Functional screening for cellulolytic activity in a metagenomic fosmid library of microorganisms associated with coral

• Published in: Genetics and molecular research Vol. 15; no. 4
• Main Authors: Sousa, F M O, Moura, S R, Quinto, C A, Dias, J C T, Pirovani, C P, Rezende, R P
• Format: Journal Article
• Language: English
• Published: Brazil 05.10.2016
• Subjects: Animals; Anthozoa - microbiology; Carboxymethylcellulose Sodium - metabolism; Cellulase - metabolism; Cellulose - metabolism; Electrophoresis, Agar Gel; Gene Library; Metagenomics; Restriction Mapping; Siderastrea stellata; Substrate Specificity
• ISSN: 1676-5680; 1676-5680
• DOI: 10.4238/gmr.15048770

Cellulases are enzymes that degrade cellulosic materials. Cellulose is the most abundant renewable carbon resource on Earth, and cellulases are used in various industrial sectors. Although cellulases are obtained from a variety of sources, this is the first description of cellulolytic activity isolated from a coral metagenomic library. A metagenomic fosmid library of microorganisms associated with the coral Siderastrea stellata, comprising 3552 clones, was screened for cellulolytic activity; this allows access to non-cultivable microorganisms by exploiting the full biotechnological potential. Clones were grown on LB agar plates supplemented with 0.5% carboxymethylcellulose and cellulase positive clones revealed by staining with Congo red. Using this approach, six positive clones with cellulolytic activity were identified. The enzymatic index (EI) of the positive clones was calculated by the ratio between the hydrolysis zone diameter and colony diameter. All positive clones had an EI greater than 1.5. Digestion of the DNA isolated from the six positive clones, using the HindIII restriction endonuclease, revealed different restriction patterns in each clone, indicating that the DNA of each clone is different. There is a growing interest for new cellulolytic enzymes in various industry sectors. Here, we present the initial selection of potential clones for cellulose degradation that could be targets for future studies of enzymatic characterization.