K-PAM: A Klebsiella Serotype Predictor and Capsular Antigen Modeler and Optimization of purification protocol of E. coli Wzi, an outer membrane protein

Karpagam, Uma Sudhakar (2017) K-PAM: A Klebsiella Serotype Predictor and Capsular Antigen Modeler and Optimization of purification protocol of E. coli Wzi, an outer membrane protein. Masters thesis, Indian Insitute of Technology Hyderbad.

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Abstract

Klebsiella , a Gram - negative bacterium accounts for most of the nosocomial infections. Resistance to carbapenem class of antibiotics in Klebsiella further adds complexity to the issue as they are considered “antibiotics of last resort”. Cell surface associated glycoconjugates are one of the major virulence traits of Gram - negative bacteria of which c apsular polysaccharides (CPS/K antigen) and l ipopolysaccharides (LPS) are of prime importance in Klebsiella . Both antigens contain repeating units of sugars with high variability. Targeting such antigens would require adequate knowledge of the composition and three dimensional structures. Herein , we provide a repository , K - PAM ( A Klebsiella Serotype Predi ctor and Capsular Antigen Model er ) for modeled three dimensional structures of K, O antigens and genetic loci information of the proteins involved in the synthesis and export of K antigens. K - PAM features a serotype predictor tool (based on NCBI Blast programmes), that could be helpful in iden tifying serotype of new strains. K - PAM also provides options for generation of multimer of the K antigen which could be used as model systems for development of drugs against these multidrug resistant bacteria and the database of Klebsiella antigens has been made accessible through www.iith.ac.in/ K - PAM / . Wzi, an outer membrane protein has been reported to bind the capsular polysaccharide onto the outer membrane of Gram - negative bacteria . Targeting the interaction between the protein and the antigen could pave way for novel antibacterial strategies. In this context, Wzi is cloned and protocol is optimized for purification of the protein in denaturing and non - denaturing conditions .

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