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Ergosterol as a primary metabolite and precursor of vitamin D2, is the most plentiful mycosterols in fungal cell membrane. Process optimization to increase the yield and productivity of biological products is a topic of interest. Ultrasonic waves have many applications in biotechnology, like cell disruption, and enhancement of primary and secondary metabolites production. This study disclosed an optimal condition for ultrasound-assisted production (UAP) of ergosterol from Penicillium brevicompactum MUCL 19,011 using L9 Taguchi statistical method. The intensity (IS), time of sonication (TS), treatment frequency (TF), and number of days of treatment (DT) were allocated to study the effects of ultrasound on ergosterol production. The results were analyzed using Minitab version 19. The maximum ergosterol, 11 mg/g cell dry weight (CDW), was produced on the tenth day while all factors were at a low level. The days of treatment with a contribution of 45.48% was the most significant factor for ergosterol production. For the first time, this study revealed the positive effect of ultrasound on the production of ergosterol. Ergosterol production increased 73% (4.63 mg/g CDW) after process optimization. Finally, a mathematical model of ultrasound factors with a regression coefficient of R2?=?0.978 was obtained for the ergosterol production during ultrasound treatment.

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