ISOLATION AND DETERMINATION OF CHROMIUM (VI) AND NICKEL (II) RESISTANT SIDEROPHORE PRODUCING BACILLUS CEREUS(sp) AND ANALYSIS OF THE PROTEIN EXPRESSION USING BIOINFORMATICS TOOLS

Abstract

Siderophores are produced by bacteria, fungi and plants to facilitate uptake of iron, they are low-molecular-weight chelation agents and protect from stress responses. Pyoverdines act as fluorescent siderophores, plays an important role under iron-limiting conditions. During the stress condition a change in the broth culture was observed where it turned green from yellow, indicating some kind of pigment release. According to some reports the pigment release happens due to the presence of Siderophore in microorganisms. This colour change can also be considered as microbial sensor for color change detection. The bacterial sample was tested for the chromium (VI) and Nickel (II) resistant capacity, presence of siderophore, DNA and protein content. The chromium (VI) and nickel (II) resistant isolates were identified as Bacillus cereus (BF2). The siderophore was detected using qualitative and quantitative assays. Using various bioinformatics softwares the siderophore produced by Bacillus cereus (Rock 3-28) was compared with the isolated culture because both of their molecular weight varied from the range of 48kDa-65kDa. National Center For Biotechnology Information (NCBI), Basic Local Alignment Search Tool (BLAST), Clustal Omega, Expert Protein Analysis System (ExPASy), Self-Optimized Prediction Method with Alignment (SOPMA) and SWISS-MODEL tools were used to study the pyoverdine protein which coded for the enzyme L-ornithine N5-oxygenase. The FASTA sequences of the query sequence were compared with other species using BLAST. The sequences of protein were used for structural and functional analysis and for phylogenetic tree construction.