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Towards closed carbon loop fermentations: Cofeeding of Yarrowia lipolytica with glucose and formic acid.

ABSTRACT: A novel fermentation process was developed in which renewable electricity is indirectly used as an energy source in fermentation, synergistically decreasing both the consumption of sugar as a first generation carbon source and emission of the greenhouse gas CO₂ . As an illustration, a glucose-based process is co-fed with formic acid, which can be generated by capturing CO₂ from fermentation offgas followed by electrochemical reduction with renewable electricity. This "closed carbon loop" concept is demonstrated by a case study in which cofeeding formic acid is shown to significantly increase the yield of biomass on glucose of the industrially relevant yeast species Yarrowia lipolytica. First, the optimal feed ratio of formic acid to glucose is established using chemostat cultivations. Subsequently, guided by a dynamic fermentation process model, a fed-batch protocol is developed and demonstrated on laboratory scale. Finally, the developed fed-batch process is tested and proven to be scalable at pilot scale. Extensions of the concept are discussed to apply the concept to anaerobic fermentations, and to recycle the O₂ that is co-generated with the formic acid to aerobic fermentation processes for intensification purposes.

SUBMITTER: van Winden WA

PROVIDER: S-EPMC9540030 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature

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Towards closed carbon loop fermentations: Cofeeding of Yarrowia lipolytica with glucose and formic acid.

van Winden Wouter A WA Mans Robert R Breestraat Stefaan S Verlinden Rob A J RAJ Mielgo-Gómez Álvaro Á de Hulster Erik A F EAF de Bruijn Hans M C J HMCJ Noorman Henk J HJ

Biotechnology and bioengineering 20220507 8

A novel fermentation process was developed in which renewable electricity is indirectly used as an energy source in fermentation, synergistically decreasing both the consumption of sugar as a first generation carbon source and emission of the greenhouse gas CO<sub>2</sub> . As an illustration, a glucose-based process is co-fed with formic acid, which can be generated by capturing CO<sub>2</sub> from fermentation offgas followed by electrochemical reduction with renewable electricity. This "close ...[more]

PMID: 35451059

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Project description:BACKGROUND:Yarrowia lipolytica is an oleaginous yeast capable of metabolizing glucose to lipids, which then accumulate intracellularly. However, it lacks the suite of cellulolytic enzymes required to break down biomass cellulose and cannot therefore utilize biomass directly as a carbon source. Toward the development of a direct microbial conversion platform for the production of hydrocarbon fuels from cellulosic biomass, the potential for Y. lipolytica to function as a consolidated bioprocessing strain was investigated by first conducting a genomic search and functional testing of its endogenous glycoside hydrolases. Once the range of endogenous enzymes was determined, the critical cellulases from Trichoderma reesei were cloned into Yarrowia. RESULTS:Initially, work to express T. reesei endoglucanase II (EGII) and cellobiohydrolase (CBH) II in Y. lipolytica resulted in the successful secretion of active enzymes. However, a critical cellulase, T. reesei CBHI, while successfully expressed in and secreted from Yarrowia, showed less than expected enzymatic activity, suggesting an incompatibility (probably at the post-translational level) for its expression in Yarrowia. This result prompted us to evaluate alternative or modified CBHI enzymes. Our subsequent expression of a T. reesei-Talaromyces emersonii (Tr-Te) chimeric CBHI, Chaetomium thermophilum CBHI, and Humicola grisea CBHI demonstrated remarkably improved enzymatic activities. Specifically, the purified chimeric Tr-Te CBHI showed a specific activity on Avicel that is comparable to that of the native T. reesei CBHI. Furthermore, the chimeric Tr-Te CBHI also showed significant synergism with EGII and CBHII in degrading cellulosic substrates, using either mixed supernatants or co-cultures of the corresponding Y. lipolytica transformants. The consortia system approach also allows rational volume mixing of the transformant cultures in accordance with the optimal ratio of cellulases required for efficient degradation of cellulosic substrates. CONCLUSIONS:Taken together, this work demonstrates the first case of successful expression of a chimeric CBHI with essentially full native activity in Y. lipolytica, and supports the notion that Y. lipolytica strains can be genetically engineered, ultimately by heterologous expression of fungal cellulases and other enzymes, to directly convert lignocellulosic substrates to biofuels.

Dataset Information

Towards closed carbon loop fermentations: Cofeeding of Yarrowia lipolytica with glucose and formic acid.

Publications

Towards closed carbon loop fermentations: Cofeeding of Yarrowia lipolytica with glucose and formic acid.

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