Expression of the Endoglucanase Gene in Escherichia coli for the Second-generation Bioethanol Production


  • Orapin Jantasaeng Thailand Institute of Scientific and Technological Research
  • Puntipa Thaenkudrua Thailand Institute of Scientific and Technological Research
  • Joo Shun Tan Universiti Sains Malaysia
  • Pongsathon Phapugrangkul Thailand Institute of Scientific and Technological Research


Bioethanol, Biomass, Endoglucanase, Cellulose


The endoglucanase enzyme in Escherichia coli was expressed to improve enzymatic activities at low temperature and facilitate large-scale enzyme production. The endoglucanase gene (GenBank CAE51308.1) encoding endoglucanase enzyme (EC was selected from Clostridium thermocellum, a major component in the cellulosome complex and effective in cellulose hydrolysis. The mature endoglucanase gene was cloned into pET28a (+) vector and expressed in E. coli BL21 (DE3) pLysS. SDS-PAGE revealed the molecular weight of recombinant endoglucanase enzyme at around 72 kDa. Carboxymethylcellulose (CMC) was used as a substrate to determine endoglucanase enzyme activity. Under various combinations of pH, temperature and time results showed that the recombinant enzyme reached maximum activity (3.05 U/mL) at pH 10, 40 °C for 10 min. Further study of large-scale enzyme production together with exoglucanase and β-glucosidase enzymes is planned for biomass hydrolysis in second-generation bioethanol production.


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How to Cite

Jantasaeng, Orapin, Puntipa Thaenkudrua, Joo Shun Tan, and Pongsathon Phapugrangkul. 2019. “Expression of the Endoglucanase Gene in Escherichia Coli for the Second-Generation Bioethanol Production”. Food and Applied Bioscience Journal 7 (3):27-35.