A Novel: Low-cost Method for the Isolation of Anaerobic Hydrogen Producing Bacteria

Abstract

Abstract The hydrogen content of the two sludge samples namely TDS and PLS were increased after the heat shock pretreatment from 239.23 ± 10.23 mL/L to 663.87 ± 15.99 mL/L at 80°C and from 173.61 ± 17.21 mL/L to 496.74 ± 30.21 mL/L at 60°C, respectively. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) indicated that Enterococcus and Clostridium were the dominant bacteria in the TDS and PLS consortium. Two strains were successfully isolated from TDS and PLS using a simple method. Based on Bergey’s key identification and 16S rDNA sequencing, they were identified as Clostridium butyricum strain TD and Clostridium acetobutylicum strain PL. The hydrogen content, glucose consumed yield, and hydrogen yield of C. butyricum TD and C. acetobutylicum PL were 545.24 ± 7.31 mL/L, 90 ± 0.41 %, 0.89 ± 0.01 mol H2/mol glucose, and 729.34 ± 22.81 mL/L, 92.77 ± 1.39 %, 1.26 ± 0.05 mol H2/mol glucose, respectively.