Mutagenic effect of acridine orange on the production of EPS in Bacillus sp., its applications as anticoagulant and genetic diversity by ISSR and SCoT technique

Document Type : Original Article

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Botany Department (Genetics), Faculty of Agriculture, Al-Azhar Univ., Cairo, Egypt

Abstract

Many important substances are produced by some organisms, such as extracellular polysaccharides (EPS). The genus Bacillus is an important one in this field. As a result, in this study, a defined strain of Bacillus sp < /em>. BAB3450 (Gen Bank) was used to produce EPS with the goal of increasing productivity by introducing acridine orange mutations (AO). The results were obtained from three mutations that were more productive than the wild type. On the other hand, using primers Inter-Simple Sequence Repeat (ISSR) and Start Codon Targeted Polymorphism (SCoT), a comparison with the three mutants and wild type were made at the molecular level. Eleven ISSR primers were used. The results showed that ISSR-19, with 92.86% polymorphism, was the best primer. Using ten SCoT primers, the results showed that SCoT-2 was the best primer, with 90% polymorphism. For the ISSR primer and SCoT primers, the wild type and three mutants were divided into two major groups based on marker analysis with distance 60 to 75% and 55 to 65%, respectively. Cluster analyses were used to create a dendrogram of Bacillus strains and mutants, revealing high genetic variations between the wild type and mutants. Finally, extracellular polysaccharides (EPS) have a wide range of industrial applications in biomedicine, such as anticoagulant. The EPS from wild type and three mutants were tested for anticoagulant comparing with heparin. Mutant No.1 gave the best anticoagulant (15˃%), while the wild type, mutant No.2, mutant No.3 and mutant No.4 gave 10˃% comparing with heparin (80˃%). 

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