Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis)
Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identif...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
KeAi Communications Co., Ltd.
2016-04-01
|
| Series: | Journal of Integrative Agriculture |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311915611443 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1839599235530489856 |
|---|---|
| author | Bi-feng LI Ya-xin ZHU Zhao-bing GU Yuan CHEN Jing LENG Xiao GOU Li FENG Qing LI Dong-mei XI Hua-ming MAO Shu-Li YANG |
| author_facet | Bi-feng LI Ya-xin ZHU Zhao-bing GU Yuan CHEN Jing LENG Xiao GOU Li FENG Qing LI Dong-mei XI Hua-ming MAO Shu-Li YANG |
| author_sort | Bi-feng LI |
| collection | DOAJ |
| description | Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identify specific cellulase in the gayal rumen. A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals. This library contained 38 400 clones with an average insert size of 35.5 kb. The Umcel-1 gene was isolated from this library. Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene, which exhibited the most potent cellulase activity. Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids. The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase (GenBank accession no. YP_004310852.1) from Clostridium lentocellum DSM 5427, with 44% identity and 62% similarity. The Umcel-1 gene was heterologously expressed in Escherichia coli BL21, and recombinant Umcel-1 was purified. The activity of purified recombinant Umcel-1 was assessed, and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45°C. To our knowledge, this study provides the first evidence for a cellulase produced by bacteria in gayal rumen. |
| format | Article |
| id | doaj-art-a00ad11cfc7345b8ac60b26bfc1e8a4c |
| institution | Matheson Library |
| issn | 2095-3119 |
| language | English |
| publishDate | 2016-04-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-a00ad11cfc7345b8ac60b26bfc1e8a4c2025-08-02T15:19:57ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192016-04-01154855861Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis)Bi-feng LI0Ya-xin ZHU1Zhao-bing GU2Yuan CHEN3Jing LENG4Xiao GOU5Li FENG6Qing LI7Dong-mei XI8Hua-ming MAO9Shu-Li YANG10Yunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaInstitute of Microbiology, Chinese Academy of Sciences, Beijing 100101, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaDepartment of Biological Science, Huizhou University, Huizhou 560001, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.ChinaYunnan Provincial Key Laboratory of Animal Nutrition and Feed Science, Yunnan Agricultural University, Kunming 650201, P.R.China; Correspondence YANG Shu-li, Mobile: +86-13759115322Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identify specific cellulase in the gayal rumen. A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals. This library contained 38 400 clones with an average insert size of 35.5 kb. The Umcel-1 gene was isolated from this library. Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene, which exhibited the most potent cellulase activity. Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids. The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase (GenBank accession no. YP_004310852.1) from Clostridium lentocellum DSM 5427, with 44% identity and 62% similarity. The Umcel-1 gene was heterologously expressed in Escherichia coli BL21, and recombinant Umcel-1 was purified. The activity of purified recombinant Umcel-1 was assessed, and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45°C. To our knowledge, this study provides the first evidence for a cellulase produced by bacteria in gayal rumen.http://www.sciencedirect.com/science/article/pii/S2095311915611443gayalruminal microorganismfosmid libraryUmcel-1 genegene expressioncellulase activity |
| spellingShingle | Bi-feng LI Ya-xin ZHU Zhao-bing GU Yuan CHEN Jing LENG Xiao GOU Li FENG Qing LI Dong-mei XI Hua-ming MAO Shu-Li YANG Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis) Journal of Integrative Agriculture gayal ruminal microorganism fosmid library Umcel-1 gene gene expression cellulase activity |
| title | Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis) |
| title_full | Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis) |
| title_fullStr | Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis) |
| title_full_unstemmed | Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis) |
| title_short | Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis) |
| title_sort | screening and characterization of a novel ruminal cellulase gene umcel 1 from a metagenomic library of gayal bos frontalis |
| topic | gayal ruminal microorganism fosmid library Umcel-1 gene gene expression cellulase activity |
| url | http://www.sciencedirect.com/science/article/pii/S2095311915611443 |
| work_keys_str_mv | AT bifengli screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT yaxinzhu screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT zhaobinggu screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT yuanchen screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT jingleng screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT xiaogou screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT lifeng screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT qingli screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT dongmeixi screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT huamingmao screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis AT shuliyang screeningandcharacterizationofanovelruminalcellulasegeneumcel1fromametagenomiclibraryofgayalbosfrontalis |