Kinetics and inhibition study of tyrosinase by pressure mediated microanalysis | |
Department | 中科院西北特色植物资源化学重点实验室/甘肃省天然药物重点实验室 |
Liu DM(刘冬梅)1,2; Yang JL(杨军丽)1; Ha W(哈伟)1; Chen J(陈娟)1; Shi YP(师彦平)1; Chen J(陈娟); Shi YP(师彦平) | |
2017 | |
Source Publication | Analytical Biochemistry |
ISSN | 0003-2697 |
Volume | 525Pages:54-59 |
Abstract | In the present study, pressure mediated microanalysis (PMMA), a fast, convenient and efficient capillary electrophoresis (CE) method was developed for studying enzyme kinetics of tyrosinase and inhibition kinetics of kojic acid, a model inhibitor of tyrosinase. The enzymatic reaction conditions and CE conditions were optimized in order to obtain high enzyme activity and short analysis time. By PMMA, only the product could be detected at 475 nm, and no voltage was applied to separate the product from the reaction mixture thus greatly simplifying the optimization procedure. The spectrophotometric assay and electrophoretically mediated microanalysis (EMMA) were also performed to validate the developed method. With the present method, the Michaelis-Menten constant (Km) was calculated to be 1.347 mM for tyrosinase. The inhibition constant of kojic acid to free tyrosinase (KI) and kojic acid to tyrosinase/L-DOPA complex (KIS) were calculated to be 36.64 and 74.35 μM, respectively, and the half-maximal inhibitory concentration (IC50) was determined to be 46.64 μM for kojic acid. The developed method is fast and convenient for studying enzyme kinetics, inhibition kinetics and further screening enzyme inhibitors. |
Keyword | Tyrosinase Enzyme Kinetics Study Inhibition Study Kojic Acid Pressure Mediated Microanalysis |
Subject Area | 分析化学与药物化学 |
DOI | 10.1016/j.ab.2017.02.020 |
Funding Organization | the National Natural Science Foundation of China (Nos. 21375136;21575150);the Scholar Program of West Light Project, Chinese Academy of Sciences (CAS);the top priority program of “One-Three-Five” Strategic Planning of Lanzhou Institute of Chemical Physics, CAS |
Indexed By | SCI |
If | 2.334 |
Language | 英语 |
Funding Project | 药物化学成分研究组 |
compositor | 第一作者单位 |
Citation statistics | |
Document Type | 期刊论文 |
Identifier | http://ir.licp.cn/handle/362003/21780 |
Collection | 中科院西北特色植物资源化学重点实验室/甘肃省天然药物重点实验室 |
Corresponding Author | Chen J(陈娟); Shi YP(师彦平) |
Affiliation | 1.Chinese Acad Sci, Lanzhou Inst Chem Phys, Key Lab Nat Med Gansu Prov, Key Lab Chem Northwestern Plant Resources, Lanzhou 730000, Peoples R China 2.Chinese Acad Sci, Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
Recommended Citation GB/T 7714 | Liu DM,Yang JL,Ha W,et al. Kinetics and inhibition study of tyrosinase by pressure mediated microanalysis[J]. Analytical Biochemistry,2017,525:54-59. |
APA | Liu DM.,Yang JL.,Ha W.,Chen J.,Shi YP.,...&师彦平.(2017).Kinetics and inhibition study of tyrosinase by pressure mediated microanalysis.Analytical Biochemistry,525,54-59. |
MLA | Liu DM,et al."Kinetics and inhibition study of tyrosinase by pressure mediated microanalysis".Analytical Biochemistry 525(2017):54-59. |
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