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

COLLEGE OF BIOTECHNOLOGY AND PHARMACEUTICAL ENGINEERING

Nanjing Tech University

Research output: Contribution to journal › Article › peer-review

28 Scopus citations

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 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.

Original language	English
Pages (from-to)	49-53
Number of pages	5
Journal	Fungal Biology
Volume	115
Issue number	1
DOIs	https://doi.org/10.1016/j.funbio.2010.10.003
State	Published - Jan 2011

Keywords

Characterization
Cloning
Expression
Fumarase
Rhizopus oryzae

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.funbio.2010.10.003

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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",

journal = "Fungal Biology",

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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

Abstract

Keywords

UN SDGs

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