Short ozonation for effective removal and detoxification of fermentation inhibitors resulting from thermal pretreatment

Yan Rosen, Alona Maslennikov, Beny Trabelcy, Yoram Gerchman*, Hadas Mamane

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Common pretreatment methods for bioethanol production from lignocellulosic biomass are carried out under acidic conditions at high temperatures and result in formation of by-products, such as formic acid, levulinic acid, furfural and HMF (5-hydroxymethylfurfural). These by-products have an inhibitory effect on yeasts, reducing yeast growth and fermentation, and thus ethanol production. In this study, the inhibitory dose of these by-products was investigated along with the use of short ozonation as a means of their detoxification. When the effect of ozone was tested directly on yeasts (without inhibitors), they exhibited tolerance to ozone, suggesting the utilization of ozonation in simultaneous saccharification and fermentation. Fifteen minutes of ozonation removed over 90% of the inhibitory effect originating from 50 mM furfural, 50 mM formic acid, 25 mM HMF, and 100 mM levulinic acid. Thermal pretreatment and enzymatic hydrolysis of pine sawdust resulted in inhibition of microaerobic yeast growth. This inhibition was relieved after short ozonation; however, continuous ozonation has motivated further inhibition, suggesting the formation of additional inhibitory compounds. The results demonstrate the applicability of short ozonation as a practical approach for removing by-products’ inhibitory effects originating from thermal pretreatment of lignocellulosic waste.

Original languageEnglish
Pages (from-to)1407-1418
Number of pages12
JournalRenewable Energy
StatePublished - Apr 2022


FundersFunder number
Environmental Protection Technologies Ltd.
Israeli Ministry of Environmental Protection132-3-4
Ministry of Science and Technology, Israel
Ministry of Energy, Israel214-11-006


    • Fermentation inhibitors
    • Formic acid
    • Furfural
    • HMF (5-hydroxymethylfurfural)
    • Levulinic acid
    • Thermal pretreatment


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