Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells

doi:10.1016/S2095-3119(14)60788-7

Journal of Integrative Agriculture ›› 2015, Vol. 14 ›› Issue (4): 732-740.DOI: 10.1016/S2095-3119(14)60788-7

Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells

ZHANG Xian-wei, LI Zi-cong, MENG Fan-ming, WANG De-hua, LIU De-wu, HE Xiao-yan, SUN Yue, BAI Yin-shan, WU Zhen-fang

1、National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642,P.R.China
2、Wen’s Research Institute, Guangdong Wen’s Food Group Co. Ltd., Yunfu 527439, P.R.China

收稿日期:2014-02-12 出版日期:2015-04-01 发布日期:2015-04-10
通讯作者: WU Zhen-fang,Tel: +86-20-85280369, E-mail: wzfemail@163.com
作者简介:ZHANG Xian-wei, Tel: +86-766-2986345,E-mail: zxianw@163.com;* These authors contributed equally to this study.
基金资助:
This work was funded by a grant from the National Science and Technology Major Projects of China (2014ZX08006004) and three grants from the Department of Science and Technology of Guangdong, China (20111090700016, 2011A020102003 and 2011A020201009).

Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells

ZHANG Xian-wei, LI Zi-cong, MENG Fan-ming, WANG De-hua, LIU De-wu, HE Xiao-yan, SUN Yue, BAI Yin-shan, WU Zhen-fang

1、National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642,P.R.China
2、Wen’s Research Institute, Guangdong Wen’s Food Group Co. Ltd., Yunfu 527439, P.R.China

Received:2014-02-12 Online:2015-04-01 Published:2015-04-10
Contact: WU Zhen-fang,Tel: +86-20-85280369, E-mail: wzfemail@163.com
About author:ZHANG Xian-wei, Tel: +86-766-2986345,E-mail: zxianw@163.com;* These authors contributed equally to this study.
Supported by:
This work was funded by a grant from the National Science and Technology Major Projects of China (2014ZX08006004) and three grants from the Department of Science and Technology of Guangdong, China (20111090700016, 2011A020102003 and 2011A020201009).

摘要/Abstract

摘要： 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 arti?cial 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.

关键词: β-glucanase , bicistronic , pig , feed digestibility , salivary gland cells , transgenic

Abstract: 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 arti?cial 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.

Key words: β-glucanase , bicistronic , pig , feed digestibility , salivary gland cells , transgenic

ZHANG Xian-wei, LI Zi-cong, MENG Fan-ming, WANG De-hua, LIU De-wu, HE Xiao-yan, SUN Yue, BAI Yin-shan, WU Zhen-fang. Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells[J]. Journal of Integrative Agriculture, 2015, 14(4): 732-740.

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Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells

Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells

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