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

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 databas...

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Autores principales: Wang, Tzi-Yuan, Chen, Hsin-Liang, Lu, Mei-Yeh J, Chen, Yo-Chia, Sung, Huang-Mo, Mao, Chi-Tang, Cho, Hsing-Yi, Ke, Huei-Mien, Hwa, Teh-Yang, Ruan, Sz-Kai, Hung, Kuo-Yen, Chen, Chih-Kuan, Li, Jeng-Yi, Wu, Yueh-Chin, Chen, Yu-Hsiang, Chou, Shao-Pei, Tsai, Ya-Wen, Chu, Te-Chin, Shih, Chun-Chieh A, Li, Wen-Hsiung, Shih, Ming-Che
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3177772/
https://www.ncbi.nlm.nih.gov/pubmed/21849025
http://dx.doi.org/10.1186/1754-6834-4-24
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author Wang, Tzi-Yuan
Chen, Hsin-Liang
Lu, Mei-Yeh J
Chen, Yo-Chia
Sung, Huang-Mo
Mao, Chi-Tang
Cho, Hsing-Yi
Ke, Huei-Mien
Hwa, Teh-Yang
Ruan, Sz-Kai
Hung, Kuo-Yen
Chen, Chih-Kuan
Li, Jeng-Yi
Wu, Yueh-Chin
Chen, Yu-Hsiang
Chou, Shao-Pei
Tsai, Ya-Wen
Chu, Te-Chin
Shih, Chun-Chieh A
Li, Wen-Hsiung
Shih, Ming-Che
author_facet Wang, Tzi-Yuan
Chen, Hsin-Liang
Lu, Mei-Yeh J
Chen, Yo-Chia
Sung, Huang-Mo
Mao, Chi-Tang
Cho, Hsing-Yi
Ke, Huei-Mien
Hwa, Teh-Yang
Ruan, Sz-Kai
Hung, Kuo-Yen
Chen, Chih-Kuan
Li, Jeng-Yi
Wu, Yueh-Chin
Chen, Yu-Hsiang
Chou, Shao-Pei
Tsai, Ya-Wen
Chu, Te-Chin
Shih, Chun-Chieh A
Li, Wen-Hsiung
Shih, Ming-Che
author_sort Wang, Tzi-Yuan
collection PubMed
description 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: β-glucosidase, endo-1,4-β-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 β-glucosidase (W5-16143) and one exocellulase (W5-CAT26) showed strong activities and could potentially be developed into commercial enzymes.
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spelling pubmed-31777722011-09-22 Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses Wang, Tzi-Yuan Chen, Hsin-Liang Lu, Mei-Yeh J Chen, Yo-Chia Sung, Huang-Mo Mao, Chi-Tang Cho, Hsing-Yi Ke, Huei-Mien Hwa, Teh-Yang Ruan, Sz-Kai Hung, Kuo-Yen Chen, Chih-Kuan Li, Jeng-Yi Wu, Yueh-Chin Chen, Yu-Hsiang Chou, Shao-Pei Tsai, Ya-Wen Chu, Te-Chin Shih, Chun-Chieh A Li, Wen-Hsiung Shih, Ming-Che Biotechnol Biofuels Research 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: β-glucosidase, endo-1,4-β-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 β-glucosidase (W5-16143) and one exocellulase (W5-CAT26) showed strong activities and could potentially be developed into commercial enzymes. BioMed Central 2011-08-17 /pmc/articles/PMC3177772/ /pubmed/21849025 http://dx.doi.org/10.1186/1754-6834-4-24 Text en Copyright © 2011 Wang et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Wang, Tzi-Yuan
Chen, Hsin-Liang
Lu, Mei-Yeh J
Chen, Yo-Chia
Sung, Huang-Mo
Mao, Chi-Tang
Cho, Hsing-Yi
Ke, Huei-Mien
Hwa, Teh-Yang
Ruan, Sz-Kai
Hung, Kuo-Yen
Chen, Chih-Kuan
Li, Jeng-Yi
Wu, Yueh-Chin
Chen, Yu-Hsiang
Chou, Shao-Pei
Tsai, Ya-Wen
Chu, Te-Chin
Shih, Chun-Chieh A
Li, Wen-Hsiung
Shih, Ming-Che
Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses
title Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses
title_full Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses
title_fullStr Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses
title_full_unstemmed Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses
title_short Functional characterization of cellulases identified from the cow rumen fungus Neocallimastix patriciarum W5 by transcriptomic and secretomic analyses
title_sort functional characterization of cellulases identified from the cow rumen fungus neocallimastix patriciarum w5 by transcriptomic and secretomic analyses
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3177772/
https://www.ncbi.nlm.nih.gov/pubmed/21849025
http://dx.doi.org/10.1186/1754-6834-4-24
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