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21. 发明申请

US20060024808A1 Covalent tethering of functional groups to proteins and substrates therefor 有权
公开(公告)号：US20060024808A1
公开(公告)日：2006-02-02
申请号：US11194110
申请日：2005-07-29
申请人： Aldis Darzins , Lance Encell , Dieter Klaubert , Georgyi Los , Mark McDougall , Keith Wood , Monika Wood , Chad Zimprich
发明人： Aldis Darzins , Lance Encell , Dieter Klaubert , Georgyi Los , Mark McDougall , Keith Wood , Monika Wood , Chad Zimprich
IPC分类号： C12Q1/48 , C07H21/04 , C12P21/06 , C12N9/14 , C12N15/74
CPC分类号： G01N33/581 , C07K7/06 , C07K16/00 , C07K2317/622 , C07K2319/43 , C07K2319/61 , C09B1/00 , C09B11/24 , C09B23/04 , C09B23/06 , C09B23/105 , C12N9/14 , C12Q1/34 , C12Y308/01005 , G01N33/573 , G01N2333/914 , Y10S530/825
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

22. 发明申请

US20050272114A1 Substrates for covalent tethering to proteins 有权
标题翻译：用于共价连接蛋白质的底物
公开(公告)号：US20050272114A1
公开(公告)日：2005-12-08
申请号：US11006031
申请日：2004-12-06
申请人： Aldis Darzins , Lance Encell , Tonny Johnson , Dieter Klaubert , Georgyi Los , Mark McDougall , Keith Wood , Monika Wood , Chad Zimprich
发明人： Aldis Darzins , Lance Encell , Tonny Johnson , Dieter Klaubert , Georgyi Los , Mark McDougall , Keith Wood , Monika Wood , Chad Zimprich
IPC分类号： C07K14/47 , C12N9/14 , C12Q1/26
CPC分类号： C12N9/14 , B82Y5/00 , B82Y10/00 , B82Y30/00 , C07K2299/00 , C12N9/0069 , C12Q1/26 , C12Q1/34 , C12Q1/66 , C12Y113/12005 , C12Y308/01005
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译：提供任选融合到感兴趣的蛋白质的突变体水解酶。突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定，并且相对于野生型水解酶具有至少两个氨基酸取代野生型水解酶。还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。

23. 发明授权

US6133016A Sphingomonas biodesulfurization catalyst 失效
标题翻译：鞘氨醇单胞菌生物脱硫催化剂
公开(公告)号：US6133016A
公开(公告)日：2000-10-17
申请号：US851089
申请日：1997-05-05
申请人： Aldis Darzins , Gregory T. Mrachko
发明人： Aldis Darzins , Gregory T. Mrachko
IPC分类号： C10G32/00 , C12N1/21 , C12N9/02 , C12N15/52 , C12S1/02 , C12N1/20 , C07H21/04 , C12N15/00
CPC分类号： C12N9/0051 , C10G32/00 , C12N15/52
摘要： The invention relates to a novel microorganism, designated Sphingomonas sp. strain AD109, which is capable of selectively desulfurizing dibenzothiophene. The invention also includes isolated proteins and nucleic acid sequences obtained from this microorganism. In another embodiment, the invention provides a method of using this microorganism or enzyme preparations derived therefrom in the biocatalytic desulfurization of a fossil fuel containing organic sulfur compounds.
摘要翻译：本发明涉及一种命名为鞘氨醇单胞菌属的新型微生物。能够选择性地脱硫二苯并噻吩的菌株AD109。本发明还包括从该微生物获得的分离的蛋白质和核酸序列。在另一个实施方案中，本发明提供了在含有有机硫化合物的化石燃料的生物催化脱硫中使用该微生物或其衍生的酶制剂的方法。

24. 发明授权

US5952208A Dsz gene expression in pseudomonas hosts 失效
标题翻译： Dsz基因在假单胞菌宿主中的表达
公开(公告)号：US5952208A
公开(公告)日：1999-09-14
申请号：US851088
申请日：1997-05-05
申请人： Aldis Darzins , Lei Xi , John D. Childs , Daniel J. Monticello , Charles H. Squires
发明人： Aldis Darzins , Lei Xi , John D. Childs , Daniel J. Monticello , Charles H. Squires
IPC分类号： C10G32/00 , C12N1/21 , C12N9/02 , C12N15/52 , C12S1/02 , C12P7/22 , C12N15/78
CPC分类号： C12N9/0051 , C10G32/00 , C12N15/52
摘要： The present invention provides novel recombinant pseudomonads which contain a heterologous nucleic acid molecule comprising a nucleotide sequence encoding one or more desulfurization enzymes which are components of a biodesulfurization catalyst. The invention also provides a method of desulfurizing a carbonaceous material, such as a fossil fuel, which comprises organosulfur compounds. The method includes the steps of The method comprises the steps of (1) contacting the fossil fuel with an aqueous phase containing a recombinant biocatalyst which is capable of cleaving carbon-sulfur bonds and, optionally, a flavoprotein, thereby forming a fossil fuel and aqueous phase mixture; (2) maintaining the mixture under conditions sufficient for cleavage of the carbon-sulfur bonds of the organosulfur molecules by the biocatalyst, thereby resulting in a fossil fuel having a reduced organic sulfur content; and (3) separating the fossil fuel having a reduced organic sulfur content from the resulting aqueous phase.
摘要翻译：本发明提供新的重组假单胞菌，其含有包含作为生物脱硫催化剂组分的一种或多种脱硫酶的核苷酸序列的异源核酸分子。本发明还提供一种包含有机硫化合物的含碳材料如化石燃料的脱硫方法。该方法包括以下步骤：该方法包括以下步骤：（1）使化石燃料与含有能够裂解碳 - 硫键的重组生物催化剂的水相和任选的黄素蛋白接触，从而形成化石燃料和水性相混合物（2）在足以使生物催化剂裂解有机硫分子的碳 - 硫键的条件下保持混合物，从而导致具有降低的有机硫含量的化石燃料; 和（3）从所得水相中分离具有降低的有机硫含量的化石燃料。

25. 发明申请

US20060141562A1 Mutant E. coli appa phytase enzymes and natural variants thereof, nucleic acids encoding such phytase enzymes, vectors and host cells incorporating same and methods of making and using same 有权
标题翻译：突变大肠杆菌植酸酶及其天然变体，编码这种植酸酶的核酸，与其结合的载体和宿主细胞以及制备和使用它们的方法
公开(公告)号：US20060141562A1
公开(公告)日：2006-06-29
申请号：US10524234
申请日：2003-08-11
申请人： Beat Blattmann , Aldis Darzins , John Davis , Lance Encell , Thomas Morrison , Gregory Mrachko , Volker Schellenberger
发明人： Beat Blattmann , Aldis Darzins , John Davis , Lance Encell , Thomas Morrison , Gregory Mrachko , Volker Schellenberger
IPC分类号： C12P21/06 , C12N9/16 , C07H21/04 , C12N1/21 , C12N15/74
CPC分类号： C12N15/8243 , C12N9/16 , C12N15/8245
摘要： Cells producing mutant phytases having modified activity are provided, as well as the phytases so produced. Also provided are methods of making and producing such phytases and the use of the expressed phytase protein in feed as a supplement.
摘要翻译：提供具有改变活性的产生突变体植酸酶的细胞，以及如此制备的植酸酶。还提供了制备和生产这种植酸酶的方法以及在饲料中使用表达的植酸酶蛋白作为补充物。

26. 发明申请

US20070087400A1 Covalent tethering of functional groups to proteins and substrates therefor 审中-公开
标题翻译：官能团共价结合到蛋白质和底物上
公开(公告)号：US20070087400A1
公开(公告)日：2007-04-19
申请号：US11509796
申请日：2006-08-24
申请人： Aldis Darzins , Lance Encell , Tonny Johnson , Dieter Klaubert , Georgyi Los , Mark McDougall , Keith Wood , Monika Wood , Chad Zimprich
发明人： Aldis Darzins , Lance Encell , Tonny Johnson , Dieter Klaubert , Georgyi Los , Mark McDougall , Keith Wood , Monika Wood , Chad Zimprich
IPC分类号： C12Q1/26 , C12P21/06 , C07H21/04 , C12N9/16 , C12N9/88 , C12N1/21
CPC分类号： C12Q1/34 , B82Y30/00
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译：提供任选融合到感兴趣的蛋白质的突变体水解酶。突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定，并且相对于野生型水解酶具有至少两个氨基酸取代野生型水解酶。还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。

27. 发明授权

US5821088A Use of gram-positive bacteria to express recombinant proteins 失效
标题翻译：使用革兰氏阳性细菌表达重组蛋白
公开(公告)号：US5821088A
公开(公告)日：1998-10-13
申请号：US472244
申请日：1995-06-07
申请人： Aldis Darzins , Stephen Whitehead , Dennis Hruby , Vincent A. Fischetti
发明人： Aldis Darzins , Stephen Whitehead , Dennis Hruby , Vincent A. Fischetti
IPC分类号： C12N15/09 , A61K38/00 , A61K39/00 , C07K14/31 , C07K14/315 , C12N15/62 , C12N15/74 , C12P21/02 , C12R1/46 , C12P21/04
CPC分类号： C07K14/31 , C07K14/315 , C12N15/62 , C12N15/74 , A61K38/00 , A61K39/00 , C07K2319/00 , C07K2319/02 , C07K2319/035 , C07K2319/23 , C07K2319/50 , C07K2319/705 , C07K2319/735
摘要： A novel system for cloning and expression of genes in gram-positive bacteria. The expression system is based on the finding that many gram-positive bacteria sort proteins to their cell surface through cis-acting N-terminal signal sequences and C-terminal anchor regions. In particular, the cell sorting signals of the streptococcal M6 protein, a well-known surface molecule, are used to construct a gram-positive expression system, designated SPEX (Streptococcal Protein Expression). Expression is achieved by cloning the gene of interest into an appropriate SPEX cassette which is then stably introduced into a bacterial host, such as the human commensal Streptococcus gordonii. Depending on the SPEX vector used, recombinant proteins can be anchored to the cell wall prior to release by specific endoproteolytic cleavage or secreted into the culture medium during bacterial growth. The use of host bacteria lacking extracellular proteases should protect secreted proteins from proteolytic degradation. Several expression vectors in this system also produce specifically-tagged recombinant proteins which allows for a one-step purification of the resulting product.
摘要翻译：用于克隆和表达革兰氏阳性菌中基因的新系统。表达系统基于许多革兰氏阳性细菌通过顺式作用的N末端信号序列和C末端锚定区域将蛋白质分解到其细胞表面的发现。特别地，使用众所周知的表面分子的链球菌M6蛋白的细胞分选信号来构建称为SPEX（链球菌蛋白表达）的革兰氏阳性表达系统。通过将感兴趣的基因克隆到适当的SPEX盒中，然后将其稳定地导入细菌宿主例如人共生型链球菌（Streptococcus gordonii）中来实现表达。根据所使用的SPEX载体，重组蛋白质可以在通过特异性内切蛋白酶切割释放之前锚定到细胞壁上，或者在细菌生长过程中分泌到培养基中。缺乏细胞外蛋白酶的宿主细菌的使用应保护分泌的蛋白质免于蛋白水解降解。该系统中的几种表达载体也产生特异性标记的重组蛋白，其允许对所得产物进行一步纯化。

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