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    Nucleic acid sequences and expression systems for heparinase II and heparinase III derived from Flavobacterium heparinum
    1.
    发明授权
    Nucleic acid sequences and expression systems for heparinase II and heparinase III derived from Flavobacterium heparinum 失效
    肝素肝素肝素II和肝素酶III的核酸序列和表达系统

    公开(公告)号：US5681733A

    公开(公告)日：1997-10-28

    申请号：US258639

    申请日：1994-06-10

    申请人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

    发明人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

    IPC分类号： C12N15/09 ,  C07K16/40 ,  C12N1/21 ,  C12N9/88 ,  C12N15/60 ,  C12N15/74 ,  C12P21/08 ,  C12R1/19

    CPC分类号： C12Y402/02007 ,  C12N15/74 ,  C12N9/88

    摘要： The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading enzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite chromatography is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I, II and III are described.

    摘要翻译： 本发明描述了编码肝素黄肝素和硫酸乙酰肝素降解酶，肝素酶II和肝素酶III（EC 4.2.2.8）的基因的分离和序列。 它进一步描述了使用衍生自肝素肝素的糖胺聚糖裂解酶基因的启动子的修饰的核糖体结合区域来表达和表达肝素酶I，II和III的方法。 此外，概述了掺入细胞破碎，阳离子交换层析，亲和层析和羟基磷灰石层析的多步蛋白质纯化方法。 描述了针对肝炎黄杆菌蛋白质共同的翻译后修饰部分的抗体以及获得对这些部分和肝素酶I，II和III的氨基酸序列特异性的抗体的方法。

    Nucleic acid sequences and expression systems for heparinase II and heparinase III derived from Flavobacterium heparinum
    2.
    发明授权
    Nucleic acid sequences and expression systems for heparinase II and heparinase III derived from Flavobacterium heparinum 失效
    肝素肝素肝素II和肝素酶III的核酸序列和表达系统

    公开(公告)号：US5919693A

    公开(公告)日：1999-07-06

    申请号：US900951

    申请日：1997-07-25

    申请人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

    发明人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

    IPC分类号： C12N15/09 ,  C07K16/40 ,  C12N1/21 ,  C12N9/88 ,  C12N15/60 ,  C12N15/74 ,  C12P21/08 ,  C12R1/19 ,  C12N15/63

    CPC分类号： C12Y402/02007 ,  C12N15/74 ,  C12N9/88

    摘要： The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading enzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite chromatography is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I, II and III are described.

    摘要翻译： 本发明描述了编码肝素黄肝素和硫酸乙酰肝素降解酶，肝素酶II和肝素酶III（EC 4.2.2.8）的基因的分离和序列。 它进一步描述了使用衍生自肝素肝素的糖胺聚糖裂解酶基因的启动子的修饰的核糖体结合区域来表达和表达肝素酶I，II和III的方法。 此外，概述了掺入细胞破碎，阳离子交换层析，亲和层析和羟基磷灰石层析的多步蛋白质纯化方法。 描述了针对肝炎黄杆菌蛋白质共同的翻译后修饰部分的抗体以及获得对这些部分和肝素酶I，II和III的氨基酸序列特异性的抗体的方法。

    Chondroitin lyase enzymes
    3.
    发明授权
    Chondroitin lyase enzymes 失效
    软骨素裂解酶

    公开(公告)号：US6093563A

    公开(公告)日：2000-07-25

    申请号：US272247

    申请日：1994-07-08

    申请人： D. Clark Bennett ,  Maryse Laliberte ,  Kangfu Gu ,  Joseph Zimmermann ,  Anna Lydia Tkalec ,  Dominique Fink ,  Robert Linhardt

    发明人： D. Clark Bennett ,  Maryse Laliberte ,  Kangfu Gu ,  Joseph Zimmermann ,  Anna Lydia Tkalec ,  Dominique Fink ,  Robert Linhardt

    IPC分类号： C12N15/09 ,  A61K38/00 ,  C12N9/88 ,  C12N15/60 ,  C12R1/20 ,  A61K38/51

    CPC分类号： C12N9/88

    摘要： The present invention describes a method for the production of two highly purified enzymes capable of degrading chondroitin sulfate polysaccharides. A multi-step purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography, hydroxylapatite chromatography, high resolution ion exchange chromatography and size exclusion is outlined. A 77,000.+-.5,000 Dalton protein capable of degrading chondroitin sulfates A and C and a 55,000.+-.2,300 Dalton protein capable of degrading dermatan sulfate were isolated. The genes encoding these enzymes, chondroitinase AC and chondroitinase B, respectively, have been cloned and methods for their use are described.

    摘要翻译： 本发明描述了生产能够降解硫酸软骨素多糖的两种高度纯化的酶的方法。 概述了包括细胞破碎，阳离子交换层析，亲和层析，羟基磷灰石层析，高分辨率离子交换层析和尺寸排除的多步纯化方法。 分离能够降解硫酸软骨素A和C的77,000 +/- 5,000道尔顿蛋白质和能够降解硫酸皮肤素的55,000 +/- 2300道尔顿蛋白质。 已经克隆了编码这些酶，软骨素酶AC和软骨素酶B的基因，并描述了其使用方法。

    Attenuation of wound healing processes
    5.
    发明授权
    Attenuation of wound healing processes 失效
    伤口愈合过程衰减

    公开(公告)号：US5997863A

    公开(公告)日：1999-12-07

    申请号：US273109

    申请日：1994-07-08

    申请人： Joseph Zimmermann ,  Israel Vlodavsky ,  Clark Bennett ,  Pamela Danagher ,  Richard Broughton

    发明人： Joseph Zimmermann ,  Israel Vlodavsky ,  Clark Bennett ,  Pamela Danagher ,  Richard Broughton

    IPC分类号： A61K9/00 ,  A61K9/70 ,  A61K38/46 ,  A61K38/51 ,  A61L15/44 ,  A61P9/08 ,  A61P17/02 ,  A61P25/30 ,  A61P43/00 ,  A61K38/47 ,  C12N9/26 ,  C12N9/88

    CPC分类号： A61K38/51 ,  Y10S530/825

    摘要： Glycosaminoglycans, including heparinases 1, 2 and 3 as well as chondroitinases AC and B from the Gram negative bacteria Flavobacterium heparinum, can be used either separately or in combination to manipulate cell proliferation. In one embodiment, heparinases are administered to degrade heparan sulfate components of the extracellular matrix, thereby allowing the heparin binding growth factors which are stored in the extracellular matrix to migrate to adjacent cells. The mobility of chemoattractant agents, growth factors and cells also can be increased by treating tissues with glycosaminoglycan degrading enzymes, both chondroitinases and heparinases. The enzymatic removal of chondroitin sulfates from cell surfaces effectively increases the availability of growth factor receptors on the cell's surface. Selectively removing heparan sulfate from cell surfaces while leaving the extracellular matrix intact, conversely, inhibits cell proliferation by down regulating the cell's response to growth factors. This is achieved by targeting heparin or heparan sulfate degrading activities to the cell surface. Targeting the heparin degrading activity can be achieved by genetically engineering a ligand binding functionality into the heparinase proteins, or by physically controlling the localized enzyme concentration through the method of administration.

    摘要翻译： 糖胺聚糖，包括肝素酶1,2和3以及来自革兰氏阴性细菌肝炎黄杆菌的软骨素酶AC和B可以单独使用或组合使用以操纵细胞增殖。 在一个实施方案中，施用肝素酶以降解细胞外基质的硫酸乙酰肝素成分，从而使存储在细胞外基质中的肝素结合生长因子迁移至相邻细胞。 通过用糖胺聚糖降解酶（软骨素酶和肝素酶）处理组织，也可以增加化学引诱剂，生长因子和细胞的迁移率。 从细胞表面去除硫酸软骨素有效地增加细胞表面生长因子受体的可利用性。 从细胞表面选择性地去除硫酸乙酰肝素，同时使细胞外基质完好无损，相反地，通过下调细胞对生长因子的反应来抑制细胞增殖。 这是通过将肝素或硫酸乙酰肝素降解活性靶向细胞表面来实现的。 靶向肝素降解活性可以通过将配体结合功能遗传工程化成肝素酶蛋白质，或通过通过施用方法物理控制局部酶浓度来实现。

    Use of heparinases to decrease inflammatory response
    7.
    发明申请
    Use of heparinases to decrease inflammatory response 失效
    使用肝素降低炎症反应

    公开(公告)号：US20060140928A1

    公开(公告)日：2006-06-29

    申请号：US11357967

    申请日：2006-02-22

    申请人： D. Bennett ,  Elizabeth Cauchon ,  Dominique Fink ,  Brigette Grouix ,  Ariane Hsia ,  Pamela Danagher ,  Joseph Zimmermann

    发明人： D. Bennett ,  Elizabeth Cauchon ,  Dominique Fink ,  Brigette Grouix ,  Ariane Hsia ,  Pamela Danagher ,  Joseph Zimmermann

    IPC分类号： A61K38/47

    CPC分类号： A61K38/51 ,  A61K47/50 ,  C07K2319/00 ,  C12N9/88 ,  Y10S424/81

    摘要： Heparinase enzymes can be used as a medical treatment to reduce localized inflammatory responses. Treatment of activated endothelium with heparinase inhibits leukocyte rolling, adhesion and extravasation. Most of the heparin and heparan sulfate on endothelial cell surfaces and in basement membranes is degraded by exposure to heparinase. In addition, immobilized chemokines, which are attached to heparin/heparan sulfate on activated endothelium are solubilized by heparinase digestion. Heparinase can be infused into the vascular system to inhibit accumulation of leukocytes in inflamed tissue and decrease damage resulting from localized inflammations. Targeting of heparinase to activated endothelium can be accomplished through localized administration and/or use of genetically engineered heparinase containing endothelium ligand-binding domains.

    摘要翻译： 肝素酶可用作治疗局部炎症反应的药物治疗。 用肝素酶处理激活的内皮抑制白细胞滚动，粘连和外渗。 内皮细胞表面和基底膜中的大部分肝素和硫酸乙酰肝素通过暴露于肝素酶而降解。 此外，通过肝素酶消化可溶解附着于活化内皮上的肝素/硫酸乙酰肝素的固定化趋化因子。 肝素酶可以输注到血管系统中，以抑制发炎组织中白细胞的积累，并减少由局部炎症引起的损伤。 肝素酶对活化内皮的靶向可以通过局部施用和/或使用含有内皮配体结合结构域的转基因肝素酶来实现。

    Apparatus and Process for Light Olefin Recovery
    9.
    发明申请
    Apparatus and Process for Light Olefin Recovery 有权
    轻质烯烃回收装置与工艺

    公开(公告)号：US20070264176A1

    公开(公告)日：2007-11-15

    申请号：US11828433

    申请日：2007-07-26

    申请人： Douglas Stewart ,  Joseph Zimmermann ,  Angelo Furfaro ,  Bipin Vora

    发明人： Douglas Stewart ,  Joseph Zimmermann ,  Angelo Furfaro ,  Bipin Vora

    IPC分类号： B01J7/02

    CPC分类号： C07C4/06 ,  C07C7/163 ,  C10G2400/20 ,  Y02P30/42 ,  C07C11/04 ,  C07C11/06 ,  C07C11/02

    摘要： The present invention relates to a process and apparatus for the production of light olefins comprising olefins having from 2 to 3 carbon atoms per molecule from a feedstock containing heavier olefins. An intermediate cut from a fractionation column is used as olefinic feed to an olefin cracking process preferably after undergoing selective hydrogenation of diolefins. In one embodiment, a liquid side draw from a fractionation column is selectively hydrogenated and then returned to the fractionation column from which a vapor side draw containing olefins is cracked in the olefin cracking reactor.

    摘要翻译： 本发明涉及一种用于生产轻质烯烃的方法和设备，其包含每分子含有2至3个碳原子的烯烃，其含有较重的烯烃的原料。 来自分馏塔的中间馏分用作烯烃裂化方法的烯烃进料，优选在进行二烯烃的选择性氢化之后。 在一个实施方案中，从分馏塔抽出的液体侧被选择性氢化，然后返回到在烯烃裂解反应器中含有烯烃的蒸汽侧馏分从其分馏的分馏塔。