Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses

Tzi Yuan Wang, Hsin Liang Chen, Mei Yeh J. Lu, Yo Chia Chen, Huang Mo Sung, Chi Tang Mao, Hsing Yi Cho, Huei Mien Ke, Teh Yang Hwa, Sz Kai Ruan, Kuo Yen Hung, Chih Kuan Chen, Jeng Yi Li, Yueh Chin Wu, Yu Hsiang Chen, Shao Pei Chou, Ya Wen Tsai, Te Chin Chu, Chun Chieh A. Shih, Wen Hsiung LiMing Che Shih

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Abstract

Background: Neocallimastix patriciarum is one of the common anaerobic fungi in the digestive tracts of ruminants that can actively digest cellulosic materials, and its cellulases have great potential for hydrolyzing cellulosic feedstocks. Due to the difficulty in culture and lack of a genome database, it is not easy to gain a global understanding of the glycosyl hydrolases (GHs) produced by this anaerobic fungus. Results: We have developed an efficient platform that uses a combination of transcriptomic and proteomic approaches to N. patriciarum to accelerate gene identification, enzyme classification and application in rice straw degradation. By conducting complementary studies of transcriptome (Roche 454 GS and Illumina GA IIx) and secretome (ESI-Trap LC-MS/MS), we identified 219 putative GH contigs and classified them into 25 GH families. The secretome analysis identified four major enzymes involved in rice straw degradation: -glucosidase, endo-1,4 - xylanase, xylanase B and Cel48A exoglucanase. From the sequences of assembled contigs, we cloned 19 putative cellulase genes, including the GH1, GH3, GH5, GH6, GH9, GH18, GH43 and GH48 gene families, which were highly expressed in N. patriciarum cultures grown on different feedstocks. Conclusions: These GH genes were expressed in Pichia pastoris and/or Saccharomyces cerevisiae for functional characterization. At least five novel cellulases displayed cellulytic activity for glucose production. One -glucosidase (W5-16143) and one exocellulase (W5-CAT26) showed strong activities and could potentially be developed into commercial enzymes.

Original languageEnglish
Article number24
JournalBiotechnology for Biofuels
Volume4
DOIs
Publication statusPublished - 2011

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Renewable Energy, Sustainability and the Environment
  • Energy(all)
  • Management, Monitoring, Policy and Law

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    Wang, T. Y., Chen, H. L., Lu, M. Y. J., Chen, Y. C., Sung, H. M., Mao, C. T., Cho, H. Y., Ke, H. M., Hwa, T. Y., Ruan, S. K., Hung, K. Y., Chen, C. K., Li, J. Y., Wu, Y. C., Chen, Y. H., Chou, S. P., Tsai, Y. W., Chu, T. C., Shih, C. C. A., ... Shih, M. C. (2011). Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses. Biotechnology for Biofuels, 4, [24]. https://doi.org/10.1186/1754-6834-4-24