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Production of a Highly Protease-Resistant Fungal alpha-Galactosidase in Transgenic Maize Seeds for Simplified Feed Processing

文献类型： 外文期刊

作者： Yang, Wenxia 1 ; Zhang, Yuhong 2 ; Zhou, Xiaojin 2 ; Zhang, Wei 2 ; Xu, Xiaolu 1 ; Chen, Rumei 2 ; Meng, Qingchang 3 ; Yu 1 ;

作者机构： 1.Chinese Acad Agr Sci, Feed Res Inst, Beijing 100193, Peoples R China

2.Chinese Acad Agr Sci, Inst Biotechnol, Beijing 100193, Peoples R China

3.Jiangsu Acad Agr Sci, Inst Food Crops, Nanjing, Jiangsu, Peoples R China

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2015 年 10 卷 6 期

页码：

收录情况： SCI

摘要： Raffinose-family oligosaccharide (RFO) in soybeans is one of the major anti-nutritional factors for poultry and livestocks. alpha-Galactosidase is commonly supplemented into the animal feed to hydrolyze alpha-1,6-galactosidic bonds on the RFOs. To simplify the feed processing, a protease-resistant alpha-galactosidase encoding gene from Gibberella sp. strain F75, aga-F75, was modified by codon optimization and heterologously expressed in the embryos of transgentic maize driven by the embryo-specific promoter ZM-leg1A. The progenies were produced by backcrossing with the commercial inbred variety Zheng58. PCR, southern blot and western blot analysis confirmed the stable integration and tissue specific expression of the modified gene, aga-F75m, in seeds over four generations. The expression level of Aga-F75M reached up to 10,000 units per kilogram of maize seeds. In comparison with its counterpart produced in Pichia pastoris strain GS115, maize seed-derived Aga-F75M showed a lower temperature optimum (50 degrees C) and lower stability over alkaline pH range, but better thermal stability at 60 degrees C to 70 degrees C and resistance to feed pelleting inactivation (80 degrees C). This is the first report of producing alpha-galactosidase in transgenic plant. The study offers an effective and economic approach for direct utilization of alpha-galactosidase-producing maize without any purification or supplementation procedures in the feed processing.

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