• 中国精品科技期刊
  • 中国高校百佳科技期刊
  • 中国中文核心期刊
  • 中国科学引文数据库核心期刊
Advanced Search
LIU Huan, WU Xuri, CHEN Yijun. Rapid and sensitive high-throughput determination method for alanine-dependent transaminase[J]. Journal of China Pharmaceutical University, 2018, 49(4): 490-495. DOI: 10.11665/j.issn.1000-5048.20180416
Citation: LIU Huan, WU Xuri, CHEN Yijun. Rapid and sensitive high-throughput determination method for alanine-dependent transaminase[J]. Journal of China Pharmaceutical University, 2018, 49(4): 490-495. DOI: 10.11665/j.issn.1000-5048.20180416

Rapid and sensitive high-throughput determination method for alanine-dependent transaminase

More Information
  • A new method for determining transaminase activity based on the color change of the reaction solution was established, by using alanine-dependent transaminase VfTA from Vibrio fluvialis JS17 as the research object coupled with pyruvate oxidase and horseradish peroxidase. After the optimization of the conditions, the linear relationship between VfTA activity units and the absorbance at 400 nm was investigated. This method was also applied to determine the activity of commercial transaminase ATA117. The results showed that the detection limit of transaminase VfTA activity was up to 0. 45 U/mL and the detection limit of ATA117 activity was up to 0. 5 U/mL. The transaminase activity could be quickly judged according to the color depth of the reaction solution.
  • [1]
    Ward J,Wohlgemuth R.High-yield biocatalytic amination reactions in organic synthesis[J].Curr Org Chem,2010,14(17):1915.
    [2]
    Desai A. Sitagliptin manufacture: a compelling tale of green chemistry,process intensification,and industrial asyetric catalysis[J].Angew Chem Int Edit,2011,50(9):1974.
    [3]
    Ran N,Zhao L,Chen Z,et al.Recent applications of biocatalysis in developing green chemistry for chemical synthesis at the industrial scale[J].J Cheminformatics,2008,39(29):361-372.
    [4]
    Ghislieri D,Turner NJ.Biocatalytic approaches to the synthesis of enantiomerically pure chiral amines[J].Top Catal,2014,57(5):284-300.
    [5]
    Busto E,Gotor-Fernández V,Gotor V.Hydrolases in the stereoselective synthesis of N-heterocyclic amines and amino acid derivatives[J].Chem Rev,2011,111(7):3998-4035.
    [6]
    Abrahamson MJ, Wong JW, Boarius AS. The evolution of an amine dehydrogenase biocatalyst for the asyetric production of chiral amines[J].Adv Synth Catal,2013,355(9):1780-1786.
    [7]
    Malik MS,Park ES,Shin JS.ω-Transaminase-catalyzed kinetic resolution of chiral amines using L-threonine as an amino acceptor precursor[J].Green Chem,2012,14(8):2137-2140.
    [8]
    Green AP,Turner NJ,O′Reilly E.Chiral amine synthesis using ω-transaminases:an amine donor that displaces equilibria and enables high-throughput screening[J].Angew Chem Int Edit,2014,53(40):10714-10717.
    [9]
    Shin JS,Yun H,Jang JW,et al.Purification,characterization,and molecular cloning of a novel amine:pyruvate transaminase from Vibrio fluvialis JS17[J].Appl Microbiol Biotech,2003,61:463-471.
    [10]
    Rosche B,Breuer M,Hauer B,et al.Role of pyruvate in enhancing pyruvate decarboxylase stability towards benzaldehyde[J].J Biotechnol,2005,115(1):91-99.
    [11]
    Genz M, Vickers C, van den Bergh T, et al. Alteration of the donor/acceptor spectrum of the(S)-amine transaminase from Vibrio fluvialis[J].Int J Mol Sci,2015,16:26953-26963.
    [12]
    Yun H,Cho BK,Kim BG.et al.Kinetic resolution of(R,S)-sec-butylamine using Omega-transaminase from Vibrio fluvialis JS17 under reduced pressure[J].Biotechnol Bioeng,2004,36(3):773-780.
    [13]
    Sam M,Giyoung S,Minsu S,et al.High throughput screening methods for ω-transaminases[J].Biotechnol Bioeng,2013,18:1-7.
    [14]
    Katrin W,Margit W,Glieder A,et al.A novel multi- enzymatic high throughput assay for transaminase activity[J].Tetrahedron,2012,68:7586-7590.
    [15]
    Wu X,Fei M,Chen Y,et al.Enzymatic synthesis of L-norephedrine by coupling recombinant pyruvate decarboxylase and ω-transaminase[J].Appl Microbiol Biot,2014,98(17):7399-7408.
    [16]
    Fuchs CS,Simon RC,Riethorst W,et al.Synthesis of(R)- or(S)-valinol using ω-transaminases in aqueous and organic media[J].Bioorgan Med Chem,2014,22(20):5558-5562.
    [17]
    Weiß MS,Pavlidis IV,Vickers C,et al.Glycine oxidase based-throughput solid-phase assay for substrate profiling and directed evolution of(R)- and(S)-selective amine transaminases[J].Anal Chem,2014,86(23):11847-11853.
    [18]
    Scheidt T,Land H,Anderson M,et al.Fluorescence-based kinetic assay for high-throughput discovery and engineering of stereoselective ω-transaminases[J].Adv Synth Catal,2015,357(8):1721-1731.
  • Related Articles

    [1]QIN Tingting, HUANG Zhesu, BAI Haijiao. Content determination of bacteriostats in ocular extractives eye drops and the absorption effect of activated carbon on it[J]. Journal of China Pharmaceutical University, 2020, 51(1): 38-43. DOI: 10.11665/j.issn.1000-5048.20200107
    [2]WANG Huan-ge, XIAO Xue, HU Hao, SU Zhi-jian, XIANG Qi, ZHANG Qi-hao, HUANG Ya-dong. Establishment of an in vitro method to measure the activity of human amylin analogue pramlintide[J]. Journal of China Pharmaceutical University, 2012, 43(4): 355-360.
    [3]REN Jie, CHENG Hong, WANG Wei, HU Kun. Synthesis and antitumor activity of novel chrysin derivatives[J]. Journal of China Pharmaceutical University, 2011, 42(3): 206-212.
    [4]Molecular Weight Determination of Scorpion Venom by LC/MS[J]. Journal of China Pharmaceutical University, 2000, (2): 30-32.
    [5]Purificationand Assays for Activities of HIV-1 Integrase[J]. Journal of China Pharmaceutical University, 2000, (1): 72-75.
    [6]Comparison of Two Bioassay Methods to Determine the in vivo Biological Activity of rhEPO[J]. Journal of China Pharmaceutical University, 1998, (1): 76-78.
    [7]Activity Assay of Superoxide Dismutase in Culture Medium of Ginseng Cells[J]. Journal of China Pharmaceutical University, 1996, (11): 687-689.
    [8]Effects of Tauroursodeoxycholic acid(TUDCA) on Activities of Glutamic Pyruvic Transaminase(GPT)[J]. Journal of China Pharmaceutical University, 1995, (4): 239-241.
    [9]Primary Studies on the Immobilization of an Engineering E. coli Strain with High Activity of Aromatic Amino Acid Transaminase[J]. Journal of China Pharmaceutical University, 1994, (5): 316-318.
    [10]Fu Yanfang. Determination of Erythromycin Content in Intestinal Juice Using Color Development Reaction-Spectrophotometry[J]. Journal of China Pharmaceutical University, 1994, (1).

Catalog

    Article views (1185) PDF downloads (1216) Cited by()

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return