Construction and Cloning of the CelE Gene Encoding the Cellulase Enzyme as A Candidate Enzyme For the Food and Agroindustry
DOI:
https://doi.org/10.29303/profood.v10i1.370Keywords:
CelE, Cellulase Enzyme, Recombinant DNA Cloning, Recombinant DNA ConstructionAbstract
Research on the construction and cloning of the CelE gene, which encodes the cellulase enzyme from Ruminococcus plavefaciens, emerged as a response to the urgent need for efficient enzyme resources in the food industry and agro-industry. Cellulase enzymes, with a focus on CelE, have a vital role in the degradation process of cellulose, a main component in plant cell walls. The existence of Ruminococcus plavefaciens as a source of the CelE gene is of interest because this microorganism is found in the digestive system of ruminant animals and has great potential to produce efficient cellulase enzymes. This research aimed to create and clone the pET15b plasmid with the CelE gene. Confirmation of the CelE gene in recombinant DNA was carried out by identifying host bacterial resistance in media with antibiotics and colony PCR. This research was carried out by applying the following methods: Implementation of Codon Optimization and Recombinant Plasmid Construction (CelE), Preparation of Competent Cells, Cell Transformation, and Transformant Colony PCR Test. The results of the research show that this research has succeeded in obtaining the expected transformant bacteria with the results of the Transformant Colony PCR Test. obtained the appropriate product size, namely 205 bp, so it can be concluded that the pET15b plasmid with the optimized CelE gene was successfully constructed and cloned.
References
Arthur, P.K., Alvarado, L.J. & Dayie, T.K. (2011) Expression, purification and analysis of the activity of enzymes from the pentose phosphate pathway. Protein Expression and Purification. 76 (2), 229–237. doi:10.1016/j.pep.2010.11.008.
Barokah, U., Unsunnidhal, L., Kristianingrum, Y.P. & Kusumawati, A. (2023) Transformation of HBcAg gene of hepatitis B virus using pEGFP-C1 vector. In: THE 5TH INTERNATIONAL CONFERENCE ON BIOSCIENCE AND BIOTECHNOLOGY. 2023 Mataram, AIP Publishing. p. doi:https://doi.org/10.1063/12.0020985.
Datta, R. (2024) Enzymatic degradation of cellulose in soil: A review. Heliyon. 10 (1), e24022. doi:10.1016/j.heliyon.2024.e24022.
Fathurrahman, I., Kusumawati, A., Rahman, A., Ulviani, Y., Prihantoko, K.D. & Unsunnidhal, L. (2022) Molecular sexing in Bos taurus using quantitative polymerase chain reaction (qPCR) method. IOP Conference Series: Earth and Environmental Science. 976 (1). doi:10.1088/1755-1315/976/1/012002.
Hua, D., Hendriks, W.H., Xiong, B. & Pellikaan, W.F. (2022) Starch and Cellulose Degradation in the Rumen and Applications of Metagenomics on Ruminal Microorganisms. Animals. 12 (21), 1–13. doi:10.3390/ani12213020.
Ishak, J., Unsunnidhal, L., Martien, R. & Kusumawati, A. (2019) In vitro evaluation of chitosan-DNA plasmid complex encoding Jembrana disease virus Env-TM protein as a vaccine candidate. Journal of Veterinary Research. 63 (1), 7–16. doi:https://doi.org/10.2478/jvetres-2019-0018.
Jannah, R. & Unsunnidhal, L. (2019) KONSTRUKSI DAN KLONING PLASMID PCDNA3 . 1 ( + ) DENGAN SUBGENOTIP B3 HEPATITIS B CORE ANTIGEN ( HBcAg ) SEBAGAI KANDIDAT VAKSIN DNA HEPATITIS B. Jurnal Penelitian dan Kajian Ilmiah Kesehatan. 5 (2), 125–131.
Rahman, A., Kusumawati, A., Budiyanto, A., Ulviani, Y., Fathurrahman, I., Prihantoko, K.D. & Unsunnidhal, L. (2022) Molecular Verification of Sex-separated Straw of Simmental Cattle ( Bos taurus ) by Polymerase Chain Reaction (PCR) . Proceedings of the 9th International Seminar on Tropical Animal Production (ISTAP 2021). 18 (Istap 2021), 223–226. doi:10.2991/absr.k.220207.046.
Tuttle, A.R., Trahan, N.D. & Son, M.S. (2021) Growth and Maintenance of Escherichia coli Laboratory Strains. Current Protocols. 1 (1), 1–13. doi:10.1002/cpz1.20.
Unsunnidhal, L., Ishak, J. & Kusumawati, A. (2019) Expression of gag-CA Gene of Jembrana Disease Virus with Cationic Liposomes and Chitosan Nanoparticle Delivery Systems as DNA Vaccine Candidates. Tropical Life Sciences Research. 30 (3), 15–36. doi:https://doi.org/10.21315/tlsr2019.30.3.2.
Unsunnidhal, L. & Jannah, R. (2019) POTENTIAL OF CATIONIC LIPOSOMES AND CHITOSAN NANOPARTICLES FOR DELIVERY DNA VACCINE MODEL NTC8685-EGFP. Jurnal Penelitian dan Kajian Ilmiah Kesehatan. 5 (2), 120–124.
Unsunnidhal, L., Jannah, R., Haris, A., Supinganto, A. & Kusumawati, A. (2021a) Potential of Nanoparticles Chitosan for Delivery pcDNA3.1-SB3- HBcAg. BIO Web of Conferences. 41 (07003), 1–6.
Unsunnidhal, L., Perdhana, F.F., Utama, Q.D. & Jannah, R. (2023) Identifikasi Kandungan Daging Sapi Menggunakan Polymerase Chain Reaction (PCR) Pada Pedagang Bakso Tusuk Keliling Di Lingkungan Kota Mataram, Nusa Tenggara Barat. Pro Food. 9 (1), 76–81. doi:10.29303/profood.v9i1.312.
Unsunnidhal, L., Wasito, R., Nugraha Setyawan, E.M., Warsani, Z. & Kusumawati, A. (2021b) Potential of polylactic-co-glycolic acid (PLGA) for delivery Jembrana disease DNA vaccine Model (pEGFP-C1-tat). Journal of Veterinary Science. 22 (6), 1–15. doi:10.4142/jvs.2021.22.e76.
Unsunnidhal, L., Wasito, R., Setyawan, E.M.N. & Kusumawati, A. (2021c) Potential of Nanoparticles Chitosan for Delivery pcDNA3.1-tat. BIO Web of Conferences. 41 (07004), 1–6.
