Screening and Characterization of a Cellulase Gene from the Gut Microflora of Abalone Using Metagenomic Library

Duwoon Kim; Se-Na Kim; Keun Sik Baik; Seong Chan Park; Chae Hong Lim; Jong-Oh Kim; Tai-Sun Shin; Myung-Joo Oh; Chi Nam Seong

doi:10.1007/s12275-011-0205-3

Articles

Page Path: HOME > J. Microbiol > Volume 49(1); 2011 > Article

Research Support, Non-U.S. Gov't
Screening and Characterization of a Cellulase Gene from the Gut Microflora of Abalone Using Metagenomic Library: Duwoon Kim ¹, Se-Na Kim ^2,3, Keun Sik Baik ², Seong Chan Park ², Chae Hong Lim ², Jong-Oh Kim ³, Tai-Sun Shin ³, Myung-Joo Oh ³, Chi Nam Seong ²; Journal of Microbiology 2011;49(1):141-145.
DOI: https://doi.org/10.1007/s12275-011-0205-3
Published online: March 3, 2011

¹Department of Food Science and Technology and Functional Food Research Center, Chonnam National University, Gwangju 500-757, Republic of Korea, ²Department of Biology, Sunchon National University, Suncheon 540-742, Republic of Korea, ³Korea Basic Science Institute, Suncheon Center, Suncheon 540-742, Republic of Korea

Corresponding author: Chi Nam Seong , Tel: +82-61-750-3613,

Received: 3 June 2010 • Accepted: 5 October 2010

prev next

201 Views
0 Download
11 Crossref
15 Scopus

Full Article

Download PDF

Abstract

A metagenomic fosmid library was constructed using genomic DNA isolated from abalone intestine. Screening of a library of 3,840 clones revealed a 36 kb insert of a cellulase positive clone (pAM11E10). A shotgun clone library was constructed using the positive clone (pAM11E10) and further screening of 3,840 shotgun clones with an approximately 5 kb insert size using a Congo red overlay revealed only one cellulase positive clone (pAM11L9). The pAM11L9 consisted of a 5,293-bp DNA sequence and three open reading frames (ORFs). Among the three ORFs, cellulase activity was only shown in the recombinant protein (CelAM11) coded by ORF3, which showed 100% identity with outer membrane protein A from Vibrio alginolyticus 12G01, but no significant sequence homology to known cellulases. The expressed protein (CelAM11) has a molecular weight of approximately 37 kDa and the highest CMC hydrolysis activity was observed at pH 7.0 and 37°C. The carboxymethyl cellulase activity was determined by zymogram active staining and different degraded product profiles for CelAM11 were obtained when cellotetraose and cellopentaose were used as the substrates, while no substrate hydrolysis was observed on oligosaccharides such as cellobiose and cellotriose.

Keywords: abalone;cellulase;metagenomic library;gut microflora

Figure & Data

References

Citations

Citations to this article as recorded by

Comparative genomics reveal distinct potential of Tamlana sp. S12 for algal polysaccharide degradation
Hai-Feng Xia, Xiao-Yu Jia, Yan-Xia Zhou, Zong-Jun Du, Da-Shuai Mu, Guan-Jun Chen
Frontiers in Marine Science.2023;[Epub] CrossRef
A primer on metagenomics and next‐generation sequencing in fish gut microbiome research
Tina Kollannoor Johny, Rinu Madhu Puthusseri, Sarita Ganapathy Bhat
Aquaculture Research.2021; 52(10): 4574. CrossRef
Characterization of Bacillus Species with Keratinase and Cellulase Properties Isolated from Feather Dumping Soil and Cockroach Gut
Sonika Sharma, Rajesh Kumar Prasad, Soumya Chatterjee, Angkita Sharma, Mohan G. Vairale, Kamlesh Kumar Yadav
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences.2019; 89(3): 1079. CrossRef
Prospects of Metagenomic Cellulases for Converting Lignocellulosic Biomass into Bio-ethanol
Sangeeta Pandey
Journal of Pure and Applied Microbiology.2017; 11(2): 1079. CrossRef
Metagenomics for the development of new biocatalysts to advance lignocellulose saccharification for bioeconomic development
Salvatore Montella, Antonella Amore, Vincenza Faraco
Critical Reviews in Biotechnology.2016; 36(6): 998. CrossRef
Identification and Characterization of Carboxyl Esterases of Gill Chamber-Associated Microbiota in the Deep-Sea Shrimp Rimicaris exoculata by Using Functional Metagenomics
María Alcaide, Anatoli Tchigvintsev, Mónica Martínez-Martínez, Ana Popovic, Oleg N. Reva, Álvaro Lafraya, Rafael Bargiela, Taras Y. Nechitaylo, Ruth Matesanz, Marie-Anne Cambon-Bonavita, Mohamed Jebbar, Michail M. Yakimov, Alexei Savchenko, Olga V. Golysh
Applied and Environmental Microbiology.2015; 81(6): 2125. CrossRef
Purification and characterization of a carboxymethyl cellulase from Artemia salina
Hyun Woo Zin, Kwang-Hyun Park, Tae Jin Choi
Biochemical and Biophysical Research Communications.2014; 443(1): 194. CrossRef
Cloning and functional characterization of endo-β-1,4-glucanase gene from metagenomic library of vermicompost
Muhammad Yasir, Haji Khan, Syed Sikander Azam, Amar Telke, Seon Won Kim, Young Ryun Chung
Journal of Microbiology.2013; 51(3): 329. CrossRef
Processive and nonprocessive cellulases for biofuel production—lessons from bacterial genomes and structural analysis
David B. Wilson
Applied Microbiology and Biotechnology.2012; 93(2): 497. CrossRef
Gene Cloning of Endoglucanase Cel5A from Cellulose-Degrading Paenibacillus xylanilyticus KJ-03 and Purification and Characterization of the Recombinant Enzyme
In-Hye Park, Jie Chang, Yong-Seok Lee, Shu-Jun Fang, Yong-Lark Choi
The Protein Journal.2012; 31(3): 238. CrossRef
Screening and characterization of a cellulase with endocellulase and exocellulase activity from yak rumen metagenome
Lei Bao, Qiang Huang, Lei Chang, Jungang Zhou, Hong Lu
Journal of Molecular Catalysis B: Enzymatic.2011; 73(1-4): 104. CrossRef

Cite this Article

Cite this Article: export Copy Download Format; Close

Download Citation

Download a citation file in RIS format that can be imported by all major citation management software, including EndNote, ProCite, RefWorks, and Reference Manager.

Format:

RIS — For EndNote, ProCite, RefWorks, and most other reference management software
BibTeX — For JabRef, BibDesk, and other BibTeX-specific software

Include:

Citation for the content below
Citation and abstract for the content below

Screening and Characterization of a Cellulase Gene from the Gut Microflora of Abalone Using Metagenomic Library

J. Microbiol. 2011;49(1):141-145. Published online March 3, 2011

DOI: https://doi.org/10.1007/s12275-011-0205-3

Related articles