New source of cellulase production using a metagenomic technique
M. Ali, S., A. Soliman, N. (2019). New source of cellulase production using a metagenomic technique. EKB Journal Management System, 3(6), 590-597. doi: 10.21608/nrmj.2019.66751
Safaa M. Ali; Nadia A. Soliman. "New source of cellulase production using a metagenomic technique". EKB Journal Management System, 3, 6, 2019, 590-597. doi: 10.21608/nrmj.2019.66751
M. Ali, S., A. Soliman, N. (2019). 'New source of cellulase production using a metagenomic technique', EKB Journal Management System, 3(6), pp. 590-597. doi: 10.21608/nrmj.2019.66751
M. Ali, S., A. Soliman, N. New source of cellulase production using a metagenomic technique. EKB Journal Management System, 2019; 3(6): 590-597. doi: 10.21608/nrmj.2019.66751

New source of cellulase production using a metagenomic technique

Novel Research in Microbiology Journal
Volume 3, Issue 6, November and December 2019, Page 590-597  XML PDF (218.94 K)
Document Type: Original Article
DOI: 10.21608/nrmj.2019.66751
View on SCiNiTO View on SCiNiTO
Authors
Safaa M. Ali email orcid 1; Nadia A. Soliman2
1Department of Nucleic Acid Research, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria, Egypt
2Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria, Egypt
Abstract
The cellulase enzymes with high effectiveness under conditions agreeable to the industrial processes necessities are one of the keys for the successful development of chemical and drug synthesis. The soil metagenome is an affluent source for the discovery of new natural products. The objective of the current study was to identify the isolated functional gene(s) of the cellulase enzyme by using metagenomics. The plan of work composed of collection of different soil samples, isolation of total DNA, fragmentation, cloning, and expression of the isolated gene(s) in the suitable host microorganism. The total genomic DNA was extracted using a kit (QIAGEN), and then digested by different restriction enzymes BamHI. The digested fragments ranging from ~300-5000 bp were ligated, cloned into pUC19 vector, and then transformed into Escherichia coli DH5α. The resulting clones were screened as cellulase producers using a qualitative method. The positive clones which showed hydrolysis on the plate were screened once more in Luria-Bertani (LB) medium. The plasmids were isolated and then tested using universal primer (M13), to detect the fragment size and sequence for the Polymerase Chain Reaction (PCR) products. This study establishes an effortless and professional method for cloning of recent cellulase genes through ecological metagenomes. In the outlook, the metagenomic guide approachs may be functional to the elevated selection of novel cellulase from the environment.
Keywords
Cellulase; Metagenome; Qualitative estimation; Plasmid; Cloning
Statistics
Article View: 113
PDF Download: 99