Robust succinic acid production from crude glycerol using engineered Yarrowia lipolytica

Abstract

BackgroundIntegrating waste management with fuels and chemical production is considered to address the food waste problem and oil crisis. Approximately, 600 million tonnes crude glycerol is produced from the biodiesel industry annually, which is a top renewable feedstock for succinic acid production. To meet the increasing demand for succinic acid production, the development of more efficient and cost-effective production methods is urgently needed. Herein, we have proposed a new strategy for integration of both biodiesel and SA production in a biorefinery unit by construction of an aerobic yeast Yarrowia lipolytica with a deletion in the gene coding succinate dehydrogenase subunit 5.ResultsRobust succinic acid production by an engineered yeast Y. lipolytica from crude glycerol without pre-treatment was demonstrated. Diversion of metabolic flow from tricarboxylic acid cycle led to the success in generating a succinic acid producer Y. lipolytica PGC01003. The fermentation media and conditions were optimized, which resulted in 43 g L−1 succinic acid production from crude glycerol. Using the fed-batch strategy in 2.5 L fermenter, up to 160 g L−1 SA was yielded, indicating the great industrial potential.ConclusionsInactivation of SDH5 in Y. lipolytica Po1f led to succinic acid accumulation and secretion significantly. To our best knowledge, this is the highest titer obtained in fermentation on succinic acid production. In addition, the performance of batch and fed-batch fermentation showed high tolerance and yield on biodiesel by-product crude glycerol. All these results indicated that PGC01003 is a promising microbial factorial cell for the highly efficient strategy solving the environmental problem in connection with the production of value-added product.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0597-8) contains supplementary material, which is available to authorized users.

Characterization of threeYarrowia lipolyticastrains in respect to different cultivation temperatures and metabolite secretion

“…The first two media are commercially available and contain a balanced amount of carbon and nitrogen sources. They are broadly used for propagation of yeasts and with modified formulation for protein, lipid and organic acid production with Y. lipolytica (Madzak, Tréton and Blanchin-Roland 2000;Beopoulos et al 2008;Gao et al 2016) . In this work both media contained 20 g L -1 glucose.…”

A review of progress in (bio)catalytic routes from/to renewable succinic acid

“…More recently, engineered Yarrowia lipolytica strains (with gene-encoding succinate dehydrogenase subunit 5 being deactivated to block respiration pathway to fumarate) showed signifi cant improvements, with SA batch production reaching titers up to 43 g L -1 from 200 g L -1 crude glycerol media (realistic titers which allow integration with a commercial biodiesel plant). 40 When grown in 2.5 L fed-batch experiments, volumetric productivities of 0.4 g L -1 h -1 were achieved. Th e pH ranged between 4 and 6 and was adjusted with NaOH with aeration feed ca.…”

“…In comparison, fermentation with E. coli ATCC8739 of 10% crude glycerol (obtained from Sunn Hemp seed extraction and transesterification) yielded 1.4 g L −1 SA, while the purified glycerol feed gave 23.3 g L −1 and the commercial glycerol feed 30.8 g L −1 . More recently, engineered Yarrowia lipolytica strains (with gene‐encoding succinate dehydrogenase subunit 5 being deactivated to block respiration pathway to fumarate) showed significant improvements, with SA batch production reaching titers up to 43 g L −1 from 200 g L −1 crude glycerol media (realistic titers which allow integration with a commercial biodiesel plant) . When grown in 2.5 L fed‐batch experiments, volumetric productivities of 0.4 g L −1 h −1 were achieved.…”

Understanding Functional Roles of Native Pentose-Specific Transporters for Activating Dormant Pentose Metabolism inYarrowia lipolytica

“…It has an efficient and robust native metabolism to produce high levels of organic acids and neutral lipids. Combining metabolic engineering and fermentation optimization, engineered Y. lipolytica strains have been generated for high production of citric acid (titer: 101.0 g/L, rate: 0.42 g/L/h; yield: 0.89 g/g glucose) (5), alpha-ketoglutaric acid (186 g/L, 1.75 g/L/h, 0.36 g/g glycerol) (6), succinic acid (160.2 g/L, 0.4 g/L/h, 0.4 g/g crude glycerol) (7), and neutral lipids (84.5 g/L, 0.73 g/L/h, 0.20 g/g glucose) (8). In addition, Y. lipolytica exhibits exceptional robustness to tolerate in inhibitory environments associated with biomass pretreatments.…”