Cloning of genes encoding for cellulolytic enzymes for metagenome in goat's rumen

Santosh Thapa, Tennessee State University


The paucity of enzymes that efficiently deconstruct plant polysaccharides represents a major bottleneck for industrial-scale conversion of cellulosic biomass into biofuels. The cellulosic bioconversion into fermentable sugar is a complex process and requires the synergistic action of endoglucanases, exoglucanases and ?-glucosidases. These enzymes are secreted by the ruminal microbes in ruminant animals such as goats. To be able to tap into those resources is expected to have a significant impact on cellulosic biofuel production, but a majority of those microbial species are not culturable under laboratory conditions (using laboratory culture techniques). This study was performed to develop molecular approaches for using these genome resources. Metagenome DNA was extracted and sequenced using next-generation sequencing. Following assembly and annotation, 14 gene clones encoding for cellulases, hemicellulases and xylanases have been cloned from the metagenome. PCR primers were designed against the assembled gene sequences, and PCR reactions were performed using metagenome DNA as the template. The amplified PCR products were cloned using TA cloning strategy. Gene sequences were confirmed using Sanger sequencing and submitted to NCBI database. Recombinant proteins of these cellulase genes will be produced to select for highly effective enzymes for biomass degradation. Outcomes from this project will increase the enzyme species for producing value-added products from cellulosic biomass materials.

Subject Area

Molecular biology|Genetics|Animal sciences

Recommended Citation

Santosh Thapa, "Cloning of genes encoding for cellulolytic enzymes for metagenome in goat's rumen" (2015). ETD Collection for Tennessee State University. Paper AAI1599543.