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1. 发明授权

US06512098B2 Chemically modified proteins with a carbohydrate moiety 有权
公开(公告)号：US06512098B2
公开(公告)日：2003-01-28
申请号：US09347029
申请日：1999-07-02
申请人： J. Bryan Jones , Benjamin G. Davis
发明人： J. Bryan Jones , Benjamin G. Davis
IPC分类号： C07K100
CPC分类号： B82Y5/00 , C12N9/54
摘要： The present invention relates to a chemically modified mutant protein including a cysteine residue substituted for a residue other than cysteine in a precursor protein, the substituted cysteine residue being subsequently modified by reacting the cysteine residue with a glycosylated thiosulfonate. Also, a method of producing the chemically modified mutant protein is provided. The present invention also relates to a glycosylated methanethiosulfonate. Another aspect of the present invention is a method of modifying the functional characteristics of a protein including providing a protein and reacting the protein with a glycosylated methanethiosulfonate reagent under conditions effective to produce a glycoprotein with altered functional characteristics as compared to the protein. In addition, the present invention relates to methods of determining the structure-function relationships of chemically modified mutant proteins.

2. 发明授权

US06930180B2 Chemically modified proteins with a carbohydrate moiety 失效
标题翻译：具有碳水化合物部分的化学修饰的蛋白质
公开(公告)号：US06930180B2
公开(公告)日：2005-08-16
申请号：US10062970
申请日：2002-02-01
申请人： J. Bryan Jones , Benjamin G. Davis
发明人： J. Bryan Jones , Benjamin G. Davis
IPC分类号： C12N15/09 , C07K14/00 , C12N9/54 , C12N9/96 , C12Q1/25 , C12Q1/37 , C07G3/00
CPC分类号： B82Y5/00 , C12N9/54
摘要： The present invention relates to a chemically modified mutant protein including a cysteine residue substituted for a residue other than cysteine in a precursor protein, the substituted cysteine residue being subsequently modified by reacting the cysteine residue with a glycosylated thiosulfonate. Also, a method of producing the chemically modified mutant protein is provided. The present invention also relates to a glycosylated methanethiosulfonate. Another aspect of the present invention is a method of modifying the functional characteristics of a protein including providing a protein and reacting the protein with a glycosylated methanethiosulfonate reagent under conditions effective to produce a glycoprotein with altered functional characteristics as compared to the protein. In addition, the present invention relates to methods of determining the structure-function relationships of chemically modified mutant proteins.
摘要翻译：本发明涉及一种化学修饰的突变蛋白，包括在前体蛋白中取代半胱氨酸以外的残基的半胱氨酸残基，随后通过半胱氨酸残基与糖基化硫代磺酸酯反应来修饰取代的半胱氨酸残基。此外，提供了一种生产化学修饰的突变蛋白的方法。本发明还涉及糖基化甲磺酸盐。本发明的另一方面是一种改变蛋白质的功能特征的方法，包括提供蛋白质并使蛋白质与糖基化甲磺酸盐试剂在与蛋白质相比产生具有改变的功能特征的糖蛋白的条件下反应。此外，本发明涉及确定化学修饰的突变蛋白的结构 - 功能关系的方法。

3. 发明授权

US06576454B2 Modified enzymes and their use for peptide synthesis 有权
公开(公告)号：US06576454B2
公开(公告)日：2003-06-10
申请号：US10075895
申请日：2002-02-13
申请人： J. Bryan Jones
发明人： J. Bryan Jones
IPC分类号： C12N956
CPC分类号： C07K1/023 , C07K5/06026 , C07K5/06078 , C07K5/06086 , C07K5/06104 , C11D3/386 , C12N9/54 , Y10S435/832 , Y10S435/836
摘要： The present invention relates to modified enzymes with one or more amino acid residues from an enzyme being replaced by cysteine residues, where at least some of the cysteine residues are modified by replacing thiol hydrogen in the cysteine residue with a thiol side chain to form a modified enzyme, wherein the modified enzyme has high esterase and low amidase activity. Also, a method of producing the modified enzymes is provided. The present invention also relates to a method for using the modified enzymes in peptide synthesis.

4. 发明授权

US06277617B1 Chemically modified enzymes 失效
标题翻译：化学修饰的酶
公开(公告)号：US06277617B1
公开(公告)日：2001-08-21
申请号：US08978509
申请日：1997-11-25
申请人： Richard R. Bott , Thomas P. Graycar , J. Bryan Jones , Colin Mitchinson
发明人： Richard R. Bott , Thomas P. Graycar , J. Bryan Jones , Colin Mitchinson
IPC分类号： A61K3848
CPC分类号： C11D3/38663 , C12N9/00 , C12N9/54
摘要： Modified enzymes are provided in which at least one amino acid, such as asparagine, leucine, methionine or serine, of an enzyme is replaced with a cysteine and the thiol hydrogen is replaced with a substituent group providing a thiol side chain selected from the group consisting of: a) —SR1R2, wherein R1 is an alkyl and R2 is a charged or polar moiety; b) —SR3, wherein R3 is a substituted or unsubstituted phenyl; c) —SR4, wherein R4 is substituted or unsubstituted cyclohexyl; d) —SR5, wherein R5 is C10-C15 alkyl; and e) —SR6 wherein R6 is a C1-6 alkyl. Also, methods of producing the modified enzymes are provided, as well as detergent and feed additives and a composition for the treatment of a textile. A method for using the modified enzymes in organic synthesis is additionally provided. Further, modified enzymes having improved activity, altered pH profile and/or wash performance are provided.
摘要翻译：提供了修饰的酶，其中酶的至少一个氨基酸，例如天冬酰胺，亮氨酸，甲硫氨酸或丝氨酸被半胱氨酸替代，硫醇氢被提供选自以下的硫醇侧链的取代基取代： a）-SR1R2，其中R1是烷基，R2是带电荷或极性部分; b）-SR3，其中R3是取代或未取代的苯基; c）-SR4，其中R4是取代或未取代的环己基; d） -SR 5，其中R 5为C 10 -C 15烷基; 安德）-SR6，其中R6是C1-6烷基。此外，提供了制备修饰酶的方法，以及洗涤剂和饲料添加剂以及用于处理织物的组合物。另外提供了在有机合成中使用修饰的酶的方法。此外，提供了具有改善的活性，改变的pH曲线和/或洗涤性能的修饰的酶。

5. 发明授权

US06395532B1 Modified enzymes and their use for peptide synthesis 有权
公开(公告)号：US06395532B1
公开(公告)日：2002-05-28
申请号：US09234957
申请日：1999-01-21
申请人： J. Bryan Jones
发明人： J. Bryan Jones
IPC分类号： C12N956
CPC分类号： C07K1/023 , C07K5/06026 , C07K5/06078 , C07K5/06086 , C07K5/06104 , C11D3/386 , C12N9/54 , Y10S435/832 , Y10S435/836
摘要： The present invention relates to modified enzymes with one or more amino acid residues from an enzyme being replaced by cysteine residues, where at least some of the cysteine residues are modified by replacing thiol hydrogen in the cysteine residue with a thiol side chain to form a modified enzyme, wherein the modified enzyme has high esterase and low amidase activity. Also, a method of producing the modified enzymes is provided. The present invention also relates to a method for using the modified enzymes in peptide synthesis.

6. 发明授权

US06284512B1 Chemically modified mutant enzymes and methods for producing them, and screening them for amidase and/or esterase activity 有权
标题翻译：化学修饰的突变酶及其制备方法，并筛选其中的酰胺酶和/或酯酶活性
公开(公告)号：US06284512B1
公开(公告)日：2001-09-04
申请号：US09234956
申请日：1999-01-21
申请人： J. Bryan Jones , Erika Plettner
发明人： J. Bryan Jones , Erika Plettner
IPC分类号： C12N954
CPC分类号： C12Y304/21062 , C11D3/386 , C12N9/54
摘要： The present invention relates to a method for screening chemically modified mutant enzymes for amidase and/or esterase activity. This method includes providing a chemically modified mutant enzyme with one or more amino acid residues from an enzyme being replaced by cysteine residues, where at least some of the cysteine residues are modified by replacing thiol hydrogen in the cysteine residues with a thiol side chain, contacting the chemically modified mutant enzyme with a substrate for an amidase and/or a substrate for an esterase, and determining whether the chemically modified mutant enzyme exhibits amidase and/or esterase activity. The present invention also relates to chemically modified mutant enzymes and a method of producing them where one or more amino acid residues from an enzyme are replaced by cysteine residues, and the cysteine residues are modified by replacing at least some of the thiol hydrogen in the cysteine residue with a thiol side chain to form the chemically modified mutant enzyme. The thiol side chain is selected from the group consisting of —SCH2(p-CH3—C6H4), —SCH2(p-OCH3—C6H4), —SCH2(p-CF3—C6H4), and —SCH2(2,4-diNO2—C6H3).
摘要翻译：本发明涉及用于筛选化学修饰的突变酶用于酰胺酶和/或酯酶活性的方法。该方法包括提供化学修饰的突变酶，其具有由半胱氨酸残基取代的酶中的一个或多个氨基酸残基，其中至少一些半胱氨酸残基通过用巯基侧链取代半胱氨酸残基中的硫醇氢而被修饰，化学修饰的突变酶，其具有用于酰胺酶和/或酯酶底物的底物，以及确定化学修饰的突变酶是否显示酰胺酶和/或酯酶活性。本发明还涉及化学修饰的突变酶及其制备方法，其中来自酶的一个或多个氨基酸残基被半胱氨酸残基取代，半胱氨酸残基被替换半胱氨酸中的至少一些巯基氢残基用硫醇侧链形成化学修饰的突变酶。硫醇侧链选自-SCH 2（p-CH 3 -C 6 H 4），-SCH 2（p-OCH 3 -C 6 H 4），-SCH 2（对CF 3 -C 6 H 4）和-SCH 2（2,4-二硝基 -C 6 H 3）。

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