Characterizing MttA as a mitochondrial cis-aconitic acid transporter by metabolic engineering

被引：45
作者：
Steiger, Matthias G. [1 ,2 ,3 ]
Punt, Peter J. [3 ,4 ]
Ram, Arthur F. J. [3 ]
Mattanovich, Diethard [1 ,2 ]
Sauer, Michael [1 ,2 ,5 ]
机构：
[1] ACIB GmbH, Austrian Ctr Ind Biotechnol, Muthgasse 18, Vienna, Austria
[2] BOKU Univ Nat Resources & Life Sci Vienna, Dept Biotechnol, Muthgasse 18, Vienna, Austria
[3] Leiden Univ, Inst Biol, Sylviusweg 72, Leiden, Netherlands
[4] Dutch DNA Biotech, Utrechtseweg 48, Zeist, Netherlands
[5] CD Lab Biotechnol Glycerol, Muthgasse 18, Vienna, Austria
来源：
METABOLIC ENGINEERING | 2016年 / 35卷
关键词：
Itaconic acid; Aspergillus terreus; Trans-aconitic acid; Mitochondrial carrier protein; Aspergillus niger; Doxycycline; Itaconate; Aconitate; ASPERGILLUS-NIGER; ITACONIC ACID; SACCHAROMYCES-CEREVISIAE; FUNCTIONAL-CHARACTERIZATION; DICARBOXYLATE CARRIER; PROTEIN; GENE; IDENTIFICATION; EXPRESSION; ENZYMES;
D O I：
10.1016/j.ymben.2016.02.003
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
The mitochondrial carrier protein MttA is involved in the biosynthesis of itaconic acid in Aspergillus terreus. In this paper, the transport specificity of MttA is analyzed making use of different metabolically engineered Aspergillus niger strains. Furthermore, the mitochondrial localization of this protein is confirmed using fluorescence microscopy. It was found that MttA preferentially transports cis-aconitic acid over citric acid and does not transport itaconic acid. The expression of MttA in selected A. niger strains results in secretion of aconitic acid. MttA can be used in further strain engineering strategies to transport cis-aconitic acid to the cytosol to produce itaconic acid or related metabolites. The microbial production of aconitic acid (9 g/L) is achieved in strains expressing this transport protein. Thus, metabolic engineering can be used for both the in vivo characterization of transport protein function like MttA and to make use of this protein by creating aconitic acid producing strains. (C) 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
引用
收藏
页码：95 / 104
页数：10
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