23L1/16 ,  A23L1/18 ,  A23L1/182 ,  A23L1/211 ,  A23L1/314 ,  A61K35/58 ,  A61K35/64 ,  A61K38/44 ,  A61P39/02 ,  C07K1/113 ,  C07K14/415 ,  C07K14/435 ,  C07K14/46 ,  C07K14/76 ,  C07K14/81 ,  C12N1/19 ,  C12N9/02 ,  C12N9/20 ,  C12N9/96 ,  C12N9/99 ,  C12N11/02 ,  C12P21/02 ,  A21D13/06 ,  A23J3/14 ,  C12P21/06

CPC分类号： C12P21/02 ,  A21D2/26 ,  A21D2/265 ,  A21D2/266 ,  A21D8/04 ,  A23C9/1213 ,  A23J1/12 ,  A23J1/14 ,  A23J3/18 ,  A23L11/34 ,  A23L13/48 ,  A23L5/27 ,  A23L7/109 ,  A23L7/196 ,  C07K1/1133 ,  C07K14/415 ,  C07K14/46 ,  C07K14/76 ,  C07K14/811 ,  C12N9/0036 ,  C12N9/20 ,  C12Y108/01008 ,  C12Y108/01009 ,  A61K38/00

摘要： Methods of reducing cystine containing animal and plant proteins, and improving dough and baked goods' characteristics is provided which includes the steps of mixing dough ingredients with a thiol redox protein to form a dough and baking the dough to form a baked good. The method of the present invention preferably uses reduced thioredoxin with wheat flour which imparts a stronger dough and higher loaf volumes. Methods for reducing snake, bee and scorpion toxin proteins with a thiol redox (SH) agent and thereby inactivating the protein or detoxifying the protein in an individual are also provided. Protease inhibitors, including the Kunitz and Bowman-Birk trypsin inhibitors of soybean, were also reduced by the NADP/thioredoxin system (NADPH, thioredoxin, and NADP-thioredoxin reductase). When reduced by thioredoxin, the Kunitz and Bowman-Birk soybean trypsin inhibitors lose their ability to inhibit trypsin. Moreover, the reduced form of the inhibitors showed increased susceptibility to heat and proteolysis by either subtilisin or a protease preparation from germinating wheat seeds. The 2S albumin of castor seed endosperm was reduced by thioredoxin. Thioredoxin was reduced by either NADPH and NADP-thioredoxin reductase or dithiothreitol. Analyses showed that thioredoxin actively reduced the intramolecular disulfides of the 2S large subunit, but was ineffective in reducing the intermolecular disulfides that connect the large to the small subunit. A novel cystine containing protein that inhibits pullulanase was isolated; thioredoxin reduction of this protein destroyed or greatly reduced its inhibitory activity.