Immobilization of Clostridium perfringens type D in calcium alginate beads: toxin production mimics free cell culture

  • Hakimeh Rakhshandeh Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 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
  • 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
Keywords: Clostridium perfringens; Cell-immobilization techniques; Encapsulation; Calcium alginate beads; Toxin pro- duction

Abstract

Background and Objectives: Cell-immobilization is used to maintain microbial culture to produce metabolites in repeat- ed-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 CaCl 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 load- ing 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 encapsu- lation method. Thus, it seems possible to produce toxins from beads engulfing C. perfringens on larger scales via repeat- ed-batch or continuous fermentation processes.

Published
2022-08-14
Section
Articles