1. Clostridia Enrichment Guangfei Liu HMC 2012 Enrichments & Isolations Presentation

2. Clostridium (Kloster/ , Spindle) 梭 Shuttle

3. Endospores Tortora et al. 2008. Microbiology An Introduction, 10th edition. Clostridium is a genus of Gram-positive bacteria, belonging to the Firmicutes. They are obligate anaerobes capable of producing endospores

4. The Enrichment

5. 37 ° C

6. The Enrichment

7. Pectins Utilization Pectins are linear polysaccharides derived from galactouronic acid units, where some of the carboxylic acid groups are esterified with methanol. During fermentation, the ester groups are hydrolyzed, and methanol is released Galactouronic acid Pectins

8. Methanol was the major nongaseous end product produced by some of the species examined. For C. buwricum strain 4P1 on pectin (mM): 16, methanol; 11.4, butyrate; 7.5, acetate; 2.3, ethanol; 0.8, lactate; and 0.7, isopropanol. The methanol produced by the species examined here was not further consumed. In control experiments, these species did not grow on methanol as energy source.

9. Inoculation and Incubation 80 °C 37 ° C

10. + CO 2 Fermentation of glucose by Clostridia. Glucose is degraded to pyruvate via the Embden-Meyerhof pathway Lengeler et al. (2009) Biology of the Prokaryotes GYT Broth (g/L): 10.0 glucose; 5.0 tryptone; 10.0 yeast extract; 0.5 Na-thioglycolate; 20.0 CaCO 3.

11. Glucose Butyrate - + 2CO 2 + 2H 2 + H + ΔG°’ = -255 kJ/mol glucose (85 kJ/mol ATP) Glucose + 2H 2 O 2Acetate - + 2CO 2 + 4H 2 + 2H + ΔG°’ = -206 kJ/mol glucose (52 kJ/mol ATP) 3 ATP 4 ATP Glucose 0.7 Butyrate - +0.6 Acetate - + 2CO 2 + 2.6 H 2 + 1.3 H + ΔG°’ = -240 kJ/mol glucose (71 kJ/mol ATP) 3.3 ATP

14. Acid Production and Catalase Test Negative Positive Propionic-acid-bacteria Clostridia

15. Thank you!