scholarly journals An Optimized Ustilago maydis for Itaconic Acid Production at Maximal Theoretical Yield

2020 ◽  
Vol 7 (1) ◽  
pp. 20
Author(s):  
Johanna Becker ◽  
Hamed Hosseinpour Tehrani ◽  
Philipp Ernst ◽  
Lars Mathias Blank ◽  
Nick Wierckx
Keyword(s):  
Itaconic Acid ◽  
Acid Production ◽  
Lipid Production ◽  
Ustilago Maydis ◽  
Product Formation ◽  
Fed Batch ◽  
Theoretical Yield ◽  
Maximal Yield ◽  
Morphology Engineering ◽  
Continuous Feed

Ustilago maydis, a member of the Ustilaginaceae family, is a promising host for the production of several metabolites including itaconic acid. This dicarboxylate has great potential as a bio-based building block in the polymer industry, and is of special interest for pharmaceutical applications. Several itaconate overproducing Ustilago strains have been generated by metabolic and morphology engineering. This yielded stabilized unicellular morphology through fuz7 deletion, reduction of by-product formation through deletion of genes responsible for itaconate oxidation and (glyco)lipid production, and the overexpression of the regulator of the itaconate cluster ria1 and the mitochondrial tricarboxylate transporter encoded by mttA from Aspergillusterreus. In this study, itaconate production was further optimized by consolidating these different optimizations into one strain. The combined modifications resulted in itaconic acid production at theoretical maximal yield, which was achieved under biotechnologically relevant fed-batch fermentations with continuous feed.

scholarly journals An Optimized Ustilago Chassis for Itaconic Acid Production at Theoretical Maximal Yield

2020 ◽  
Author(s):  
J. Becker ◽  
H. Hosseinpour Tehrani ◽  
P. Ernst ◽  
L. M. Blank ◽  
N. Wierckx
Keyword(s):  
Itaconic Acid ◽  
Acid Production ◽  
Aspergillus Terreus ◽  
Lipid Production ◽  
Ustilago Maydis ◽  
Product Formation ◽  
Fed Batch ◽  
Maximal Yield ◽  
Morphology Engineering ◽  
Continuous Feed

Ustilago maydis, member of the Ustilaginaceae family, is a promising host for the production of several metabolites including itaconic acid. This dicarboxylate has great potential as a bio-based building block in the polymer industry, and is of special interest for pharmaceutical applications. Several itaconate overproducing Ustilago strains have been generated by metabolic and morphology engineering. This yielded stabilized unicellular morphology through fuz7 deletion, reduction of by-product formation through deletion of genes responsible for itaconate oxidation and (glyco)lipid production, and the overexpression of the regulator of the itaconate cluster ria1 and the mitochondrial tricarboxylate transporter encoded by mttA from Aspergillus terreus. In this study, itaconate production was further optimized by consolidating these different optimizations into one strain. The combined modifications resulted in itaconic acid production at theoretical maximal yield, which was achieved under biotechnologically relevant fed-batch fermentations with continuous feed.

scholarly journals Consolidated bioprocessing of cellulose to itaconic acid by a co-culture of Trichoderma reesei and Ustilago maydis

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Ivan Schlembach ◽  
Hamed Hosseinpour Tehrani ◽  
Lars M. Blank ◽  
Jochen Büchs ◽  
Nick Wierckx ◽  
...  
Keyword(s):  
Trichoderma Reesei ◽  
Itaconic Acid ◽  
Acid Production ◽  
Ustilago Maydis ◽  
Biofuel Production ◽  
Microbial Consortia ◽  
Substrate Availability ◽  
Fed Batch ◽  
Organic Acid Production ◽  
Acid Yield

Abstract Background Itaconic acid is a bio-derived platform chemical with uses ranging from polymer synthesis to biofuel production. The efficient conversion of cellulosic waste streams into itaconic acid could thus enable the sustainable production of a variety of substitutes for fossil oil based products. However, the realization of such a process is currently hindered by an expensive conversion of cellulose into fermentable sugars. Here, we present the stepwise development of a fully consolidated bioprocess (CBP), which is capable of directly converting recalcitrant cellulose into itaconic acid without the need for separate cellulose hydrolysis including the application of commercial cellulases. The process is based on a synthetic microbial consortium of the cellulase producer Trichoderma reesei and the itaconic acid producing yeast Ustilago maydis. A method for process monitoring was developed to estimate cellulose consumption, itaconic acid formation as well as the actual itaconic acid production yield online during co-cultivation. Results The efficiency of the process was compared to a simultaneous saccharification and fermentation setup (SSF). Because of the additional substrate consumption of T. reesei in the CBP, the itaconic acid yield was significantly lower in the CBP than in the SSF. In order to increase yield and productivity of itaconic acid in the CBP, the population dynamics was manipulated by varying the inoculation delay between T. reesei and U. maydis. Surprisingly, neither inoculation delay nor inoculation density significantly affected the population development or the CBP performance. Instead, the substrate availability was the most important parameter. U. maydis was only able to grow and to produce itaconic acid when the cellulose concentration and thus, the sugar supply rate, was high. Finally, the metabolic processes during fed-batch CBP were analyzed in depth by online respiration measurements. Thereby, substrate availability was again identified as key factor also controlling itaconic acid yield. In summary, an itaconic acid titer of 34 g/L with a total productivity of up to 0.07 g/L/h and a yield of 0.16 g/g could be reached during fed-batch cultivation. Conclusion This study demonstrates the feasibility of consortium-based CBP for itaconic acid production and also lays the fundamentals for the development and improvement of similar microbial consortia for cellulose-based organic acid production.

scholarly journals Consolidated Bioprocessing of Cellulose to Itaconic Acid by a Co-Culture of Trichoderma Reesei and Ustilago Maydis.

2020 ◽  
Author(s):  
Ivan Schlembach ◽  
Hamed Tehrani ◽  
Lars Mathias Blank ◽  
Jochen Büchs ◽  
Nick Wierckx ◽  
...  
Keyword(s):  
Trichoderma Reesei ◽  
Itaconic Acid ◽  
Acid Production ◽  
Ustilago Maydis ◽  
Biofuel Production ◽  
Microbial Consortia ◽  
Substrate Availability ◽  
Fed Batch ◽  
Organic Acid Production ◽  
Acid Yield

Abstract Background: Itaconic acid is a bio-derived platform chemical with uses ranging from polymer synthesis to biofuel production. The efficient conversion of cellulosic waste streams into itaconic acid could thus enable the sustainable production of a variety of substitutes for fossil oil based products. However, the realization of such a process is currently hindered by an expensive conversion of cellulose into fermentable sugars. Here, we present the stepwise development of a fully consolidated bioprocess (CBP), which is capable to directly convert recalcitrant cellulose into itaconic acid without the need for separate cellulose hydrolysis including the application of commercial cellulases. The process is based on a synthetic microbial consortium of the cellulase producer Trichoderma reesei and the itaconic acid producing yeast Ustilago maydis. Results: The efficiency of the process was compared to a simultaneous hydrolysis and fermentation setup (SSF). Because of the additional substrate consumption of T. reesei in the CBP, the itaconic acid yield was significantly lower in the CBP than in the SSF. In order to increase yield and productivity of itaconic acid in the CBP, the population dynamics was manipulated by varying the inoculation delay between T. reesei and U. maydis. Surprisingly, neither inoculation delay nor inoculation density significantly affected the population development or the CBP performance. Instead, the substrate availability was the most important parameter. U. maydis was only able to grow and to produce itaconic acid when the cellulose concentration and thus, the sugar supply rate, was high. Finally, the metabolic processes during fed-batch CBP were analyzed in depth by online respiration measurements. A method was developed to estimate cellulose consumption, itaconic acid formation as well as the actual itaconic acid production yield online. Again, substrate availability was the key factor also controlling itaconic acid yield. In summary, an itaconic acid titer of 34 g/L with a total productivity of up to 0.07 g/L/h and a yield of 0.16 g/g could be reached during fed-batch cultivation. Conclusion: This study demonstrates the feasibility of consortium based CBP for itaconic acid production and also lays the fundament for the development and improvement of similar microbial consortia for cellulose based organic acid production.

scholarly journals Consolidated bioprocessing of cellulose to itaconic acid by a co-culture of Trichoderma reesei and Ustilago maydis.

2020 ◽  
Author(s):  
Ivan Schlembach ◽  
Hamed Tehrani ◽  
Lars Mathias Blank ◽  
Jochen Büchs ◽  
Nick Wierckx ◽  
...  
Keyword(s):  
Trichoderma Reesei ◽  
Itaconic Acid ◽  
Acid Production ◽  
Ustilago Maydis ◽  
Biofuel Production ◽  
Microbial Consortia ◽  
Substrate Availability ◽  
Fed Batch ◽  
Organic Acid Production ◽  
Acid Yield

Abstract Background: Itaconic acid is a bio-derived platform chemical with uses ranging from polymer synthesis to biofuel production. The efficient conversion of cellulosic waste streams into itaconic acid could thus enable the sustainable production of a variety of substitutes for fossil oil based products. However, the realization of such a process is currently hindered by an expensive conversion of cellulose into fermentable sugars. Here, we present the stepwise development of a fully consolidated bioprocess (CBP), which is capable to directly convert recalcitrant cellulose into itaconic acid without the need for separate cellulose hydrolysis including the application of commercial cellulases. The process is based on a synthetic microbial consortium of the cellulase producer Trichoderma reesei and the itaconic acid producing yeast Ustilago maydis. A method for process monitoring was developed to estimate cellulose consumption, itaconic acid formation as well as the actual itaconic acid production yield online during co-cultivation.Results: The efficiency of the process was compared to a simultaneous hydrolysis and fermentation setup (SSF). Because of the additional substrate consumption of T. reesei in the CBP, the itaconic acid yield was significantly lower in the CBP than in the SSF. In order to increase yield and productivity of itaconic acid in the CBP, the population dynamics was manipulated by varying the inoculation delay between T. reesei and U. maydis. Surprisingly, neither inoculation delay nor inoculation density significantly affected the population development or the CBP performance. Instead, the substrate availability was the most important parameter. U. maydis was only able to grow and to produce itaconic acid when the cellulose concentration and thus, the sugar supply rate, was high. Finally, the metabolic processes during fed-batch CBP were analyzed in depth by online respiration measurements. Thereby, substrate availability was again identified as key factor also controlling itaconic acid yield. In summary, an itaconic acid titer of 34 g/L with a total productivity of up to 0.07 g/L/h and a yield of 0.16 g/g could be reached during fed-batch cultivation. Conclusion: This study demonstrates the feasibility of consortium based CBP for itaconic acid production and also lays the fundament for the development and improvement of similar microbial consortia for cellulose based organic acid production.

scholarly journals Consolidated bioprocessing of cellulose to itaconic acid by a co-culture of Trichoderma reesei and Ustilago maydis.

2020 ◽  
Author(s):  
Ivan Schlembach ◽  
Hamed Tehrani ◽  
Lars Mathias Blank ◽  
Jochen Büchs ◽  
Nick Wierckx ◽  
...  
Keyword(s):  
Trichoderma Reesei ◽  
Itaconic Acid ◽  
Acid Production ◽  
Ustilago Maydis ◽  
Biofuel Production ◽  
Microbial Consortia ◽  
Substrate Availability ◽  
Fed Batch ◽  
Organic Acid Production ◽  
Acid Yield

Abstract Background: Itaconic acid is a bio-derived platform chemical with uses ranging from polymer synthesis to biofuel production. The efficient conversion of cellulosic waste streams into itaconic acid could thus enable the sustainable production of a variety of substitutes for fossil oil based products. However, the realization of such a process is currently hindered by an expensive conversion of cellulose into fermentable sugars. Here, we present the stepwise development of a fully consolidated bioprocess (CBP), which is capable to directly convert recalcitrant cellulose into itaconic acid without the need for separate cellulose hydrolysis including the application of commercial cellulases. The process is based on a synthetic microbial consortium of the cellulase producer Trichoderma reesei and the itaconic acid producing yeast Ustilago maydis. A method for process monitoring was developed to estimate cellulose consumption, itaconic acid formation as well as the actual itaconic acid production yield online during co-cultivation. Results: The efficiency of the process was compared to a simultaneous hydrolysis and fermentation setup (SSF). Because of the additional substrate consumption of T. reesei in the CBP, the itaconic acid yield was significantly lower in the CBP than in the SSF. In order to increase yield and productivity of itaconic acid in the CBP, the population dynamics was manipulated by varying the inoculation delay between T. reesei and U. maydis . Surprisingly, neither inoculation delay nor inoculation density significantly affected the population development or the CBP performance. Instead, the substrate availability was the most important parameter. U. maydis was only able to grow and to produce itaconic acid when the cellulose concentration and thus, the sugar supply rate, was high. Finally, the metabolic processes during fed-batch CBP were analyzed in depth by online respiration measurements. Thereby, substrate availability was again identified as the key factor also controlling itaconic acid yield. In summary, an itaconic acid titer of 34 g/L with a total productivity of up to 0.07 g/L/h and a yield of 0.16 g/g could be reached during fed-batch cultivation. Conclusion: This study demonstrates the feasibility of consortium based CBP for itaconic acid production and also lays the fundament for the development and improvement of similar microbial consortia for cellulose based organic acid production.

Reduced by-product formation and modified oxygen availability improve itaconic acid production in Aspergillus niger

2013 ◽  
Vol 97 (9) ◽  
pp. 3901-3911 ◽  
Author(s):  
An Li ◽  
Nina Pfelzer ◽  
Robbert Zuijderwijk ◽  
Anja Brickwedde ◽  
Cora van Zeijl ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Itaconic Acid ◽  
Acid Production ◽  
Product Formation ◽  
Oxygen Availability

scholarly journals Integrated strain- and process design enable production of 220 g L−1 itaconic acid with Ustilago maydis

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Hamed Hosseinpour Tehrani ◽  
Johanna Becker ◽  
Isabel Bator ◽  
Katharina Saur ◽  
Svenja Meyer ◽  
...  
Keyword(s):  
Itaconic Acid ◽  
Acid Production ◽  
Process Development ◽  
Ustilago Maydis ◽  
Integrated Approach ◽  
Strain Engineering ◽  
Total Acid ◽  
Regulatory Cascade ◽  
Wide Range ◽  
Efficient Fermentation

Abstract Background Itaconic acid is an unsaturated, dicarboxylic acid which finds a wide range of applications in the polymer industry and as a building block for fuels, solvents and pharmaceuticals. Currently, Aspergillus terreus is used for industrial production, with titers above 100 g L−1 depending on the conditions. Besides A. terreus, Ustilago maydis is also a promising itaconic acid production host due to its yeast-like morphology. Recent strain engineering efforts significantly increased the yield, titer and rate of production. Results In this study, itaconate production by U. maydis was further increased by integrated strain- and process engineering. Next-generation itaconate hyper-producing strains were generated using CRISPR/Cas9 and FLP/FRT genome editing tools for gene deletion, promoter replacement, and overexpression of genes. The handling and morphology of this engineered strain were improved by deletion of fuz7, which is part of a regulatory cascade that governs morphology and pathogenicity. These strain modifications enabled the development of an efficient fermentation process with in situ product crystallization with CaCO3. This integrated approach resulted in a maximum itaconate titer of 220 g L−1, with a total acid titer of 248 g L−1, which is a significant improvement compared to best published itaconate titers reached with U. maydis and with A. terreus. Conclusion In this study, itaconic acid production could be enhanced significantly by morphological- and metabolic engineering in combination with process development, yielding the highest titer reported with any microorganism.

scholarly journals An Ustilago maydis chassis for itaconic acid production without by‐products

2019 ◽  
Vol 13 (2) ◽  
pp. 350-362 ◽  
Author(s):  
Johanna Becker ◽  
Hamed Hosseinpour Tehrani ◽  
Marc Gauert ◽  
Jörg Mampel ◽  
Lars M. Blank ◽  
...  
Keyword(s):  
Itaconic Acid ◽  
Acid Production ◽  
Ustilago Maydis ◽  
By Products

High level production of itaconic acid at low pH by Ustilago maydis with fed-batch fermentation

2021 ◽  
Author(s):  
Hatice Taşpınar Demir ◽  
Emine Bezirci ◽  
Johanna Becker ◽  
Hamed Hosseinpour Tehrani ◽  
Emrah Nikerel ◽  
...  
Keyword(s):  
Itaconic Acid ◽  
Batch Fermentation ◽  
Ustilago Maydis ◽  
Low Ph ◽  
Fed Batch ◽  
Fed Batch Fermentation ◽  
High Level ◽  
Level Production
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