High-throughput screening of high-yield colonies of Rhizopus oryzae for enhanced production of fumaric acid

Huang, Lei; Wei, Peilian; Zang, Ru; Xu, Zhinan; Chen, Peilin

doi:10.1007/s13213-010-0039-y

Cited by 34 publications

(12 citation statements)

References 18 publications

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“…Studies conducted by our research group indicate that the ethanol production rate decreases significantly over time compared with the FA production rate, resulting in an increase in the FA yield over time [15]. This phenomenon is also present in the data of other authors [3,12,18,19], but has not been reported on. Manipulating the production phase to increase the fermentation time could lead to significant gains in FA yield.…”

Section: Introductionsupporting

confidence: 58%

Improved continuous fumaric acid production with immobilised Rhizopus oryzae by implementation of a revised nitrogen control strategy

Naude

Nicol

2018

New Biotechnology

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A novel fermentation system was employed whereby the mycelial mat of Rhizopus oryzae was attached to a polypropylene tube. Batch operation was used for growth, while continuous operation was employed during the fumaric acid production phase. A clear decrease in respiration, fumaric acid (FA) and ethanol production was observed when zero nitrogen was fed in the production phase, with FA productivity decreasing from an initial 0.7 g L h to 0.3 g L h after 150 h. With the addition of 0.625 mg L h of urea FA productivity dropped to only 0.4 g L h after 150 h and 0.3 g L h after 400 h. Under these conditions it was observed that the ethanol production rate decreased 20 times faster compared with the FA production rate, therefore resulting in high FA yields towards the end of the fermentation (instantaneous 0.96 g g and average 0.81 g g after 400 h). Increasing the urea feed rate to 1.875 mg L h resulted in a clear increase in FA production and respiration rates. This condition also resulted in a 25% increase in biomass after 150 h, while the decline in the ethanol production rate was seven times lower than in the 0.625 mg L h urea fermentation, resulting in lower FA yields.

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Section: Introductionsupporting

confidence: 58%

Improved continuous fumaric acid production with immobilised Rhizopus oryzae by implementation of a revised nitrogen control strategy

Naude

Nicol

2018

New Biotechnology

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“…Developing high performance production strains is another use for directed evolution techniques, combining random mutagenesis using UV radiation with high-throughput screening techniques: a strain able to produce 160% more fumaric acid than the wild type strain has been chosen [41].…”

Section: Directed Evolutionmentioning

confidence: 99%

Fumaric Acid Production: A Biorefinery Perspective

et al. 2018

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The increasing scarcity of fossil raw materials, together with the need to develop new processes and technology based on renewable sources, and the need to dispose of an increasing amount of biomass-derived waste, have boosted the concept of biorefineries. Both 1G and 2G biorefineries are focused on the obtention of biofuels, chemicals, materials, food and feed from biomass, a renewable resource. Fumaric acid, and most compounds involved in the Kreb cycle, are considered key platform chemicals, not only for being acidulants and additives in the food industry, but also for their prospective use as monomers. This review is focused on the biotechnological processes based on fungi, mainly of the Rhizopus genus, whose main product is fumaric acid, on the process conditions, the bioreactors and modes of operation and on the purification of the acid once it is produced.

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“…The lack of a powerful and versatile genetic toolbox for Rhizopus species has restricted wider biotechnological exploitation of this fungus. In previous studies on strain improvement, researchers have altered strains by mutagenesis using ultraviolet radiation or nitrosoguanidine treatment [ 2 , 13 – 15 ]. The first attempt to modify the biosynthetic pathway of fumaric acid in R. oryzae by metabolic engineering was in 2012 [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

Metabolic engineering of the thermophilic filamentous fungus Myceliophthora thermophila to produce fumaric acid

Chen

et al. 2018

Biotechnol Biofuels

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BackgroundFumaric acid is widely used in food and pharmaceutical industries and is recognized as a versatile industrial chemical feedstock. Increasing concerns about energy and environmental problems have resulted in a focus on fumaric acid production by microbial fermentation via bioconversion of renewable feedstocks. Filamentous fungi are the predominant microorganisms used to produce organic acids, including fumaric acid, and most studies to date have focused on Rhizopus species. Thermophilic filamentous fungi have many advantages for the production of compounds by industrial fermentation. However, no previous studies have focused on fumaric acid production by thermophilic fungi.ResultsWe explored the feasibility of producing fumarate by metabolically engineering Myceliophthora thermophila using the CRISPR/Cas9 system. Screening of fumarases suggested that the fumarase from Candida krusei was the most suitable for efficient production of fumaric acid in M. thermophila. Introducing the C. krusei fumarase into M. thermophila increased the titer of fumaric acid by threefold. To further increase fumarate production, the intracellular fumarate digestion pathway was disrupted. After deletion of the two fumarate reductase and the mitochondrial fumarase genes of M. thermophila, the resulting strain exhibited a 2.33-fold increase in fumarate titer. Increasing the pool size of malate, the precursor of fumaric acid, significantly increased the final fumaric acid titer. Finally, disruption of the malate–aspartate shuttle increased the intracellular malate content by 2.16-fold and extracellular fumaric acid titer by 42%, compared with that of the parental strain. The strategic metabolic engineering of multiple genes resulted in a final strain that could produce up to 17 g/L fumaric acid from glucose in a fed-batch fermentation process.ConclusionsThis is the first metabolic engineering study on the production of fumaric acid by the thermophilic filamentous fungus M. thermophila. This cellulolytic fungal platform provides a promising method for the sustainable and efficient-cost production of fumaric acid from lignocellulose-derived carbon sources in the future.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1319-1) contains supplementary material, which is available to authorized users.

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High-throughput screening of high-yield colonies of Rhizopus oryzae for enhanced production of fumaric acid

Cited by 34 publications

References 18 publications

Improved continuous fumaric acid production with immobilised Rhizopus oryzae by implementation of a revised nitrogen control strategy

Improved continuous fumaric acid production with immobilised Rhizopus oryzae by implementation of a revised nitrogen control strategy

Fumaric Acid Production: A Biorefinery Perspective

Metabolic engineering of the thermophilic filamentous fungus Myceliophthora thermophila to produce fumaric acid

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