Mks chemotaxonomy

Molecular Diagnostics Laboratory Department of Microbiology and Biotechnology Dr.M.K. SATEESH

Definition ,[object Object],[object Object],[object Object],[object Object],[object Object],Whole cell analysis Cell parts Techniques development Physical development Gas chrom- atography Dr.M.K. SATEESH

Key Terms ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Classification ,[object Object],[object Object],[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Chemical analysis Sophisticated tools are available for studying the structural composition of bacteria (most commonly fatty acid , carbohydrate or ubiquinone profiling) of bacteria. Characterization of secreted metabolic products (e.g. volatile alcohols and short chain fatty acids ) is also helpful. Dr.M.K. SATEESH

Major Chemical compounds ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Dr.M.K. SATEESH

A model for bacterial speciation Dr.M.K. SATEESH

[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Lipid composition ,[object Object],R− CO − O−Glycerol R−CO−O−CH 3 Methanolic Hcl Mycolic acids, polar lipids, phospholipids, glycolipids, hopanoids, hydrocarbons, carotenoids Dr.M.K. SATEESH

Examples ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],phospholipids Steroids Dr.M.K. SATEESH

Fatty acid methyl ester (FAME) analysis Dr.M.K. SATEESH

Isoprenoid Quiniones ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Cytochrome composition ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Cytochromes are membrane-bound hemoproteins that contain heme groups and carry out electron transport/redox processes-4 major types : a, b, c, d, o. Dr.M.K. SATEESH

Amino acids of proteins ,[object Object],[object Object],Cytochrome oxidase Dr.M.K. SATEESH

Cytochrome oxidase Comparison of cytochrome oxidase from bacteria and bovine Dr.M.K. SATEESH

Sequence comparison of cytochrome oxidase from three species Dr.M.K. SATEESH

Protein profiles/fingerprints Total Cellular membrane proteins PAGE/ Isoelectric focusing ~ 10-30 proteins ( water soluble) Clostridium, Rhizobium, Haemophilus Dr.M.K. SATEESH

Enzyme characterization Citrate synthase is found in all living cells, so it has been a useful enzyme for comparing differences from organism to organism. Citrate synthase Dr.M.K. SATEESH

Citrate synthase.(monomer shown ) citrate is shown in green, and CoA is pink. Dr.M.K. SATEESH

Succinyl-CoA synthetase, ( succinate thiokinase) succinyl-CoA synthetase, sometimes called succinate thiokinase Dr.M.K. SATEESH

Fermentation products profile ,[object Object],[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Oxidation - Reduction Reactions are Always Coupled Oxidation = A B H + H { { { Reduction = gain of e - & H + loss of e - & H + NAD + NADH + H = electron Dr.M.K. SATEESH B A

MKS Use of Intact Cell MALDI Fingerprints to Identify Bacillus and Pseudomonas Isolates

Introduction ,[object Object],[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Principals of MALDI ToF Mass Spectrometry ,[object Object],[object Object],Dr.M.K. SATEESH

Principles of MALDI ToF Mass Spectrometry ,[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Principles of MALDI ToF Mass Spectrometry: Bacterial Analysis ,[object Object],[object Object],[object Object],Dr.M.K. SATEESH

Mks chemotaxonomy

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