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Sequential optimization of the fermentation factors with integrating seed culture adaptation for increased biorefinery of beet molasses to lactic acid

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Abstract

Microbial bioconversion of agro-industrial wastes to green chemicals has become an important objective in industrial biotechnology for lowering the product costs. Therefore, in this work, beet molasses was used as a potential raw material for lactic acid (LA) production by a newly isolated bacterium, Enterococcus faecium S6. The pretreatment of molasses with EDTA has exhibited the highest effectiveness for bioconversion to LA. The optimal medium components and culture conditions were established. Supplementation of molasses by yeast extract (YE) and ammonium chloride have achieved LA production of 11.90 g/L at a yield of 0.71 g/g-consumed sugars and a productivity of 0.33 g/L/h. Various approaches were used for the optimization of relevant factors affecting l-LA production in batch fermentations. Using classical “one-factor-at-a-time” approach, the optimal factors were as follows: sugar concentration, 40 g/L; pH, 7.0; 40 °C; and inoculum size, 10% (v/v) achieving LA at 29.7 g/L. The effects of these five fermentation parameters were further investigated by response surface methodology (RSM) to maximize LA production. The optimized fermentation conditions by RSM were as follows: sugar concentration, 65 g/L; YE, 0.625 g/L; temperature, 42 °C; pH, 6.75; and inoculum size, 9.5% (v/v) that achieved LA production of 41.5 g/L. Further improvements were achieved using repeated batch fermentation that has conducted for 16 runs. Using seed culture pre-adapted to the actual fermentation conditions in repeated batch fermentation led to enhanced LA production with a shorter process time compared to batch fermentation. These results achieved a maximum LA of 64.7 g/L with high yield (0.94 g/g of consumed sugars) and productivity (2.16 g/L/h). This study presented a cost-effective and long-term fermentative production of LA from beet molasses.

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The datasets used and/or analyzed during the current study are available from the corresponding authors on request.

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Acknowledgments

The authors are greatly thankful to Dr. Ehab F.S. Elbelely for his support and assistance with statistical analysis and great contribution in this work. We would like to thank the members of Fermentation Biotechnology and Applied Microbiology Centre, Al-Azhar University for their great support and contributions throughout this work

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Authors and Affiliations

  1. Botany and Microbiology Department, Faculty of Science (Boys), Al-Azhar University, Nasr City, Cairo, PN 11884, Egypt

    Hassan M. A. Alrefaey, Mohamed Ali Abdel-Rahman, Saad El-Din Hassan & Mohamed Salah Azab

  2. Agriculture Microbiology Department, Soil, Water and Environment Research Institute (SWERI), Sakha Agricultural Research Station, Agriculture Research Center (ARC), Giza, Egypt

    Mohamed Nour El-Din

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  1. Hassan M. A. Alrefaey
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Correspondence to Mohamed Ali Abdel-Rahman.

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Highlights

- Biorefinery of beet molasses to high lactic acid (LA) production by E. faecium S6 was established.

- EDTA, yeast extract, and ammonium chloride improved LA fermentation from molasses.

- OFAT optimization approach achieved 29.7 g/L of LA.

- RSM optimization approach enhanced LA production to 41.5 g/L.

- Integrated seed adaptation with increased molasses concentration in repeated batch fermentation was established.

- Sequential optimization of relevant factors achieved 64.7 g/L of LA.

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Alrefaey, H.M.A., Abdel-Rahman, M.A., Hassan, S.ED. et al. Sequential optimization of the fermentation factors with integrating seed culture adaptation for increased biorefinery of beet molasses to lactic acid. Biomass Conv. Bioref. 11, 1013–1028 (2021). https://doi.org/10.1007/s13399-020-00773-3

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