Expression and characterization of fumarase (FUMR) from Rhizopus oryzae.

Ping Song, Shuang Li, Yueyue Ding, Qing Xu, He Huang

State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, No. 5 Xinmofan Road, Nanjing 210009, PR China.

Fungal biology 2011 Jan

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Fumarase catalyzes the reversible hydration of fumarate to l-malate in Rhizopus oryzae. A recombinant pET22b-fumR harboring a fumarase gene (fumR) from R. oryzae was constructed for high level expression in E. coli BL21 (DE3). The FUMR activity was optimal at 30°C and pH 7.2. The enzyme was stable below 45°C and at pH 3.0-9.0. No effects of Zn(2+), Fe(2+), or EDTA were observed on enzyme activity. A slight inhibition of FUMR activity was seen with Mg(2+), while Ca(2+) had a small stimulatory effect. The K(m) for l-malic acid and fumaric acid were 0.46 mM and 3.07 mM, respectively. The activity of FUMR catalyzing hydration of fumarate to l-malate was completely inhibited by 2mM fumaric acid. The unique enzymatic properties suggested that overexpression of FUMR could enhance fumaric acid accumulation in R. oryzae. Copyright Â© 2010. Published by Elsevier Ltd.

Citation

Ping Song, Shuang Li, Yueyue Ding, Qing Xu, He Huang. Expression and characterization of fumarase (FUMR) from Rhizopus oryzae. Fungal biology. 2011 Jan;115(1):49-53