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Article ; Online: Itaconic acid degradation in

Hossain, Abeer H / Ter Beek, Alexander / Punt, Peter J

2019 Volume 6, Page(s) 1

Abstract: Background: Itaconic acid (IA), a C5-dicarboxylic acid, has previously been identified as one of the top twelve biochemicals that can be produced by biotechnological means. IA is naturally produced by : Results: Transcriptome analysis has led to the ... ...

Abstract	Background: Itaconic acid (IA), a C5-dicarboxylic acid, has previously been identified as one of the top twelve biochemicals that can be produced by biotechnological means. IA is naturally produced by Results: Transcriptome analysis has led to the identification of two novel and previously unknown IA bioconversion pathways in Conclusion: Heterologous IA production in
Language	English
Publishing date	2019-01-04
Publishing country	England
Document type	Journal Article
ZDB-ID	2806612-1
ISSN	2054-3085 ; 2054-3085
ISSN (online)	2054-3085
ISSN	2054-3085
DOI	10.1186/s40694-018-0062-5
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Article ; Online: Identification of novel citramalate biosynthesis pathways in

Hossain, Abeer H / Hendrikx, Aiko / Punt, Peter J

Fungal biology and biotechnology

2019 Volume 6, Page(s) 19

Abstract: Background: The filamentous fungus : Results: IA biodegradation manifests in diminishing titers of IA and the occurrence of an unidentified compound in the HPLC profile. Based on published results on the IA biodegradation pathway, we hypothesized ... ...

Abstract	Background: The filamentous fungus Results: IA biodegradation manifests in diminishing titers of IA and the occurrence of an unidentified compound in the HPLC profile. Based on published results on the IA biodegradation pathway, we hypothesized that the final product of IA biodegradation in Conclusion: In this study, we have shown that the end product of IA biodegradation pathway in
Language	English
Publishing date	2019-11-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2806612-1
ISSN	2054-3085 ; 2054-3085
ISSN (online)	2054-3085
ISSN	2054-3085
DOI	10.1186/s40694-019-0084-7
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Article ; Online: Microbial itaconic acid production from starchy food waste by newly isolated thermotolerant Aspergillus terreus strain.

Narisetty, Vivek / Prabhu, Ashish A / Al-Jaradah, Khalid / Gopaliya, Deeksha / Hossain, Abeer H / Kumar Khare, Sunil / Punt, Peter J / Kumar, Vinod

Bioresource technology

2021 Volume 337, Page(s) 125426

Abstract: In the present study, we have explored the potential of newly isolated Aspergillus terreus BD strain, which can accumulate itaconic acid (IA) at higher temperature. The shake flask cultivation of thermotolerant strain with medium optimized using Box- ... ...

Abstract	In the present study, we have explored the potential of newly isolated Aspergillus terreus BD strain, which can accumulate itaconic acid (IA) at higher temperature. The shake flask cultivation of thermotolerant strain with medium optimized using Box-Behnken Design at 45 °C resulted in IA accumulation of 28.9 g/L with yield of 0.27 g/g. The enzymatic saccharification of the synthetic food waste (SFW) consisting of potatoes, rice & noodles were optimized using Taguchi method of orthogonal array to maximize the release of fermentable sugar. The maximum glucose release of 0.60 g/g was achieved with 10% biomass loading, 5% enzyme concentration, pH 5.5 and temperature 60
MeSH term(s)	Aspergillus ; Fermentation ; Food ; Refuse Disposal ; Succinates
Chemical Substances	Succinates ; itaconic acid (Q4516562YH)
Language	English
Publishing date	2021-06-17
Publishing country	England
Document type	Journal Article
ZDB-ID	1065195-0
ISSN	1873-2976 ; 0960-8524
ISSN (online)	1873-2976
ISSN	0960-8524
DOI	10.1016/j.biortech.2021.125426
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Article: Microbial itaconic acid production from starchy food waste by newly isolated thermotolerant Aspergillus terreus strain

Narisetty, Vivek / Prabhu, Ashish A / Al-Jaradah, Khalid / Gopaliya, Deeksha / Hossain, Abeer H / Kumar Khare, Sunil / Punt, Peter J / Kumar, Vinod

Bioresource technology. 2021 Oct., v. 337

2021

Abstract	In the present study, we have explored the potential of newly isolated Aspergillus terreus BD strain, which can accumulate itaconic acid (IA) at higher temperature. The shake flask cultivation of thermotolerant strain with medium optimized using Box-Behnken Design at 45 °C resulted in IA accumulation of 28.9 g/L with yield of 0.27 g/g. The enzymatic saccharification of the synthetic food waste (SFW) consisting of potatoes, rice & noodles were optimized using Taguchi method of orthogonal array to maximize the release of fermentable sugar. The maximum glucose release of 0.60 g/g was achieved with 10% biomass loading, 5% enzyme concentration, pH 5.5 and temperature 60 ⁰C. The sugars obtained from SFW was integrated with IA production and maximum IA titer achieved with SFW hydrolysate during bioreactor cultivation was 41.1 g/L with conversion yield of 0.27 g/g while with pure glucose IA titer and yield were 44.7 g/L and 0.30 g/g, respectively.
Keywords	Aspergillus terreus ; Taguchi method ; biomass ; bioreactors ; enzymes ; experimental design ; food waste ; glucose ; heat tolerance ; hydrolysates ; itaconic acid ; liquid state fermentation ; pH ; rice ; saccharification ; temperature
Language	English
Dates of publication	2021-10
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	1065195-0
ISSN	1873-2976 ; 0960-8524
ISSN (online)	1873-2976
ISSN	0960-8524
DOI	10.1016/j.biortech.2021.125426
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Article ; Online: Disruption of a putative mitochondrial oxaloacetate shuttle protein in Aspergillus carbonarius results in secretion of malic acid at the expense of citric acid production.

Yang, Lei / Linde, Tore / Hossain, Abeer H / Lübeck, Mette / Punt, Peter J / Lübeck, Peter S

BMC biotechnology

2019 Volume 19, Issue 1, Page(s) 72

Abstract: Background: In filamentous fungi, transport of organic acids across the mitochondrial membrane is facilitated by active transport via shuttle proteins. These transporters may transfer different organic acids across the membrane while taking others the ... ...

Abstract	Background: In filamentous fungi, transport of organic acids across the mitochondrial membrane is facilitated by active transport via shuttle proteins. These transporters may transfer different organic acids across the membrane while taking others the opposite direction. In Aspergillus niger, accumulation of malate in the cytosol can trigger production of citric acid via the exchange of malate and citrate across the mitochondrial membrane. Several mitochondrial organic acid transporters were recently studied in A. niger showing their effects on organic acid production. Results: In this work, we studied another citric acid producing fungus, Aspergillus carbonarius, and identified by genome-mining a putative mitochondrial transporter MtpA, which was not previously studied, that might be involved in production of citric acid. This gene named mtpA encoding a putative oxaloacetate transport protein was expressed constitutively in A. carbonarius based on transcription analysis. To study its role in organic acid production, we disrupted the gene and analyzed its effects on production of citric acid and other organic acids, such as malic acid. In total, 6 transformants with gene mtpA disrupted were obtained and they showed secretion of malic acid at the expense of citric acid production. Conclusion: A putative oxaloacetate transporter gene which is potentially involved in organic acid production by A. carbonarius was identified and further investigated on its effects on production of citric acid and malic acid. The mtpA knockout strains obtained produced less citric acid and more malic acid than the wild type, in agreement with our original hypothesis. More extensive studies should be conducted in order to further reveal the mechanism of organic acid transport as mediated by the MtpA transporter.
MeSH term(s)	Aspergillus/metabolism ; Citric Acid/metabolism ; Malates/metabolism ; Metabolic Engineering/methods ; Mitochondrial Proteins/metabolism ; Oxaloacetates/metabolism
Chemical Substances	Malates ; Mitochondrial Proteins ; Oxaloacetates ; Citric Acid (2968PHW8QP)
Language	English
Publishing date	2019-11-04
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1472-6750
ISSN (online)	1472-6750
DOI	10.1186/s12896-019-0572-0
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Article: Disruption of a putative mitochondrial oxaloacetate shuttle protein in Aspergillus carbonarius results in secretion of malic acid at the expense of citric acid production

Yang, Lei / Linde, Tore / Hossain, Abeer H / Lübeck, Mette / Punt, Peter J / Lübeck, Peter S

BMC biotechnology. 2019 Dec., v. 19, no. 1

2019

Abstract: BACKGROUND: In filamentous fungi, transport of organic acids across the mitochondrial membrane is facilitated by active transport via shuttle proteins. These transporters may transfer different organic acids across the membrane while taking others the ... ...

Abstract	BACKGROUND: In filamentous fungi, transport of organic acids across the mitochondrial membrane is facilitated by active transport via shuttle proteins. These transporters may transfer different organic acids across the membrane while taking others the opposite direction. In Aspergillus niger, accumulation of malate in the cytosol can trigger production of citric acid via the exchange of malate and citrate across the mitochondrial membrane. Several mitochondrial organic acid transporters were recently studied in A. niger showing their effects on organic acid production. RESULTS: In this work, we studied another citric acid producing fungus, Aspergillus carbonarius, and identified by genome-mining a putative mitochondrial transporter MtpA, which was not previously studied, that might be involved in production of citric acid. This gene named mtpA encoding a putative oxaloacetate transport protein was expressed constitutively in A. carbonarius based on transcription analysis. To study its role in organic acid production, we disrupted the gene and analyzed its effects on production of citric acid and other organic acids, such as malic acid. In total, 6 transformants with gene mtpA disrupted were obtained and they showed secretion of malic acid at the expense of citric acid production. CONCLUSION: A putative oxaloacetate transporter gene which is potentially involved in organic acid production by A. carbonarius was identified and further investigated on its effects on production of citric acid and malic acid. The mtpA knockout strains obtained produced less citric acid and more malic acid than the wild type, in agreement with our original hypothesis. More extensive studies should be conducted in order to further reveal the mechanism of organic acid transport as mediated by the MtpA transporter.
Keywords	Aspergillus carbonarius ; Aspergillus niger ; active transport ; citrates ; citric acid ; cytosol ; fungi ; genes ; genome mining ; malates ; malic acid ; mitochondria ; mitochondrial membrane ; oxaloacetic acid ; secretion ; transcription (genetics) ; transporters
Language	English
Dates of publication	2019-12
Size	p. 72.
Publishing place	BioMed Central
Document type	Article
ISSN	1472-6750
DOI	10.1186/s12896-019-0572-0
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Article: Metabolic engineering with ATP-citrate lyase and nitrogen source supplementation improves itaconic acid production in

Hossain, Abeer H / van Gerven, Roy / Overkamp, Karin M / Lübeck, Peter S / Taşpınar, Hatice / Türker, Mustafa / Punt, Peter J

Biotechnology for biofuels

2019 Volume 12, Page(s) 233

Abstract: Background: Bio-based production of organic acids promises to be an attractive alternative for the chemicals industry to substitute petrochemicals as building-block chemicals. In recent years, itaconic acid (IA, methylenesuccinic acid) has been ... ...

Abstract	Background: Bio-based production of organic acids promises to be an attractive alternative for the chemicals industry to substitute petrochemicals as building-block chemicals. In recent years, itaconic acid (IA, methylenesuccinic acid) has been established as a sustainable building-block chemical for the manufacture of various products such as synthetic resins, coatings, and biofuels. The natural IA producer Results: To increase the cytosolic acetyl-CoA pool, we have overexpressed genes Conclusion: Ultimately, this study shows that metabolic engineering of ACL in our rewired IA biosynthesis pathway leads to improved IA production in
Language	English
Publishing date	2019-09-30
Publishing country	England
Document type	Journal Article
ZDB-ID	2421351-2
ISSN	1754-6834
ISSN	1754-6834
DOI	10.1186/s13068-019-1577-6
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Article ; Online: The interplay between transport and metabolism in fungal itaconic acid production.

Hosseinpour Tehrani, Hamed / Geiser, Elena / Engel, Meike / Hartmann, Sandra K / Hossain, Abeer H / Punt, Peter J / Blank, Lars M / Wierckx, Nick

Fungal genetics and biology : FG & B

2019 Volume 125, Page(s) 45–52

Abstract: Besides enzymatic conversions, many eukaryotic metabolic pathways also involve transport proteins that shuttle molecules between subcellular compartments, or into the extracellular space. Fungal itaconate production involves two such transport steps, ... ...

Abstract	Besides enzymatic conversions, many eukaryotic metabolic pathways also involve transport proteins that shuttle molecules between subcellular compartments, or into the extracellular space. Fungal itaconate production involves two such transport steps, involving an itaconate transport protein (Itp), and a mitochondrial tricarboxylate transporter (Mtt). The filamentous ascomycete Aspergillus terreus and the unicellular basidiomycete Ustilago maydis both produce itaconate, but do so via very different molecular pathways, and under very different cultivation conditions. In contrast, the transport proteins of these two strains are assumed to have a similar function. This study aims to investigate the roles of both the extracellular and mitochondrial transporters from these two organisms by expressing them in the corresponding U. maydis knockouts and monitoring the extracellular product concentrations. Both transporters from A. terreus complemented their corresponding U. maydis knockouts in mediating itaconate production. Surprisingly, complementation with At_MfsA from A. terreus led to a partial switch from itaconate to (S)-2-hydroxyparaconate secretion. Apparently, the export protein from A. terreus has a higher affinity for (S)-2-hydroxyparaconate than for itaconate, even though this species is classically regarded as an itaconate producer. Complementation with At_MttA increased itaconate production by 2.3-fold compared to complementation with Um_Mtt1, indicating that the mitochondrial carrier from A. terreus supports a higher metabolic flux of itaconic acid precursors than its U. maydis counterpart. The biochemical implications of these differences are discussed in the context of the biotechnological application in U. maydis and A. terreus for the production of itaconate and (S)-2-hydroxyparaconate.
MeSH term(s)	4-Butyrolactone/analogs & derivatives ; 4-Butyrolactone/biosynthesis ; 4-Butyrolactone/genetics ; Aspergillus/genetics ; Aspergillus/metabolism ; Carrier Proteins/genetics ; Carrier Proteins/metabolism ; Cloning, Molecular ; Fungal Proteins/genetics ; Gene Expression Regulation, Fungal ; Gene Knockout Techniques ; Metabolic Networks and Pathways/genetics ; Mitochondria/genetics ; Succinates/metabolism ; Ustilago/genetics ; Ustilago/metabolism
Chemical Substances	Carrier Proteins ; Fungal Proteins ; Succinates ; citrate-binding transport protein ; paraconic acid ; 4-Butyrolactone (OL659KIY4X) ; itaconic acid (Q4516562YH)
Language	English
Publishing date	2019-01-28
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1319820-8
ISSN	1096-0937 ; 1087-1845 ; 0147-5975
ISSN (online)	1096-0937
ISSN	1087-1845 ; 0147-5975
DOI	10.1016/j.fgb.2019.01.011
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Article: Metabolic engineering with ATP-citrate lyase and nitrogen source supplementation improves itaconic acid production in Aspergillus niger

Hossain, Abeer H / van Gerven, Roy / Overkamp, Karin M / Lübeck, Peter S / Taşpınar, Hatice / Türker, Mustafa / Punt, Peter J

Biotechnology for biofuels. 2019 Dec., v. 12, no. 1

2019

Abstract: BACKGROUND: Bio-based production of organic acids promises to be an attractive alternative for the chemicals industry to substitute petrochemicals as building-block chemicals. In recent years, itaconic acid (IA, methylenesuccinic acid) has been ... ...

Abstract	BACKGROUND: Bio-based production of organic acids promises to be an attractive alternative for the chemicals industry to substitute petrochemicals as building-block chemicals. In recent years, itaconic acid (IA, methylenesuccinic acid) has been established as a sustainable building-block chemical for the manufacture of various products such as synthetic resins, coatings, and biofuels. The natural IA producer Aspergillus terreus is currently used for industrial IA production; however, the filamentous fungus Aspergillus niger has been suggested to be a more suitable host for this purpose. In our previous report, we communicated the overexpression of a putative cytosolic citrate synthase citB in an A. niger strain carrying the full IA biosynthesis gene cluster from A. terreus, which resulted in the highest final titer reported for A. niger (26.2 g/L IA). In this research, we have attempted to improve this pathway by increasing the cytosolic acetyl-CoA pool. Additionally, we have also performed fermentation optimization by varying the nitrogen source and concentration. RESULTS: To increase the cytosolic acetyl-CoA pool, we have overexpressed genes acl1 and acl2 that together encode for ATP-citrate lyase (ACL). Metabolic engineering of ACL resulted in improved IA production through an apparent increase in glycolytic flux. Strains that overexpress acl12 show an increased yield, titer and productivity in comparison with parental strain CitB#99. Furthermore, IA fermentation conditions were improved by nitrogen supplementation, which resulted in alkalization of the medium and thereby reducing IA-induced weak-acid stress. In turn, the alkalizing effect of nitrogen supplementation enabled an elongated idiophase and allowed final titers up to 42.7 g/L to be reached at a productivity of 0.18 g/L/h and yield of 0.26 g/g in 10-L bioreactors. CONCLUSION: Ultimately, this study shows that metabolic engineering of ACL in our rewired IA biosynthesis pathway leads to improved IA production in A. niger due to an increase in glycolytic flux. Furthermore, IA fermentation conditions were improved by nitrogen supplementation that alleviates IA induced weak-acid stress and extends the idiophase.
Keywords	ATP citrate synthase ; Aspergillus niger ; Aspergillus terreus ; acetyl coenzyme A ; alkalinization ; biofuels ; bioreactors ; biosynthesis ; citrate (si)-synthase ; coatings ; fermentation ; fungi ; gene overexpression ; glycolysis ; manufacturing ; metabolic engineering ; multigene family ; nitrogen ; organic acids and salts ; petrochemicals ; resins
Language	English
Dates of publication	2019-12
Size	p. 233.
Publishing place	BioMed Central
Document type	Article
ZDB-ID	2421351-2
ISSN	1754-6834
ISSN	1754-6834
DOI	10.1186/s13068-019-1577-6
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Article ; Online: Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger.

Hossain, Abeer H / Li, An / Brickwedde, Anja / Wilms, Lars / Caspers, Martien / Overkamp, Karin / Punt, Peter J

Microbial cell factories

2016 Volume 15, Issue 1, Page(s) 130

Abstract: Background: The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the ... ...

Abstract	Background: The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the chemical industry to replace petrochemicals. Itaconic acid (IA) has been identified as one of the top twelve building block chemicals which have high potential to be produced by biotechnological means. The IA biosynthesis cluster (cadA, mttA and mfsA) has been elucidated in its natural producer Aspergillus terreus and transferred to A. niger to enable IA production. Here we report the rewiring of a secondary metabolite pathway towards further improved IA production through the overexpression of a putative cytosolic citrate synthase citB in a A. niger strain carrying the IA biosynthesis cluster. Results: We have previously shown that expression of cadA from A. terreus results in itaconic acid production in A. niger AB1.13, albeit at low levels. This low-level production is boosted fivefold by the overexpression of mttA and mfsA in itaconic acid producing AB1.13 CAD background strains. Controlled batch cultivations with AB1.13 CAD + MFS + MTT strains showed increased production of itaconic acid compared with AB1.13 CAD strain. Moreover, preliminary RNA-Seq analysis of an itaconic acid producing AB1.13 CAD strain has led to the identification of the putative cytosolic citrate synthase citB which was induced in an IA producing strain. We have overexpressed citB in a AB1.13 CAD + MFS + MTT strain and by doing so hypothesize to have targeted itaconic acid production to the cytosolic compartment. By overexpressing citB in AB1.13 CAD + MFS + MTT strains in controlled batch cultivations we have achieved highly increased titers of up to 26.2 g/L IA with a productivity of 0.35 g/L/h while no CA was produced. Conclusions: Expression of the IA biosynthesis cluster in Aspergillus niger AB1.13 strain enables IA production. Moreover, in the AB1.13 CAD strain IA production resulted in overexpression of a putative cytosolic citrate synthase citB. Upon overexpression of citB we have achieved titers of up to 26.2 g/L IA with a productivity of 0.35 g/L/h in controlled batch cultivations. By overexpressing citB we have also diminished side product formation and optimized the production pathway towards IA.
MeSH term(s)	Aspergillus/enzymology ; Aspergillus/genetics ; Aspergillus niger/genetics ; Aspergillus niger/metabolism ; Citrate (si)-Synthase/genetics ; Citrate (si)-Synthase/metabolism ; Cloning, Molecular ; Fermentation ; Gene Transfer Techniques ; Metabolic Networks and Pathways/genetics ; Succinates/metabolism
Chemical Substances	Succinates ; Citrate (si)-Synthase (EC 2.3.3.1) ; itaconic acid (Q4516562YH)
Language	English
Publishing date	2016-07-28
Publishing country	England
Document type	Journal Article
ISSN	1475-2859
ISSN (online)	1475-2859
DOI	10.1186/s12934-016-0527-2
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