Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 14 4 2022 08 14 503 509 Hakimeh Rakhshandeh Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; Student Research Committee, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran Mehrdad Shamsaddini Bafti Anaerobic Bacterial Vaccines Research and Production Department, Kerman Branch, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Kerman, Iran Behnaz Familsatarian Cellular and Molecular Research Center, Research Institute for Prevention of Non-Communicable Disease, Qazvin University of Medical Sciences, Qazvin, Iran Maryam Nooshadokht Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran; Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran Payam Khazaeli Pharmaceutics Research Center and Faculty of Pharmacy, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran Omid Raiesi Department of Parasitology, School of Allied Medical Sciences, Ilam University of Medical Sciences, Ilam, Iran Bagher Amirheidari Pharmaceutics Research Center and Faculty of Pharmacy, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran; Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran 2021 03 17 2022 06 30 Background and Objectives: Cell-immobilization is used to maintain microbial culture to produce metabolites in repeated-batch or continuous fermentations, thereby reducing the time and resources spent on delivering mass production of microbe. The technique also enables shortening of the detoxification phase and the amount of formaldehyde required due to low incidence of viable bacteria in the extract. Materials and Methods: A solution of sodium alginate containing Clostridium perfringens cells was dropped into stirring CaCl2 solution via a sterile syringe needle. Optimizations resulted in reasonably uniform beads containing C. perfringens. Beads were externally stabilized by poly L-lysine, followed by immersion in a solution of Na-alginate to coat them with a new layer of alginate forming an alginate-PLL-alginate cortex. Results: This study proved successful in immobilizing C. perfringens cells inside uniform alginate microspheres. Cell loading and cell propagation inside the beads were measured. The cell loaded beads were cultivable in liquid media producing 550 minimum lethal doses per milliliter (MLD/ml) in a 72 h. Conclusion: The research paved the way for further investigations to optimize and establish an efficient bacterial encapsulation method. Thus, it seems possible to produce toxins from beads engulfing C. perfringens on larger scales via repeated-batch or continuous fermentation processes. https://ijm.tums.ac.ir/index.php/ijm/article/view/3047 https://ijm.tums.ac.ir/index.php/ijm/article/download/3047/1484