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    METHODS FOR GENERATING NOVEL STABILIZED PROTEINS
    1.
    发明申请
    METHODS FOR GENERATING NOVEL STABILIZED PROTEINS 审中-公开
    生成新型稳定蛋白的方法

    公开(公告)号：WO2008121435A3

    公开(公告)日：2009-04-23

    申请号：PCT/US2008052795

    申请日：2008-02-01

    Applicant: CALIFORNIA INST OF TECHN ,  ARNOLD FRANCES H ,  LI YOUGEN

    Inventor： ARNOLD FRANCES H ,  LI YOUGEN

    IPC: C07K14/00

    CPC classification number: C12N9/0079 ,  C12N9/0042

    Abstract: Methods for identifying and producing stabilized chimeric proteins based on cytochrome P540 are disclosed. Included are polypeptides from the CYP102A1, CYP102A2, and CYP102A3 and combinations thereof. Peptide segments, polynucleotides, vectors, host cells, and enzyme extracts are also disclosed.

    Abstract translation: 公开了基于细胞色素P540鉴定和产生稳定的嵌合蛋白的方法。 包括来自CYP102A1，CYP102A2和CYP102A3的多肽及其组合。 还公开了肽段，多核苷酸，载体，宿主细胞和酶提取物。

    IMPROVED CYTOCHROME P450 OXYGENASES
    2.
    发明申请
    IMPROVED CYTOCHROME P450 OXYGENASES 审中-公开
    改进的CYTOCHROME P450氧化酶

    公开(公告)号：WO03008563A9

    公开(公告)日：2003-08-14

    申请号：PCT/US0223340

    申请日：2002-07-22

    Applicant: CALIFORNIA INST OF TECHN ,  FARINAS EDGARDO T ,  GLIEDER ANTON ,  ARNOLD FRANCES H ,  SCHWANEBERG ULRICH

    Inventor： FARINAS EDGARDO T ,  GLIEDER ANTON ,  ARNOLD FRANCES H ,  SCHWANEBERG ULRICH

    IPC: C12N9/02 ,  C12P7/02 ,  C12P7/04 ,  C12P7/16 ,  C12P17/02 ,  C12N

    CPC classification number: C12N9/0071 ,  C12P7/02 ,  C12P7/04 ,  C12P7/16 ,  C12P17/02 ,  Y02E50/10 ,  Y02P20/52

    Abstract: Nucleic acids encoding cytochrome P450 variants are provided. The cytochrome P450 variants of have a higher alkane-oxidation capability, alkene-oxidation capability, and/or a higher organic-solvent resistance than the corresponding wild-type or parent cytochrome P450 enzyme. A preferred wild-type cytochrome P450 is cytochrome P450 BM-3. Preferred cytochrome P450 variants include those having an improved capability to hydroxylate alkanes and epoxidate alkenes comprising less than 8 carbons, and have amino acid substitutions corresponding to V78A, H236Q, and E252G of cytochrome P450 BM-3. Preferred cytochrome P450 variants also include those having an improved hydroxylation activity in solutions comprising co-solvents such as DMSO and THF, and have amino acid substitutions corresponding to T235A, R471A, E494K, and S1024E of cytochrome P450 BM-3.

    Abstract translation: 提供了编码细胞色素P450变体的核酸。 与相应的野生型或亲本细胞色素P450酶相比，细胞色素P450变体具有更高的烷烃氧化能力，烯烃氧化能力和/或更高的有机溶剂抗性。 优选的野生型细胞色素P450是细胞色素P450 BM-3。 优选的细胞色素P450变体包括具有改进的包含少于8个碳的烷烃和环氧化物烯烃的羟化能力并且具有对应于细胞色素P450 BM-3的V78A，H236Q和E252G的氨基酸取代的那些。 优选的细胞色素P450变体还包括在包含共溶剂例如DMSO和THF的溶液中具有改善的羟基化活性并且具有对应于细胞色素P450 BM-3的T235A，R471A，E494K和S1024E的氨基酸取代的那些。

    OXYGENASE ENZYMES AND SCREENING METHOD
    3.
    发明申请
    OXYGENASE ENZYMES AND SCREENING METHOD 审中-公开
    氧化酶和筛选方法

    公开(公告)号：WO9960096A9

    公开(公告)日：2000-02-24

    申请号：PCT/US9911460

    申请日：1999-05-21

    Applicant: CALIFORNIA INST OF TECHN ,  ARNOLD FRANCES H ,  JOO HYUN ,  LIN ZHANGLIN

    Inventor： ARNOLD FRANCES H ,  JOO HYUN ,  LIN ZHANGLIN

    IPC: C12N9/02 ,  C12N9/08 ,  C12N15/10 ,  C12N15/67 ,  C12N15/81 ,  C12Q1/26 ,  C12Q1/28 ,  C12N

    CPC classification number: C12N9/0071 ,  C07K2319/02 ,  C12N9/0065 ,  C12N15/102 ,  C12N15/1034 ,  C12N15/1058 ,  C12N15/1086 ,  C12N15/67 ,  C12N15/815 ,  C12Q1/26 ,  C12Q1/28 ,  G01N2333/795 ,  G01N2333/80 ,  G01N2333/90245

    Abstract: A method for detecting the presence of an oxygenated compound which is produced when a substrate is reacted with an oxygenase for the substrate. The method involves reacting a coupling enzyme with the oxygenated compound to form a polymeric oxygenated compound which is fluorescent or luminescent. Measurement of the fluorescence or luminescence of the polymeric oxygenated compound provides indirect detection of the oxygenated compound produced by reaction of the oxygenase with the substrate. The method is carried out in a whole cell environment wherein the cell is transformed to express both the oxygenase being screened and the coupling enzyme. The method can be used to measure the activity of monooxygenases and dioxygenases on aromatic substrates. The method is amenable to large scale screening of enzyme mutants to isolate those with maximum oxygenase activity.

    Abstract translation: 用于检测当底物与用于底物的加氧酶反应时产生的含氧化合物的存在的方法。 该方法包括使偶联酶与含氧化合物反应以形成荧光或发光的聚合的氧化化合物。 聚合物氧化化合物的荧光或发光的测量提供了通过加氧酶与底物的反应产生的氧合化合物的间接检测。 该方法在整个细胞环境中进行，其中转化细胞以表达被筛选的加氧酶和偶联酶。 该方法可用于测量单加氧酶和双加氧酶在芳香族底物上的活性。 该方法适用于大规模筛选酶突变体以分离具有最大加氧酶活性的那些。

    DEVICES AND METHODS FOR ANALYSIS OF NON-IONIC SOLUTES
    6.
    发明申请
    DEVICES AND METHODS FOR ANALYSIS OF NON-IONIC SOLUTES 审中-公开
    用于分析非离子溶液的装置和方法

    公开(公告)号：WO0010007A9

    公开(公告)日：2000-07-06

    申请号：PCT/US9918615

    申请日：1999-08-16

    Applicant: CALIFORNIA INST OF TECHN ,  ARNOLD FRANCES H ,  MICHAELS ALAN S ,  ZHENG WEIGONG

    Inventor： ARNOLD FRANCES H ,  MICHAELS ALAN S ,  ZHENG WEIGONG

    IPC: G01N27/30 ,  G01N33/487 ,  A61B5/00

    CPC classification number: G01N27/3271

    Abstract: This invention relates to devices and methods for detecting and monitoring the concentration, level or amount of a target molecule in solution. More particularly, the invention is directed to sensors that evaluate changes in conductivity produced by the reaction of a target molecule with a sensor composition. The invention is particularly well suited for determining the concentration of carbohydrate compounds in solution, such as diol compounds, preferably biologically significant cis-diols such as various sugars including glucose. The invention also encompasses sensors for determining the total or relative amount of target molecule in a sample, particularly a biological sample such as blood, urine, or other biological fluids. A laminated sensor of the invention comprises a sensor composition applied to a metal electrode, such as an interdigitated gold electrode on a non-conductive support. The sensor composition and electrode are and preferably encapsulated by a bipolar membrane. In a preferred embodiment, the sensor composition is a blend of a boronic acid polymer and a non-ionogenic hydrogel matrix.

    Abstract translation: 本发明涉及用于检测和监测溶液中靶分子的浓度，水平或量的装置和方法。 更具体地说，本发明涉及评估由目标分子与传感器组合物反应产生的电导率变化的传感器。 本发明特别适用于测定溶液中碳水化合物的浓度，例如二醇化合物，优选生物学上重要的顺式二醇，如各种糖，包括葡萄糖。 本发明还包括用于测定样品中特别是生物样品如血液，尿液或其他生物液体中目标分子的总量或相对量的传感器。 本发明的层压传感器包括施加到金属电极上的传感器组合物，例如非导电支撑体上的叉指金电极。 传感器组合物和电极优选被双极膜封装。 在优选的实施方案中，传感器组合物是硼酸聚合物和非离子水凝胶基质的混合物。

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