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1.发明授权Methods and compositions for preparation of selectively protected carbohydrates 有权用于制备选择性保护的碳水化合物的方法和组合物公开(公告)号：US08802401B2
公开(公告)日：2014-08-12
申请号：US12141348
申请日：2008-06-18
申请人： Frances H. Arnold , Chi-Huey Wong , Yuuichi Mitsuda , Michael M. Chen , Clay Bennett , William Greenberg , Jared Crawford Lewis , Sabine Bastian
发明人： Frances H. Arnold , Chi-Huey Wong , Yuuichi Mitsuda , Michael M. Chen , Clay Bennett , William Greenberg , Jared Crawford Lewis , Sabine Bastian
CPC分类号： C12Q1/26 , C12N9/0077 , C12P19/00 , C12P19/02 , G01N2333/90245 , G01N2500/04
摘要： The disclosure relates to engineered P450 polypeptides and use of such polypeptides in chemoenzymatic methods to construct selectively protected carbohydrates, which are useful as building blocks for preparation of carbohydrate derivatives and oligosaccharides
摘要翻译： 本公开涉及工程化P450多肽，并且在化学酶学方法中使用这种多肽来构建选择性保护的碳水化合物，其可用作制备碳水化合物衍生物和寡糖的构建块
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2.发明申请METHODS AND COMPOSITIONS FOR PREPARATION OF SELECTIVELY PROTECTED CARBOHYDRATES 有权制备选择性保护的碳水化合物的方法和组合物公开(公告)号：US20090124515A1
公开(公告)日：2009-05-14
申请号：US12141348
申请日：2008-06-18
申请人： Frances H. Arnold , Chi-Huey Wong , Yuuichi Mitsuda , Michael M. Chen , Clay Bennett , William Greenberg , Jared Crawford Lewis , Sabine Bastian
发明人： Frances H. Arnold , Chi-Huey Wong , Yuuichi Mitsuda , Michael M. Chen , Clay Bennett , William Greenberg , Jared Crawford Lewis , Sabine Bastian
CPC分类号： C12Q1/26 , C12N9/0077 , C12P19/00 , C12P19/02 , G01N2333/90245 , G01N2500/04
摘要： The disclosure relates to engineered P450 polypeptides and use of such polypeptides in chemoenzymatic methods to construct selectively protected carbohydrates, which are useful as building blocks for preparation of carbohydrate derivatives and oligosaccharides
摘要翻译： 本公开涉及工程化P450多肽，并且在化学酶学方法中使用这种多肽来构建选择性保护的碳水化合物，其可用作制备碳水化合物衍生物和寡糖的构建块
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3.发明授权Libraries of optimized cytochrome P450 enzymes and the optimized P450 enzymes 有权优化的细胞色素P450酶和优化的P450酶的文库公开(公告)号：US08603949B2
公开(公告)日：2013-12-10
申请号：US10869813
申请日：2004-06-15
IPC分类号： C40B50/02
CPC分类号： C12N9/0071
摘要： The present disclosure teaches that the recombination of homologous sequences of P450 enzymes, with the aid of SCHEMA to predict a resulting protein structure, is able to generate libraries of chimeras with significant functional diversity. Additionally, the members of these libraries demonstrate superior or unexpected new properties, which correlate with other factors that are observable in the library. Thus, the making of libraries of optimized P450 enzymes, the analysis of libraries to identify an optimized subset, and the optimized chimeras with improved or altered functionalities are all taught in the present disclosure.
摘要翻译： 本公开内容指出，借助于SCHEMA来预测所得蛋白质结构的P450酶的同源序列的重组能够产生具有显着功能多样性的嵌合体库。 此外，这些图书馆的成员展示了优秀或意想不到的新属性，这与库中可观察到的其他因素相关。 因此，在本公开内容中教导了制备优化的P450酶的文库，分析库以鉴定优化的子集，以及具有改进或改变的功能的优化的嵌合体。
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公开(公告)号：US20100304464A1
公开(公告)日：2010-12-02
申请号：US12723597
申请日：2010-03-12
CPC分类号： C12N9/2437 , C12Y302/01091 , C12Y302/01099
摘要： The present disclosure relates to CBH II chimera fusion polypeptides, nucleic acids encoding the polypeptides, and host cells for producing the polypeptides.
摘要翻译： 本公开涉及CBH II嵌合融合多肽，编码多肽的核酸和用于产生多肽的宿主细胞。
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公开(公告)号：US20090142821A1
公开(公告)日：2009-06-04
申请号：US12287681
申请日：2008-10-11
IPC分类号： C12N9/02
CPC分类号： C12N9/0071 , C12Y114/14001
摘要： The invention relates to novel variants of cytochrome P450 oxygenases. These variants have an improved ability to use peroxide as an oxygen donor as compared to the corresponding wild-type enzyme. These variants also have an improved thermostability as compared to the cytochrome P450 BM-3 F87A mutant. Preferred variants include cytochrome P450 BM-3 heme domain mutants having 158V, F87A, H100R, F107L, A135S, M145A/V, N239H, S274T, L3241, I366V, K434E, E442K, and/or V446I amino acid substitutions.
摘要翻译： 本发明涉及细胞色素P450加氧酶的新变体。 与相应的野生型酶相比，这些变体具有改善使用过氧化物作为供氧体的能力。 与细胞色素P450 BM-3 F87A突变体相比，这些变体也具有改善的热稳定性。 优选的变体包括具有158V，F87A，H100R，F107L，A135S，M145A / V，N239H，S274T，L3241，I366V，K434E，E442K和/或V446I氨基酸取代的细胞色素P450 BM-3血红素结构域突变体。
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公开(公告)号：US07465567B2
公开(公告)日：2008-12-16
申请号：US10125640
申请日：2002-04-16
CPC分类号： C12N9/0071 , C12Y114/14001
摘要： The invention relates to novel variants of cytochrome P450 oxygenases. These variants have an improved ability to use peroxide as an oxygen donor as compared to the corresponding wild-type enzyme. These variants also have an improved thermostability as compared to the cytochrome P450 BM-3 F87A mutant. Preferred variants include cytochrome P450 BM-3 heme domain mutants having I58V, F87A, H100R, F107L, A135S, M145A/V, N239H, S274T, L324I, I366V, K434E, E442K, and/or V446I amino acid substitutions.
摘要翻译： 本发明涉及细胞色素P450加氧酶的新变体。 与相应的野生型酶相比，这些变体具有改善使用过氧化物作为供氧体的能力。 与细胞色素P450 BM-3 F87A突变体相比，这些变体也具有改善的热稳定性。 优选的变体包括具有I58V，F87A，H100R，F107L，A135S，M145A / V，N239H，S274T，L324I，I366V，K434E，E442K和/或V446I氨基酸取代的细胞色素P450 BM-3血红素结构域突变体。
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7.发明授权Thermostable peroxide-driven cytochrome P450 oxygenase variants and methods of use 有权耐热过氧化物驱动的细胞色素P450加氧酶变体和使用方法公开(公告)号：US07435570B2
公开(公告)日：2008-10-14
申请号：US10916369
申请日：2004-08-11
CPC分类号： C12N9/0077
摘要： The invention relates to novel variants of cytochrome P450 oxygenases. These variants have at least one mutation improving their ability to use peroxide as an oxygen donor as compared to the corresponding wild-type enzyme. The variants also have at least one mutation improving thermostability as compared to the parent enzyme or corresponding wild-type enzyme. Preferred variants include cytochrome P450 BM-3 heme domain variants having L52I, I58V, F87A, H100R, S106R, F107L, A135S, M145A/V, A184V, N239H, S274T, L324I, V340M, I366V, K434E, E442K, and/or V446I amino acid substitutions.
摘要翻译： 本发明涉及细胞色素P450加氧酶的新变体。 与相应的野生型酶相比，这些变体具有提高其使用过氧化物作为供氧体的能力的至少一个突变。 与亲本酶或相应的野生型酶相比，变体还具有提高热稳定性的至少一个突变。 优选的变体包括具有L52I，I58V，F87A，H100R，S106R，F107L，A135S，M145A / V，A184V，N239H，S274T，L324I，V340M，I366V，K434E，E442K和/或V446I的细胞色素P450 BM-3血红素结构域变体 氨基酸取代。
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公开(公告)号：US07115403B1
公开(公告)日：2006-10-03
申请号：US09571553
申请日：2000-05-16
CPC分类号： C12N9/0006 , C12N15/102
摘要： This invention relates to the expression of improved polynucleotide and polypeptide sequences encoding for eukaryotic enzymes, particularly oxidase enzymes. The enzymes are advantageously produced in conventional or facile expression systems. Various methods for directed evolution of polynucleotide sequences can be used to obtain the improved sequences. The improved characteristics of the polypeptides or proteins generated in this manner include improved expression, enhanced activity toward one or more substrates, and increased thermal stability. In a particular embodiment, the invention relates to improved expression of the galactose oxidase gene and galactose oxidase enzymes. GAO mutants that are highly active and/or thermostable are disclosed.
摘要翻译： 本发明涉及编码真核生物酶，特别是氧化酶的改良的多核苷酸和多肽序列的表达。 酶有利地在常规或简单的表达系统中产生。 可以使用用于多核苷酸序列的定向进化的各种方法来获得改进的序列。 以这种方式产生的多肽或蛋白质的改进特征包括改进的表达，增强对一种或多种底物的活性和增加的热稳定性。 在一个具体实施方案中，本发明涉及半乳糖氧化酶基因和半乳糖氧化酶的改进表达。 公开了高活性和/或热稳定的GAO突变体。
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公开(公告)号：US07098010B1
公开(公告)日：2006-08-29
申请号：US09722602
申请日：2000-11-27
CPC分类号： C12N9/0006 , C12N15/102
摘要： This invention relates to the expression of improved polynucleotide and polypeptide sequences encoding for eukaryotic enzymes, particularly oxidase enzymes. The enzymes are advantagoeusly produced in conventional or facile expression systems. Various methods for directed evolution of polynucleotide sequences can be used to obtain the improved sequences. The improved characteristics of the polypeptides or proteins generated in this manner include improved expression, enhanced activity toward one or more substrates, and increased thermal stability. In a particular embodiment, the invention relates to improved expression of the galactose oxidase gene and galactose oxidase enzymes. GAO mutants that are highly active and/or thermostable are disclosed.
摘要翻译： 本发明涉及编码真核生物酶，特别是氧化酶的改良的多核苷酸和多肽序列的表达。 这些酶在常规或简单的表达系统中有利地产生。 可以使用用于多核苷酸序列的定向进化的各种方法来获得改进的序列。 以这种方式产生的多肽或蛋白质的改进特征包括改进的表达，增强对一种或多种底物的活性和增加的热稳定性。 在一个具体实施方案中，本发明涉及半乳糖氧化酶基因和半乳糖氧化酶的改进表达。 公开了高活性和/或热稳定的GAO突变体。
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10.发明授权Metal chelating lipids which are useful as sensors in fluorometric methods for the detection of metal ions 失效金属螯合脂质，用作检测金属离子的荧光测定方法中的传感器
公开(公告)号：US5837202A
公开(公告)日：1998-11-17
申请号：US712193
申请日：1996-09-11
IPC分类号： C09B57/10 , G01N31/22 , G01N33/533 , G01N21/64 , C07C229/34 , C07D255/02
CPC分类号： G01N31/221 , C09B57/10 , G01N31/22 , G01N33/533
摘要： An fluorescent metal-chelating amphiphile having the structure: ##STR1## wherein A is a hydrophobic fluorophore, X and Y are aliphatic hydrocarbons having from 9 to 25 carbon atoms, B is a hydrophilic spacer, C is a metal chelator, and L is either an ether or ester linkage. The fluorescent metal-chelating amphiphile is combined with a matrix lipid to form lipid-based sensors which provide fluorometric detection of metal ions in liquids.
摘要翻译： 具有以下结构的荧光金属螯合两亲物：其中A是疏水性荧光团，X和Y是具有9至25个碳原子的脂族烃，B是亲水间隔基，C是金属螯合剂， L是醚或酯键。 荧光金属螯合两亲物与基质脂质组合以形成基于脂质的传感器，其提供液体中金属离子的荧光检测。
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