This short article describes preparation and characterization of beads of alginate
October 6, 2017
This short article describes preparation and characterization of beads of alginate containing probiotic bacteria of Lactobacillus acidophilus DMSZ20079. beads. Keywords: Lactobacillus acidophilus, Alginate, Bead, CaCl2, Hardening time Intro In line with a global tendency towards naturally happening providers, probiotics have progressively got more attention. Probiotics are live microorganisms (bacteria or yeasts), which when ingested or locally applied in sufficient figures confer one LH 846 or more specified demonstrated health benefits for the sponsor.1 Their most important benefits are classified as maintenance of normal intestinal microflora,2 defense against enteropathogen infections,3 controlling serum cholesterol levels,4 increasing lactose intolerance,5 and possessing anticarcinogenic and antimutagenic activities.6 To obtain the potential benefits of probiotics they ought to safely transit through acidic and enzymatic conditions of gastric tract and colonize and grow within the epithelium of colon in right population.7 According to FAOs guideline, probiotics should present at their active site in a minimum count of 106-7 CFU/g or ml. 1 To reach such viability different strategies have been employed so far. In this regard encapsulation of probiotics in wide variety of polymers is the most frequently applied method that is cited in numerous studies.8 Alginate, a popular material to encapsulate probiotics, is a naturally happening biocompatible and biodegradable linear anionic polysaccharide. Preparation of alginate bead, with well retained bacteria in their matrix, can be very easily achieved by simple techniques like extrusion or emulsion methods. 9 In spite of the wide software of calcium alginate microcapsules in this area, there are not any common agreement about the conditions used and various protocol in this regard have been published so far.10 The objective of this work was to evaluate the effect of the most important parameters in the preparation of calcium alginate beads including ALG concentration, CaCl2 concentration as well as hardening time within the size, morphology, encapsulation efficiency (EE) and acid viabilities of Lactobacillus acidophilus. Materials and Methods Materials L. acidophilus DSMZ20079 was from DSMZ (Germany), pepsin, pancreatin, sodium alginate, MRS broth and MRS agar, sodium hydrogen phosphate, calcium chloride, sodium LH 846 hydroxide and hydrochloric acid from Merck (Germany). Methods Preparation of inoculum LH 846 L. acidophilus was cultured in MRS broth at 37C for 18 hours. Tradition was harvested by centrifugation at 700 RCF at 4C for 7 min and washed twice with saline and collected by centrifugation as above. The washed bacterial cells were resuspended in 7 ml saline and the cell count was identified using pour plate technique LH 846 in MRS agar in triplicate. The cell suspension divided in some equivalent parts and consequently was used to prepare different formulations. Preparation of beads The extrusion technique was used to prepare ALG beads.11 Sodium alginate solution sterilized at 121C for 15 min. The cooled ALG remedy (20 ml) were mixed with bacterial inoculum and softly stirred for 30 min to obtain a homogeneous suspension. The suspensions were extruded dropwise through a 27 gage nozzle into sterile hardening remedy (CaCl2). The beads were shaken LH 846 at 150 rpm, isolated by aseptic filtration (Whatman No.1), washed twice with sterile water, and kept in 0.1% w/v pepton remedy at 4C. The prepared formulations are demonstrated in Table 1. Table 1 Compositions of the analyzed formulation Size and morphological analysis The particle size of beads was assessed using optical microscopy (Dino-lite, Taiwan) by Scion image analyzer software. Data were collected from 60 beads in each sample and mean particle size was reported. Element Ratio= Major axis/Minor axis. Encapsulation Effectiveness (EE) To determine the encapsulation effectiveness, firstly prepared beads were mechanically disintegrated in phosphate buffer (pH=6.8), then the quantity of entrapped cells after adequate dilution were measured by pour plate method and counts were expressed while quantity of colony forming devices (CFU), and calculated while: EE=(Log 10N /Log 10N0) 100 Where N is the quantity of viable entrapped cells released Rabbit polyclonal to PHC2 from your beads, and N0 is the quantity of free cells added to the biopolymer mixture immediately before the production process..