Abstract
Effect of physical mutagens on the ability of Pseudomonas aeruginosa H3 in alginate production was performed by subjection of the cell suspension of P. aeruginosa H3 to different doses of UV radiation (2, 4, 6, 8 and 10 J/m2) and LASER for (30, 60, 90 and 120 sec.), then cell suspension was spread on LB agar plates and incubated at 37 ˚C for 24hrs. After that, random selection of thirty colonies that arose from the cells subjection to the effect of different doses of UV radiation and laser were made (90% of the suspended cells were killed and were tested for their alginate production). Results showed that the mutagenesis by UV radiation caused an increase in the ability of P. aeruginosa H3 in alginate production. Over-producer mutant, H3R1, was obtained from this treatment. The productivity of the alginate from this mutant was 170mg/l in comparison with the productivity of wild type (70mg/l). On the other hand, results also showed that mutagenesis using LASER radiation caused an increase in the ability of P. aeruginosa H3 in alginate production. Over-producer mutant, H3R42, was obtained from this treatment characterized by its high ability in alginate production (130mg/l) in comparison with the productivity of wild type (70mg/l). Results of Fourier Transform-Infra Red (FT-IR) for alginate produced by the wild type of P. aeruginosa H3 and over-producer mutants H3R1 and H3R42 at a wide range of wave lengths absorbance (400-4000) cm-1 showed that there were no any structural differences in the chemical structure of alginate produced by them on the basis of infra-red light absorption by different functional groups of alginate
Highlights
Alginate is a linear unbranched polymer of D- Mannuronic acid and its C5 epimer Lguluronic acid, which are linked by -1, 4-glycosidic bonds
In order to improve the ability of P. aeruginosa H3 in alginate production, this isolate was subjected to physical mutagenesis by UV and LASER radiation respectively
Results indicated in figure (1) showed the killing effect of UV on bacterial cells, total viable count of P. aeruginosa H3 was decreased from 1.31 x 109 CFU/ml in the zero time to 0.35x109 CFU/ml after the first exposure to 2J/m2 of UV radiation
Summary
Alginate is a linear unbranched polymer of D- Mannuronic acid and its C5 epimer Lguluronic acid, which are linked by -1, 4-glycosidic bonds. Alginates are one of the exopolysaccharides which act as extracellular material that allows the formation of differentiated biofilms, with restrict diffusion of clinical antibiotics and protect embedded cell against human antibacterial defense mechanisms [3]. These polysaccharides are commercially important because of the gelling and colloidal properties. It has a variety of uses, as an ingredient of photographic emulsions, dental impression material and as additive in various food stuff [4]. Because of the importance of alginate as a perfect microbial polysaccharide for different uses and applications, this study was aimed to enhance the ability of highly alginate producer isolate P. aeruginosa H3 in alginate production by physical mutagenesis using UV and LASER radiation
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