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Bacillus Bacterial Characteristics and Enzyme Hydrolysis Study

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Explore the enzymatic capabilities of Bacillus species such as amylase, lipase, casease, and gelatinase, along with hydrolysis reactions on starch, lipids, proteins, and collagen. Utilize Tri-Plate and unique agar tests for differentiation.

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Bacillus Bacterial Characteristics and Enzyme Hydrolysis Study

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  1. Bacillus

  2. Bacillus sp. • GP rod • Spore former • Examples • B. anthracis • B. cereus • B. subtilis • B. megatarium • Exoenzymes • Lipase • Amylase • Casease • Gelatinase

  3. Spores

  4. Agar Characteristics

  5. Starch Hydrolysis • Enzymes • Amylase • Maltase • Substrate • Starch • Reaction • Clear zone

  6. Protein Hydrolysis • Enzymes • Proteases • Caseases • Substance • Casein • Reaction • Clear zone

  7. Lipid Hydrolysis • Enzymes • Lipase • Substrate • Tributyrin • Reaction • Clear zone

  8. Bacillus Tri-Plate

  9. Collagen Hydrolysis • Enzyme • Gelatinase • Use loop (vs. needle) • Inoculate several times • Stab deep • Substrate • Gelatin • Reaction • Clear Zone on Agar • Liquefy Gelatin in tube

  10. MSA • Selective • Differential • Substrate • Mannitol • Salt (7.5%) • pH = phenol red • Reaction • Growth • Fermentation

  11. Nitrate Reduction • Inoculate with loop • Incubate • Additional solutions • Sulfanilic acid (A) • Naphthylamine (B) • (+) = Red • Nitrate to Nitrite • No color, add Zinc • (+) = not red • (-) = red

  12. Citrate • Enzyme • Citrase • Substrate • Citrate • pH • Bromthymol blue • Reaction • Keto acids • Oxaloacetic • Pyruvate • Acetic acid • CO2 • CO2 + NaH2CO3 • Prussian blue = +

  13. Bile Esculin Agar (BEA) • Selective • Bile Salts • Differential • Esculin (CH2O) • Fe-NH3-citrate • Reaction • Growth in bile • Hydrolyze esculin • Esculetin-Iron Rxn • Black precipitate

  14. Results • Exoenzymes • Starch (amylase, maltase) • Lipid (lipase) • Proteins (Casease, protease) • Collagen (Gelatinase) • Reaction • (+) = clearing • (-) = no clearing

  15. Starch Hydrolysis

  16. Lipid Hydrolysis

  17. Protein Hydrolysis

  18. Gelatin (Collagen) Hydrolysis

  19. BEA and Citrate Results + + Bile Esculin Citrate

  20. Hydrolysis Results Summary casein lipid Gelatin Starch

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