Overexpression of a Fungal beta-Mannanase from Bispora sp MEY-1 in Maize Seeds and Enzyme Characterization
文献类型： 外文期刊
作者： Xu, Xiaolu 1 ; Zhang, Yuhong 2 ; Meng, Qingchang 3 ; Meng, Kun 1 ; Zhang, Wei 2 ; Zhou, Xiaojin 2 ; Luo, Huiying 1 ; Chen 1 ;
作者机构： 1.Chinese Acad Agr Sci, Minist Agr, Feed Res Inst, Key Lab Feed Biotechnol, Beijing 100193, Peoples R China
2.Chinese Acad Agr Sci, Biotechnol Res Inst, 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： 2013 年 8 卷 2 期
页码：
收录情况： SCI
摘要： Background: Mannans and heteromannans are widespread in plants cell walls and are well-known as anti-nutritional factors in animal feed. To remove these factors, it is common practice to incorporate endo-beta-mannanase into feed for efficient nutrition absorption. The objective of this study was to overexpress a beta-mannanase gene directly in maize, the main ingredient of animal feed, to simplify the process of feed production. Methodology/Principal Findings: The man5A gene encoding an excellent beta-mannanase from acidophilic Bispora sp. MEY-1 was selected for heterologous overexpression. Expression of the modified gene (man5As) was driven by the embryo-specific promoter ZM-leg1A, and the transgene was transferred to three generations by backcrossing with commercial inbred Zheng58. Its exogenous integration into the maize embryonic genome and tissue specific expression in seeds were confirmed by PCR and Southern blot and Western blot analysis, respectively. Transgenic plants at BC3 generation showed agronomic traits statistically similar to Zheng58 except for less plant height (154.0 cm vs 158.3 cm). The expression level of MAN5AS reached up to 26,860 units per kilogram of maize seeds. Compared with its counterpart produced in Pichia pastoris, seed-derived MAN5AS had higher temperature optimum (90 degrees C), and remained more beta-mannanase activities after pelleting at 80 degrees C, 100 degrees C or 120 degrees C. Conclusion/Significance: This study shows the genetically stable overexpression of a fungal beta-mannanase in maize and offers an effective and economic approach for transgene containment in maize for direct utilization without any purification or supplementation procedures.
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