文献类型：期刊文献

英文题名：NADH-dependent lactate dehydrogenase from Alcaligenes eutrophus H16 reduces 2-oxoadipate to 2-hydroxyadipate

作者：Zhang, Yan[1,2];Ashok, Somasundar[1];Seol, Eunhee[1];Ainala, Satish Kumar[1];Lee, Sun-Gu[1];Madan, Bharat[1];Xu, Jian-He[2];Park, Sunghoon[1,2]

机构：[1]Pusan Natl Univ, Sch Chem & Biomol Engn, Pusan 609735, South Korea;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2014

卷号：19

期号：6

起止页码：1048

外文期刊名：BIOTECHNOLOGY AND BIOPROCESS ENGINEERING

收录：;EI(收录号：20150400442088);WOS:【SCI-EXPANDED(收录号:WOS:000348046500015)】；

基金：This work was financially supported by the Industrial Strategic technology development program, (10045464, Development of biomass-based nylon-6 monomer precursors production technology) funded by the Ministry of Trade, Industry & Energy (MI, Korea). The authors are grateful for the financial assistance provided by the BK21 Plus Program for Advanced Chemical Technology at Pusan National University. In addition, Dr. Ashok Somasundar is grateful for the financial support provided by "2014 Post-Doc. Development Program of Pusan National University".

语种：英文

外文关键词：adipic acid; 2-Oxoadipate; 2-Oxoadipate reductase; 2-Oxoglutarate; Alcaligenes eutrophus H16; lactate dehydrogenase

摘要：Adipic acid is an important monomer for the production of nylon-6,6 polyamide. One novel biological route for the synthesis of adipic acid, which combines the lysine synthetic pathway and glutaconic acid production pathway, has been suggested, but this route has suffered from the lack of an efficient 2-oxoadipate reductase connecting the two pathways or converting 2-oxoadipate to 2-hydroxyadipate. In this study, we report that the lactate dehydrogenase of Alcaligenes eutrophus H16 is a promising catalyst for this reaction. The lactate dehydrogenase gene (Ae-ldhO) was cloned, expressed in Escherichia coli, purified, and characterized. The recombinant enzyme, having a molecular weight of 36.7 kDa, exhibited broad substrate specificity for various 2-oxoacids. NADH was the preferred coenzyme over NADPH for all 2-oxoacids tested. The maximum specific activity of Ae-LdhO on 2-oxoadipate was 454.5 +/- 20.1 U/mg protein at pH 7.0 and 30a"integral. The K (m) values for 2-oxoadipic acid and NADH were 0.32 +/- 0.02 and 0.09 +/- 0.002 mM, respectively. The activity of Ae-LdhO was enhanced in the presence of some metal ions, such as Mg2+, Co2+ or Ni2+, whereas it was completely inhibited by Hg2+, Ag+, Cu2+ and DTT.