Targeting enzymes to the right compartment: Metabolic engineering for itaconic acid production by Aspergillus niger

被引:85
作者
Blumhoff, Marzena L. [1 ,2 ]
Steiger, Matthias G. [1 ,3 ]
Mattanovich, Diethard [1 ,3 ]
Sauer, Michael [1 ,3 ]
机构
[1] BOKU VIBT Univ Nat Resources & Life Sci, Dept Biotechnol, A-1190 Vienna, Austria
[2] Univ Appl Sci FH, Sch Bioengn, Vienna, Austria
[3] Austrian Ctr Ind Biotechnol ACIB GmbH, Vienna, Austria
关键词
Mitochondrial targeting; cis-Aconitate decarboxylase; Aconitase; Microbial organic acid production; AcoA; CadA; CITRATE PRODUCTION; EXPRESSION; GENE; PATHWAYS; DECARBOXYLASE; CONSTRUCTION; FERMENTATION; MORPHOLOGY; NIDULANS; PROTEIN;
D O I
10.1016/j.ymben.2013.05.003
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
Itaconic acid is an unsaturated dicarboxylic acid which has a high potential as a biochemical building block, It can be microbially produced from some Aspergillus species, such as Aspergillus itaconicus and Aspergillus terreus. However, the achieved titers are significantly lower as compared to the citric acid production by A. niger. Heterologous expression of cis-aconitate decarboxylase in A. niger leads to the accumulation of small amounts of itaconic acid. Additional expression of aconitase, the second enzyme metabolically linking citric acid and itaconic acid improves productivity. However, proper organelle targeting of the enzymes appears to be an important point to consider. Here we compare the mitochondrial expression with the cytosolic expression of cis-aconitate decarboxylase or aconitase in A niger. Heterologous expression of both enzymes in the mitochondria doubles the productivity compared to strains which express the enzymes in the cytosol. It is essential to target enzymes to the correct compartment in order to establish a proper flux through a compartmentalized pathway. (C) 2013 Elsevier Inc. All rights reserved.
引用
收藏
页码:26 / 32
页数:7
相关论文
共 39 条
[11]   BIOCHEMISTRY OF THE USTILAGINALES .11. METABOLIC PRODUCTS OF USTILAGO-ZEAE IN SUBMERGED CULTURE [J].
HASKINS, RH ;
THORN, JA ;
BOOTHROYD, B .
CANADIAN JOURNAL OF MICROBIOLOGY, 1955, 1 (09) :749-&
[12]   The cellular target of histatin 5 on Candida albicans is the energized mitochondrion [J].
Helmerhorst, EJ ;
Breeuwer, P ;
van't Hof, W ;
Walgreen-Weterings, E ;
Oomen, LCJM ;
Veerman, ECI ;
Amerongen, AVN ;
Abee, T .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (11) :7286-7291
[13]   INTRACELLULAR LOCATION OF ENZYMES INVOLVED IN CITRATE PRODUCTION BY ASPERGILLUS-NIGER [J].
JAKLITSCH, WM ;
KUBICEK, CP ;
SCRUTTON, MC .
CANADIAN JOURNAL OF MICROBIOLOGY, 1991, 37 (11) :823-827
[14]   THE SUBCELLULAR ORGANIZATION OF ITACONATE BIOSYNTHESIS IN ASPERGILLUS-TERREUS [J].
JAKLITSCH, WM ;
KUBICEK, CP ;
SCRUTTON, MC .
JOURNAL OF GENERAL MICROBIOLOGY, 1991, 137 :533-539
[15]   An efficient fungal RNA-silencing system using the DsRed reporter gene [J].
Janus, Danielle ;
Hoff, Birgit ;
Hofmann, Eckhard ;
Kueck, Ulrich .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (03) :962-970
[16]  
Jore J.P.M., 2011, US Patent, Patent No. [20110124066 A1, 20110124066]
[17]   Cloning and functional characterization of the cis-aconitic acid decarboxylase (CAD) gene from Aspergillus terreus [J].
Kanamasa, Shin ;
Dwiarti, Lies ;
Okabe, Mitsuyasu ;
Park, Enoch Y. .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2008, 80 (02) :223-229
[18]  
Kawamura D., 1981, Japanese Patent, Patent No. [56137893, 56,137,893]
[19]  
Kirimura K, 2002, J BIOSCI BIOENG, V93, P136, DOI 10.1016/S1389-1723(02)80005-6
[20]   Expression analysis of alternative oxidase gene (aox1) with enhanced green fluorescent protein as marker in citric acid-producing Aspergillus niger [J].
Kirimura, Kohtaro ;
Ogawa, Satoshi ;
Hattori, Takasumi ;
Kino, Kuniki .
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 2006, 102 (03) :210-214