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  • 1
    Keywords: Life sciences ; Proteins ; Life sciences ; Protein Science ; Springer eBooks
    Description / Table of Contents: Protein Engineering - Past, Present, and Future -- (Semi)rational Protein Design -- Computational Analysis of Protein Tunnels and Channels -- YASARA - A Tool to Obtain Structural Guidance in Biocatalytic Investigations -- A Computational Library Design Protocol for Rapid Improvement of Protein Stability ́€“ FRESCO -- Directed Evolution of Proteins Based on Mutational Scanning -- A Brief Guide to the High-throughput Expression of Directed Evolution Libraries -- Library Growth and Protein Expression: Optimal and Reproducible Microtiter Plate Expression of Recombinant Enzymes in E. coli using MTP Shakers -- Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification -- Functional Analysis of Membrane Proteins Produced by Cell-Free Translation -- Practical Considerations Regarding the Choice of the Best High-throughput Assay -- High-throughput Screening Assays for Lipolytic Enzymes -- Continuous High-throughput Colorimetric Assays for Îł-transaminases -- Colorimetric High-troughput Screening Assays for the Directed Evolution of Fungal Laccases -- Directed Co-evolution of Two Cellulosic Enzymes in Escherichia coli Based on their Synergistic Reactions -- Program-guided Design of High-throughput Enzyme Screening Experiments and Automated Data Analysis/Evaluation -- Solid Phase Agar Plate Assay for Screening Amine Transaminases -- Ultrahigh-throughput Screening of Single-cell Lysates for Directed Evolution and Functional Metagenomics -- Isolation of pH-sensitive Antibody Fragments by Fluorescence-activated Cell Sorting and Yeast Surface Display -- Library Generation and Auxotrophic Selection Assays in Escherichia coli and Thermus thermophiles.℗
    Abstract: This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments.℗
    Pages: XI, 352 p. 82 illus., 63 illus. in color. : online resource.
    ISBN: 9781493973668
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  • 2
    ISSN: 1434-4475
    Keywords: Keywords. Enantioselectivity; Candida antarctica B; Ibuprofen; Lipase; Organic solvent; Vinylester.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary.  The resolution of ibuprofen by transesterification of its corresponding vinylester using lipase B from Candida antarctica is described. Compared to transesterification or hydrolysis of the ibuprofen ethyl ester (E 〈 2, 28–48 h), the reaction with vinylesters occurred significantly faster (1.5–5 h) and with considerably higher enantioselectivity (E = 8–39).
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature biotechnology 22 (2004), S. 1098-1099 
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] The application of enzymes in biocatalysis plays an increasing role in industrial processes, especially in the development of novel approaches for the synthesis of fine chemicals. This requires a priori the availability of a suitable biocatalyst, which traditionally has been identified by methods, ...
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology reviews 26 (2002), S. 0 
    ISSN: 1574-6976
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Esterases (EC 3.1.1.x) represent a diverse group of hydrolases catalyzing the cleavage and formation of ester bonds and are widely distributed in animals, plants and microorganisms. Beside lipases, a considerable number of microbial carboxyl esterases have also been discovered and overexpressed. This review summarizes their properties and classification. Special emphasis is given on their application in organic synthesis for the resolution of racemates and prostereogenic compounds. In addition, recent results for altering their properties by directed evolution are presented.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 0006-3592
    Keywords: directed evolution ; esterase ; epothilon ; Pseudomonas fluorescens ; stereoselectivity ; mutator strain ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The directed evolution of an esterase from Pseudomonas fluorescens using the mutator strain Epicurian coli XL1-Red was investigated. Mutants were assayed for their ability to hydrolyze a sterically hindered 3-hydroxy ester, which can serve as a building block in the synthesis of epothilones. Screening was performed by plating esterase producing colonies derived from mutation cycles onto minimal media agar plates containing indicator substances (neutral red and crystal violet). Esterase-catalyzed hydrolysis of the 3-hydroxy ester (ethyl or glycerol ester) was detected by the formation of a red color due to a pH decrease caused by the released acid. Esterases isolated from positive clones were used in preparative biotransformations of the ethyl ester. One variant containing two mutations (A209D and L181V) stereoselectively hydrolyzed the ethyl ester resulting in 25% ee for the remaining ester. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58: 554-559, 1998.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-6776
    Keywords: Candida antarctica ; lipase ; glucuronic acid ; vitamin C
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The synthesis of n-butanol and cinnamic alcohol esters of glucuronic acid and the esterification of ascorbic acid (vitamin C) with phenylbutyric acid was performed with lipase from Candida antarctica B (CAL-B, SP435) in a mainly solid-phase system. Products were obtained in 15 to 22 % yield. Computer modelling based on the structure of CAL-B was used to elucidate the access of glucuronic acid to the catalytic site of the lipase. A fixation of glucuronic acid via H-bonds to Q157, D134 and H224 during the transition state was observed. © Rapid Science Ltd. 1998
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-6776
    Keywords: antioxidant ; ascorbic acid ; Candida antarctica B lipase ; fatty acid vinyl ester ; vitamin C
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Fatty acid esters of vitamin C (ascorbic acid) where synthesized in a mainly solid-phase system in the presence of small amounts of organic solvent (acetone or t-butanol) catalyzed by immobilized lipase B from Candida antarctica.Highest reaction rates and yields of isolated products were obtained using fatty acid vinyl esters, e.g., 6-O-palmitoyl-l-ascorbic acid was obtained in 91% isolated yield after 48 h. As vitamin C and its esters are very sensitive to oxidation, a mild extraction method for the isolation of reaction products was developed.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-6776
    Keywords: esterase ; Streptomyces diastatochromogenes ; enantioselectivity ; overexpression ; E. coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Highest overexpression of an esterase from Streptomyces diastatochromogenes (EstA) cloned into E. coli was achieved using a rhamnose-inducible promotor. Highest activity (175 U/ml) was observed 5 h after induction. The lyophilized enzyme had a specific activity of 150 U/mg towards p-nitrophenyl acetate and 48 U/mg towards ethyl acetate. EstA was active in a wide range of pH (optimal 7.5) and temperature (optimal 44 °C ) but became unstable above 50 °C. EstA exibited modest enantioselectivity in the hydrolysis of α-phenylethyl acetate.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1573-6776
    Source: Springer Online Journal Archives 1860-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Regioselective synthesis of e.g. 6-O-phenylbutyryl-1-n-butyl-β-D-glucopyranose was achieved in 21 % yield using almond-β-glucosidase and Candida antarctica lipase B. The β-glucosidase reaction was performed in a biphasic (buffer/n-alcohol) system using free and Eupergit CTM-immobilized glucosidase. Immobilized enzyme allowed product formation even at a water content of 1 %. © Rapid Science Ltd. 1998
    Type of Medium: Electronic Resource
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