227703

New source of cellulase production using a metagenomic technique

Article

Last updated: 05 Jan 2025

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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.

DOI

10.21608/nrmj.2019.66751

Keywords

Cellulase, Metagenome, Qualitative estimation, plasmid, cloning

Authors

First Name

Safaa

Last Name

M. Ali

MiddleName

-

Affiliation

Department of Nucleic Acid Research, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria, Egypt

Email

safaa.mohamedali@yahoo.com

City

-

Orcid

0000-0001-9850-0174

First Name

Nadia

Last Name

A. Soliman

MiddleName

-

Affiliation

Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria, Egypt

Email

-

City

-

Orcid

-

Volume

3

Article Issue

6

Related Issue

32627

Issue Date

2019-12-01

Receive Date

2019-12-04

Publish Date

2019-12-01

Page Start

590

Page End

597

Print ISSN

2537-0286

Online ISSN

2537-0294

Link

https://nrmj.journals.ekb.eg/article_227703.html

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https://nrmj.journals.ekb.eg/service?article_code=227703

Order

227,703

Type

Original Article

Type Code

2,265

Publication Type

Journal

Publication Title

Novel Research in Microbiology Journal

Publication Link

https://nrmj.journals.ekb.eg/

MainTitle

New source of cellulase production using a metagenomic technique

Details

Type

Article

Created At

23 Jan 2023