Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells
Many animal feed grains contain high β-glucan in the cell wall. Pigs do not secret β-glucanase to degrade the β-glucan in their feed. The indigestible β-glucan not only blocks the release of nutrients from the grain cell wall, but also increases the digesta viscosity in the gastrointestinal tract of...
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KeAi Communications Co., Ltd.
2015-04-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311914607887 |
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| author | Xian-wei ZHANG Zi-cong LI Fan-ming MENG De-hua WANG De-wu LIU Xiao-yan HE Yue SUN Yin-shan BAI Zhen-fang WU |
| author_facet | Xian-wei ZHANG Zi-cong LI Fan-ming MENG De-hua WANG De-wu LIU Xiao-yan HE Yue SUN Yin-shan BAI Zhen-fang WU |
| author_sort | Xian-wei ZHANG |
| collection | DOAJ |
| description | Many animal feed grains contain high β-glucan in the cell wall. Pigs do not secret β-glucanase to degrade the β-glucan in their feed. The indigestible β-glucan not only blocks the release of nutrients from the grain cell wall, but also increases the digesta viscosity in the gastrointestinal tract of pigs. Therefore, dietary β-glucan significantly inhibits nutrient digestion and absorption in pigs. Transgenic expression of β-glucanase in the digestive tract of pigs may offer a solution to solve this problem. In the current study, four artificial codon-optimized β-glucanases genes was prepared and expressed in porcine cells. Only pBgA and pEgx showed high activity in transfected pig kidney cells. To improve the pH range and pH stability of β-glucanase, the two β-glucanases, pBgA and pEgx, were co-expressed in pig kidney cells and salivary gland cells by Linker A3 or 2A peptide. The resulting dual enzymes of pBgA3pEg and pBg2ApEg showed significantly enlarged pH range and significantly increased pH stability, as compared to parental enzymes. These results provide useful data for future study on increasing the feed digestibility of pigs by transgenic expression of β-glucanase in their salivary glands. |
| format | Article |
| id | doaj-art-bfddd1251fc340a2a99a4bf664d47f13 |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2015-04-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-bfddd1251fc340a2a99a4bf664d47f132025-08-20T03:57:39ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192015-04-0114473274010.1016/S2095-3119(14)60788-7Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cellsXian-wei ZHANG0Zi-cong LI1Fan-ming MENG2De-hua WANG3De-wu LIU4Xiao-yan HE5Yue SUN6Yin-shan BAI7Zhen-fang WU8National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.China; Wen's Research Institute, Guangdong Wen's Food Group Co. Ltd., Yunfu 527439, P.R.China; ZHANG Xian-wei, Tel: +86-766-2986345National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.ChinaNational Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.ChinaNational Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.ChinaNational Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.ChinaNational Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.China; Wen's Research Institute, Guangdong Wen's Food Group Co. Ltd., Yunfu 527439, P.R.ChinaNational Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.ChinaNational Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.ChinaNational Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, P.R.China; Wen's Research Institute, Guangdong Wen's Food Group Co. Ltd., Yunfu 527439, P.R.China; Correspondence WU Zhen-fang, Tel: +86-20-85280369Many animal feed grains contain high β-glucan in the cell wall. Pigs do not secret β-glucanase to degrade the β-glucan in their feed. The indigestible β-glucan not only blocks the release of nutrients from the grain cell wall, but also increases the digesta viscosity in the gastrointestinal tract of pigs. Therefore, dietary β-glucan significantly inhibits nutrient digestion and absorption in pigs. Transgenic expression of β-glucanase in the digestive tract of pigs may offer a solution to solve this problem. In the current study, four artificial codon-optimized β-glucanases genes was prepared and expressed in porcine cells. Only pBgA and pEgx showed high activity in transfected pig kidney cells. To improve the pH range and pH stability of β-glucanase, the two β-glucanases, pBgA and pEgx, were co-expressed in pig kidney cells and salivary gland cells by Linker A3 or 2A peptide. The resulting dual enzymes of pBgA3pEg and pBg2ApEg showed significantly enlarged pH range and significantly increased pH stability, as compared to parental enzymes. These results provide useful data for future study on increasing the feed digestibility of pigs by transgenic expression of β-glucanase in their salivary glands.http://www.sciencedirect.com/science/article/pii/S2095311914607887β-glucanasebicistronicpigfeed digestibilitysalivary gland cellstransgenic |
| spellingShingle | Xian-wei ZHANG Zi-cong LI Fan-ming MENG De-hua WANG De-wu LIU Xiao-yan HE Yue SUN Yin-shan BAI Zhen-fang WU Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells Journal of Integrative Agriculture β-glucanase bicistronic pig feed digestibility salivary gland cells transgenic |
| title | Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells |
| title_full | Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells |
| title_fullStr | Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells |
| title_full_unstemmed | Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells |
| title_short | Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells |
| title_sort | characterization of dual enzyme resulted from bicistronic expression of two β glucanases in porcine cells |
| topic | β-glucanase bicistronic pig feed digestibility salivary gland cells transgenic |
| url | http://www.sciencedirect.com/science/article/pii/S2095311914607887 |
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