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

• Published in: Iranian journal of microbiology Vol. 14; no. 4; pp. 503 - 509
• Main Authors: Rakhshandeh, Hakimeh, Shamsaddini Bafti, Mehrdad, Familsatarian, Behnaz, Nooshadokht, Maryam, Khazaeli, Payam, Raiesi, Omid, Amirheidari, Bagher
• Format: Journal Article
• Language: English
• Published: Iran Tehran University of Medical Sciences 01.08.2022
• Subjects: Calcium alginate beads; Cell-immobilization techniques; Clostridium perfringens; Encapsulation; Original; Toxin production; Encapsulation; Cell-immobilization techniques; Calcium alginate beads; Toxin production; Clostridium perfringens
• ISSN: 2008-3289; 2008-4447
• DOI: 10.18502/ijm.v14i4.10236

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. A solution of sodium alginate containing cells was dropped into stirring CaCl solution via a sterile syringe needle. Optimizations resulted in reasonably uniform beads containing 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. This study proved successful in immobilizing 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. 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 on larger scales via repeated-batch or continuous fermentation processes.