A clone-based transcriptomics approach for the identification of genes relevant for itaconic acid production in Aspergillus

Liu, An; Luijk, N. van; Beek, Marloes ter; Caspers, Martien P. M.; Punt, Peter J.; Werf, Mariet van der

doi:10.1016/j.fgb.2011.01.013

Cited by 123 publications

(149 citation statements)

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“…More recently, van der Straat et al (2014) demonstrated that overexpression of a mitochondrial transporter gene and a plasma membrane transporter gene together with the expression of cad in an oxaloacetate hydrolase and glucose oxidase-deficient A. niger resulted in the itaconic acid production of 7.1 g/L at 78 h cultivation after induction of gene expression. In addition to these reports, introduction of cad into Escherichia coli has been reported by Li et al (2011). They demonstrated cad expression under an inducible T7 promoter in E. coli cultivated in Luria-Bertani (LB) medium at 20 C, which resulted in itaconic acid production of 0.08 g/L after overnight cultivation.…”

Section: Full Papermentioning

confidence: 95%

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Production of itaconic acid using metabolically engineered Escherichia coli

Okamoto

Chin

Hiratsuka

et al. 2014

J. Gen. Appl. Microbiol.

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An Escherichia coli system was engineered for the heterologous production of itaconic acid via the expression of cis-aconitate decarboxylase gene (cad), and then maximal itaconic acid levels produced by engineered E. coli were evaluated. Expression of cad in E. coli grown in Luria-Bertani (LB) medium without glucose in a test tube resulted in 0.07 g/L itaconic acid production after 78 h at 20 C. To increase itaconic acid production, E. coli recombinants were constructed by inactivating the isocitrate dehydrogenase gene (icd) and/or the isocitrate lyase gene (aceA). Expression of cad and inactivation of icd resulted in 0.35 g/L itaconic acid production after 78 h, whereas aceA inactivation had no effect on itaconic acid production. The intracellular itaconate concentration in the icd strain was higher than that in the cad-expressing strain without icd inactivation, which suggests that the extracellular secretion of itaconate in E. coli is the rate-determining step during itaconic acid production. pH-stat cultivation using the cad-expressing icd strain in LB medium with 3% glucose in a jar fermenter resulted in 1.71 g/L itaconic acid production after 97 h at 28 C. To further increase itaconic acid production, the aconitase B gene (acnB) was overexpressed in the cad-expressing icd strain. Simultaneous overexpression of acnB with the expression of cad in the icd strain led to 4.34 g/L itaconic acid production after 105 h. Our findings indicate that icd inactivation and acnB overexpression considerably enhance itaconic acid production in cad-expressing E. coli.Key words: aconitase; cis-aconitate decarboxylase; isocitrate dehydrogenase; isocitrate lyase; itaconic acid; metabolic engineering IntroductionOver the past several decades, there has been substantial interest in itaconic acid as a dicarboxyvinyl monomer produced by microbial fermentation of biomass (Kobayashi and Nakamura, 1964;Okabe et al., 2009;Willke and Vorlop, 2001). Owing to its useful properties, itaconic acid is used in the manufacturing of synthetic polymers, such as plastics and resins (Milson and Meers, 1985;Tate, 1981). Graft polymers with an itaconic acid-based main chain and oligolactate side chain have been synthesized and characterized in our laboratory Okada et al., 2012). The polymers consisting of itaconic acid had high biomass content and are therefore thought to contribute to carbon emission reduction.Itaconic acid is industrially produced from sugars, such as glucose, by the fungus Aspergillus terreus via submerged fermentation (Bonnarme et al., 1995). To date, many re searchers have reported increased productivity of a native itaconic acid producer, A. terreus, by using techniques associated with mutation breeding (Yahiro et al., 1995) and fermentation conditions (Okabe et al., 1993;Park et al., 1994;Riscaldati et al., 2000;Träger et al., 1989;Yahiro et al., 1997). Specifically, the spontaneous mutant A. terreus TN-484 is used as a target microorganism for optimizing the fermentation process. In a previous study, a typical yield of 82....

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Section: Full Papermentioning

confidence: 95%

“…In this study, cad expression was performed by the T7 promoter expression system at 20 C according to a previous study by Li et al (2011). Introduction of pLysS considerably improved Cad protein expression levels (data not shown).…”

Section: Itaconic Acid Production By Cad Expressionmentioning

confidence: 99%

Production of itaconic acid using metabolically engineered Escherichia coli

Okamoto

Chin

Hiratsuka

et al. 2014

J. Gen. Appl. Microbiol.

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“…In an effect to improve the application potential of itaconic acid, a great deal of research related to itaconic acid biosynthesis (Bonnarme et al 1995;Jaklitsch et al 1991;Li et al 2011) and its regulation mechanism (Huang et al 2014b;Lin et al 2004;Reddy and Singh 2002;Tevz et al 2010) have been carried out to improve the production efficiency of itaconic acid. In addition to enhancing the biosynthesis pathway, disrupting or weakening the degradation pathway by metabolic engineering is also an effective strategy to promote the accumulation of target products.…”

Section: Introductionmentioning

confidence: 99%

Identification of an itaconic acid degrading pathway in itaconic acid producing Aspergillus terreus

Chen

Huang

Zhong

et al. 2016

Appl Microbiol Biotechnol

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Itaconic acid, one of the most promising and flexible bio-based chemicals, is mainly produced by Aspergillus terreus. Previous studies to improve itaconic acid production in A. terreus through metabolic engineering were mainly focused on its biosynthesis pathway, while the itaconic aciddegrading pathway has largely been ignored. In this study, we used transcriptomic, proteomic, bioinformatic, and in vitro enzymatic analyses to identify three key enzymes, itaconyl-CoA transferase (IctA), itaconyl-CoA hydratase (IchA), and citramalyl-CoA lyase (CclA), that are involved in the catabolic pathway of itaconic acid in A. terreus. In the itaconic acid catabolic pathway in A. terreus, itaconic acid is first converted by IctA into itaconyl-CoA with succinyl-CoA as the CoA donor, and then itaconyl-CoA is hydrated into citramalyl-CoA by IchA. Finally, citramalyl-CoA is cleaved into acetyl-CoA and pyruvate by CclA. Moreover, IctA can also catalyze the reaction between citramalyl-CoA and succinate to generate succinyl-CoA and citramalate. These results, for the first time, identify the three key enzymes, IctA, IchA, and CclA, involved in the itaconic acid degrading pathway in itaconic acid producing A. terreus. The results will facilitate the improvement of itaconic acid production by metabolically engineering the catabolic pathway of itaconic acid in A. terreus.

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“…1 In particular, itaconic acid (IA), a promising vinyl monomer that is industrially produced by Aspergillus terreus, 2 has recently been produced in Escherichia coli, [3][4][5][6] Aspergillus niger, [7][8][9] and Synechocystis sp, 10 which were engineered to express the cisaconitate decarboxylase gene (cad) 11 from A. terreus. Vinyl monomers such as IA are promising building blocks for synthetic polymers, because of their radical polymerizability.…”

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confidence: 99%

“…6 Li et al demonstrated that E. coli cells containing the cad gene produced 0.08 g/L of IA after overnight growth in Luria-Bertani (LB) medium. 3 More recently, Vuoristo et al introduced the citrate synthase and the aconitase genes (acn) from Corynebacterium glutamicum in E. coli expressing cad and inactivating the phosphate acetyltransferase and the lactate dehydrogenase genes. 5 Cells thus modified produced up to 0.69 g/L of IA when grown in mineral medium supplemented with glucose, in an aerobic bioreactor.…”

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confidence: 99%