Abstract
This study was aimed to purified and characterized the bacteriocin produced from Lactobacillus acidophilus HT1, in order to use it in a skin pharmaceutical formula. The optimal conditions for bacteriocin production was investigated and results showed that modified nutrient broth was the best medium with glucose (30 gm/L) and yeast extract (7 gm/L) with peptone (7 gm/L) were the optimum carbon and nitrogen sources. In addition, 2% inoculum size, 37C◦ and pH 6.4 were the optimal conditions to obtain maximum bacteriocin of 640 AU/ml after 24 hrs. The bacteriocin was purified using 70% ammonium salt saturation and gel filtration with sephadex G50 that resulted 20 % yield and 2560 AU/ml of activity, then the partial purified bacteriocin was characterized and found the bacteriocin was protein in nature and kept its activity after 10 min at 20, 30 and 40◦C, however 50% of the activity was lost at 50C◦. Moreover, it showed stability at pH 6 and 7 for 30 min whereas; no activity was observed at pH 4 and 9. In addition, results showed that bacteriocin has a bactericidal effect rather than bacteriostatic. A cream formula contained the bacteriocin was prepared which already examined in vitro and in vivo. The effectiveness of the formula was confirmed using Klebsiella sp., Staphylococcus aureus and Pseudomonas aeruginosa as indicator strains. Results established that treatment at the onset time was more effective and the time of healing was decreased.
Highlights
Among the lactic acid bacteria, Lactobacilli are the most commonly intestinal microbiota of vertebrates, including humans and involve themselves in fermentation of various foods, thereby improving the food quality and the health [10]
This study was focused on producing a bacteriocin from local isolate of Lactobacillus acidophilus with an activity against some common skin pathogens in order to introduce it in a pharmaceutical skin formula
Modified nutrient medium contain peptone and yeast extract which were subjected to optimization strategy in order to choose one nitrogen source that support both growth and bacteriocin production by L. acidophilus HT1
Summary
Among the lactic acid bacteria, Lactobacilli (the species of the genus Lactobacillus) are the most commonly intestinal microbiota of vertebrates, including humans and involve themselves in fermentation of various foods, thereby improving the food quality and the health [10]. MATERIALS AND METHODS Microorganism A local isolate of Lactobacillus acidophilus HT1 was used throughout this work This isolate was obtained from dairy products and human mouth samples and already identified through some biochemical tests and cultural characteristics as described by Bergey’s manual and via molecular detection using 16S ribosomal RNA gene (data not shown). Through preliminary tests, this isolate showed an ability to produce an active bacteriocin against several bacteria and was chosen to be used in this study. °C for 24 hrs (Memmert-Germany incubator) the samples were taken from each flask for the determination of bacteriocin activity and bacterial biomass which was measured as the dry weight of cell material [2]
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