The isolation and identification of hydrogen production bacteria(HPB)with high yield and high evolution rate is an important foundation of fermented hydrogen production process by anaerobic digesting high strength organic wastewater. An improved Hungater rolling tubes technique and a plate method of culture bottle (PMCB) were employed to enumerate and isolate the anaerobes. The HPB-RL medium was designed specially for isolating and culturing anaerobic hydrogen bacteria. Culture temperature was at 37°C and 5.5 of pH. About 90 strains of bacteria were isolated, in which, a strain of the hydrogen producer with high yield and high evolution rate was isolated and named Rennanqi12. The isolates were straight rods, Grams positive, strictly anaerobic, no spore-forming bacteria, whole bacteria flagella, having capsule, 3~4 metachromatic granules. The strain of Rennanqi12 could produce hydrogen using molasses wastewater as the carbon resource. The isolate Rennanqi12 has the closest relationship with the strain C. cellulose for the 94% homology of 16S rDNA. The strain of Rennanqi12 might be a new species that belongs to a new genus. The amount of hydrogen-producing (YH2)is 1880.5ml/L medium and the maximum hydrogen-producing rate(QH2)is 28.2 mmol/g•(dry•cell)•h when R12 digested substrates were glucose.