Pseudomonas stutzeri CM1, Novel Thermotolerant Cellulase- Producing Bacteria Isolated from Forest Soil

Surachai Rattanasuk, Apisit Songsaeng and Thanatip Sriwarom

Background and Objective: Cellulase is an important enzyme that useful for agricultural residue hydrolysis such as plant stover, molasse, rice straw. Thermotolerant cellulases are required to apply in textile, food, detergent, biofuels and pharmaceutical applications. This research aimed to isolate the thermotolerant cellulase-producing bacteria from forest soil and to determine cellulase activity from isolated bacteria. Materials and Methods: Soil samples were collected from the Roi Et Rajabhat University forest. One gram of soil sample was mixed with Luria-Bertani (LB) broth medium and incubated at 37°C with shaking at 150 rpm for 24 h. The cultured broth was streaked on LB agar plate and incubated at 37°C for 24 h. Cellulase-producing bacteria were isolated using Carboxymethylcellulose (CMC) agar plate. Four bacterial isolates which presented a clear zone on CMC agar plate after flooded with iodine solution, named CM1, CM2, CM3 and CM4. Cellulase activity of 4 isolated bacteria was determined against various pH (pH 4-8) and temperature (50-100°C). Results: The results indicated that CM1 isolate showed the highest cellulase activity at 0.074 unit mL^–1 at 80°C and pH5. All isolates were identified using 16S rRNA gene sequencing. The results indicated that CM1, CM3 and CM4 were identified as Pseudomonas stutzeri. while isolate CM2 was Bacillus subtilis. Conclusion: This is the first report presenting the thermotolerant cellulase produced by Pseudomonas stutzeri. The thermotolerant cellulase produced from Pseudomonas stutzeri in this study will be useful in many industrial processes using cellulase at high temperatures.

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