Extractive fermentation as a novel strategy for high cell mass production of hetero-fermentative probiotic strain Limosilactobacillus reuteri

Authors: SELVAMANI, SHANMUGAPRAK - University Technology Malaysia; RAMLI, SOLLEH - University Technology Malaysia; DAILIN, DANIEL - University Technology Malaysia; NATASYA, KHAIRUN - University Technology Malaysia; VARZAKAS, THEODOROS - University Of Peloponnese; ABOMOELAK, BASSAM - Orlando Health Arnold Palmer Hospital; SUKMAWATI, DALIA - Jakarta State University; NURJAYADI, MUKTININGSIH - Jakarta State University; Liu, Siqing; GUPTA, VIJAI - Sruc-Scotland'S Rural College; EL ENSHASY, HESHAM - University Technology Malaysia

Submitted to: Fermentation
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/3/2022
Publication Date: 10/10/2022
Citation: Selvamani, S., Ramli, S., Dailin, D.J., Natasya, K.H., Varzakas, T., Abomoelak, B., Sukmawati, D., Nurjayadi, M., Liu, S., Gupta, V.K., El Enshasy, H.A. 2022. Extractive fermentation as a novel strategy for high cell mass production of hetero-fermentative probiotic strain Limosilactobacillus reuteri. Fermentation. 8(10). Article 527. https://doi.org/10.3390/fermentation8100527.
DOI: https://doi.org/10.3390/fermentation8100527

Interpretive Summary: Probiotics are live beneficial bacteria that are ingested to improve intestinal health. Probiotics have proven success in both human and animal studies. The global probiotic market was valued at $58 billion in 2021 and expected to reach over $110 billion by 2030. The success of the probiotic industries relies on the mass production of cultures and viable cells in the final products. In this study, we describe methods of cultivation that simultaneously remove inhibitory products, such as lactic acid, and acetic acid, so the cells can grow at a rapid rate and reach high cell densities. These techniques could be used as a low-cost strategy to ensure the market growth of this industry, which benefits consumers and livestock producers. In addition, probiotics are grown on sugars obtained from agricultural products so this work also helps agricultural growers.

Technical Abstract: This study reports a novel technique to enhance high cell mass and viable cell counts of heterofermentative probiotic strain - Limosilactobacillus reuteri. Cultivation of L. reuteri was incorporated with weak base anion exchange resins to remove the accumulating lactic acid in the fermentation broth. Two anion exchange resins - Amberlite IRA 67 and IRA 96 were tested highest adsorption capacity towards lactic acid. Batch fermentation and fed-batch cultivation were further analyzed with IRA 67 resins as the application resulted in higher maximum viable cells. The in-situ application of anion exchange resins found to create shear stress and does not promote growth of L. reuteri. Thus, an external and integrated resin column system was proposed. The viable cell count of batch fermentation incorporated with the integrated resin column was improved by 73 times (3.89 x 1011 ± 0.07 CFU mL-1) compared to control batch fermentation (5.35 x 109 ± 0.32 CFU mL-1) without the addition of resins. The growth improvement was achieved due to the highest adsorption of lactic acid was recorded by the integrated IRA 67 resin system that coupled with the stirred tank bioreactor batch fermentation.