Expression and characterization of fumarase (FUMR) from Rhizopus oryzae

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

doi:10.1016/j.funbio.2010.10.003

Expression and characterization of fumarase (FUMR) from Rhizopus oryzae

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

生物与制药工程学院

Nanjing Tech University

科研成果: 期刊稿件 › 文章 › 同行评审

28 引用（Scopus）

摘要

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 Zn²⁺, Fe²⁺, or EDTA were observed on enzyme activity. A slight inhibition of FUMR activity was seen with Mg²⁺, while Ca²⁺ had a small stimulatory effect. The K_m for l-malic acid and fumaric acid were 0.46mM and 3.07mM, 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.

源语言	英语
页（从-至）	49-53
页数	5
期刊	Fungal Biology
卷	115
期	1
DOI	https://doi.org/10.1016/j.funbio.2010.10.003
出版状态	已出版 - 1月 2011

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title = "Expression and characterization of fumarase (FUMR) from Rhizopus oryzae",

abstract = "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 Zn2+, Fe2+, or EDTA were observed on enzyme activity. A slight inhibition of FUMR activity was seen with Mg2+, while Ca2+ had a small stimulatory effect. The Km for l-malic acid and fumaric acid were 0.46mM and 3.07mM, 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.",

keywords = "Characterization, Cloning, Expression, Fumarase, Rhizopus oryzae",

author = "Ping Song and Shuang Li and Yueyue Ding and Qing Xu and He Huang",

year = "2011",

month = jan,

doi = "10.1016/j.funbio.2010.10.003",

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volume = "115",

pages = "49--53",

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TY - JOUR

T1 - Expression and characterization of fumarase (FUMR) from Rhizopus oryzae

AU - Song, Ping

AU - Li, Shuang

AU - Ding, Yueyue

AU - Xu, Qing

AU - Huang, He

PY - 2011/1

Y1 - 2011/1

N2 - 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 Zn2+, Fe2+, or EDTA were observed on enzyme activity. A slight inhibition of FUMR activity was seen with Mg2+, while Ca2+ had a small stimulatory effect. The Km for l-malic acid and fumaric acid were 0.46mM and 3.07mM, 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.

AB - 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 Zn2+, Fe2+, or EDTA were observed on enzyme activity. A slight inhibition of FUMR activity was seen with Mg2+, while Ca2+ had a small stimulatory effect. The Km for l-malic acid and fumaric acid were 0.46mM and 3.07mM, 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.

KW - Characterization

KW - Cloning

KW - Expression

KW - Fumarase

KW - Rhizopus oryzae

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U2 - 10.1016/j.funbio.2010.10.003

DO - 10.1016/j.funbio.2010.10.003

M3 - 文章

C2 - 21215954

AN - SCOPUS:78650810919

SN - 1878-6146

VL - 115

SP - 49

EP - 53

JO - Fungal Biology

JF - Fungal Biology

IS - 1

ER -

Expression and characterization of fumarase (FUMR) from Rhizopus oryzae

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