Abstract
Objectives
To analyze the microbial diversity and gene content of a thermophilic cellulose-degrading consortium from hot springs in Xiamen, China using 454 pyrosequencing for discovering cellulolytic enzyme resources.
Results
A thermophilic cellulose-degrading consortium, XM70 that was isolated from a hot spring, used sugarcane bagasse as sole carbon and energy source. DNA sequencing of the XM70 sample resulted in 349,978 reads with an average read length of 380 bases, accounting for 133,896,867 bases of sequence information. The characterization of sequencing reads and assembled contigs revealed that most microbes were derived from four phyla: Geobacillus (Firmicutes), Thermus, Bacillus, and Anoxybacillus. Twenty-eight homologous genes belonging to 15 glycoside hydrolase families were detected, including several cellulase genes. A novel hot spring metagenome-derived thermophilic cellulase was expressed and characterized.
Conclusions
The application value of thermostable sugarcane bagasse-degrading enzymes is shown for production of cellulosic biofuel. The practical power of using a short-read-based metagenomic approach for harvesting novel microbial genes is also demonstrated.
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Acknowledgments
This work was financially supported by the National Natural Science Foundation of China (41306181) and Science Foundation of the Fujian Province, China (2016J06009 & JK2014014). The project was also supported by FAFU grants XJQ201417 and 612014043.
Supporting information
Supplementary Table 1—Homologous genes classified in glycoside hydrolase (GH) families (identity >80 %) from XM70.
Supplementary Table 2—Identities between protein sequence of XM70-cellulase with the several predicted cellulase and peptidase amino acids sequences.
Supplementary Table 3—Similarities between protein sequence of XM70-cellulase with the several predicted cellulase and peptidase amino acids sequences.
Supplementary Fig. 1—Scanning electron micrographs of the sugarcane bagasse. (a) Un-degraded sugarcane bagasse. (b) Surface of the degraded sugarcane bagasse with holes.
Supplementary Fig. 2—Glycoside hydrolase gene families’ distribution. The percentage was calculated according to the contigs aligned number percentage in different GH families. Contigs shared more than 80 % identity with the hit gene.
Supplementary Fig. 3—The protein sequence of XM70- cellulase hit the M42_Frv domain, which is annotated as the “M42 Peptidase, Endoglucanases; Peptidase M42 family, Frv subfamily (cd05656, M42 Peptidase, Endoglucanases)”.
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Zhao, C., Chu, Y., Li, Y. et al. High-throughput pyrosequencing used for the discovery of a novel cellulase from a thermophilic cellulose-degrading microbial consortium. Biotechnol Lett 39, 123–131 (2017). https://doi.org/10.1007/s10529-016-2224-y
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DOI: https://doi.org/10.1007/s10529-016-2224-y