Kinetic mechanism of fuculose-1-phosphate aldolase from the hyperthermophilic Archaeon Methanococcus jannaschii

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• 作者：Hae-Chul Park^a ; Joon-Shik Park^b ; Jung-Do Choi^c ; Michael Dabrowski^d ; William M. Atkins^d ; Moon-Young Yoon^a ; ^{myyoon@hanyang.ac.kr}
• 关键词：Fuculose-1-phosphate aldolase ; Methanococcus jannaschii ; Product inhibition ; Dead-end inhibition
• 刊名：Enzyme and Microbial Technology
• 出版年：2012
• 期刊代码：26_01410229
• 类别：ce
• 出版时间：5 April, 2012
• 卷：50
• 期：4-5
• 页码：209-214
• 文件大小：481 K

摘要

Fuculose-1-phosphate aldolase (FucA) is a useful biocatalyst with potential applications in chiral synthesis. In this study, the overall kinetic mechanism of FucA from the archaeon Methanococcus jannaschii was studied. The K_m values of dihydroxyacetone phosphate (DHAP) and dl-glyceraldehyde were 0.09 and 0.74 mM, respectively. Dead-end inhibition by trimethyl phosphonoacetate and dl-threose were competitive and uncompetitive with respect to DHAP and dl-glyceraldehyde. Inhibition patterns obtained using reaction products were noncompetitive vs. DHAP and competitive vs. dl-glyceraldehyde. The equilibrium constant was 8.309 脳 10^鈭? M as assessed by varying the [DHAP]/[product] ratio at a fixed dl-glyceraldehyde concentration and by measuring the change in DHAP concentration after equilibrium was reached. This constant is consistent with the K_eq value obtained from ¹³C NMR (15.625 脳 10^鈭? M). The resultant inhibition kinetics may suggest the insights of kinetic mechanism of the FucA catalyzed reaction.