Cloning and expression of Ruminococcus flavefaciens cellulase-encoding genes in Escherichia coli Khairil Anwar (a), Sukarne (a), Lalu Unsunnidhal (b), Muhammad Aidil Fitriyan Fadjar Suryadi (a), Zaid Al Gifari (c), Alimuddin (a,d), Muhamad Amin (e), Made Sriasih (a), Muhamad Ali (a)*
a)Laboratory of Biotechnology and Animal Product Processing, Faculty of Animal Sciences, University of Mataram, Mataram, Indonesia, *m_ali[at]unram.ac.id b)Faculty of Food Science and Technology, University of Mataram, Mataram, Indonesia, c)Laboratory of Animal Production, Faculty of Animal Sciences, University of Mataram, Mataram, Indonesia, d)Faculty of NW Mataram, Mataram, Indonesia, e) Fish Nutrition Research Group, Department of Aquaculture, Faculty of Fisheries and Marine, Universitas Airlangga, Surabaya, Indonesia
Abstract
Carbohydrate-metabolizing enzymes are crucial in improving feed digestibility and efficiency in the poultry industry. Among these, cellulase, which breaks down cellulose, is particularly valued across various industries, especially in animal feed. However, producing cellulase in its native host, such as Ruminococcus flavefaciens, is often challenging due to complex cultivation requirements and limited yields. In this study, the cellulose-encoding gene from Ruminococcus flavefaciens was synthesized and expressed in Escherichia coli. The gene was optimized and cloned into pET15b vector suitable for the E. coli expression system. The results showed a protein band at approximately 38 kDa, matching the expected size of the cellulase enzyme produced in E. coli. The optimized codon usage for E. coli significantly enhanced the soluble fraction of cellulase, which is crucial for large-scale production. In summary, the recombinant Ruminococcus flavefaciens cellulose-encoding gene was successfully expressed in E. coli. Although the expression of cellulase in E. coli was successful, further optimization is needed to increase yields and assess enzymatic activity.
Keywords: carbohydrate-metabolizing enzyme, poultry industry, Ruminococcus flavefaciens cellulase, recombinant protein, codon optimization, E. coli expression system
