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Production of two Highly Active Bacterial Phytases with Broad pH Optima in Germinated Transgenic Rice Seeds

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Abstract

Phytate is the main storage form of phosphorus in many plant seeds, but phosphate bound in this form is not available to monogastric animals. Phytase, an enzyme that hydrolyzes phosphate from phytate, has the potential to enhance phosphorus availability in animal diets when engineered in rice seeds as a feed additive. Two genes, derived from a ruminal bacterium Selenomonas ruminantium (SrPf6) and Escherichia coli (appA), encoding highly active phytases were expressed in germinated transgenic rice seeds. Phytase expression was controlled by a germination inducible α-amylase gene (αAmy8) promoter, and extracellular phytase secretion directed by an αAmy8 signal peptide sequence. The two phytases were expressed in germinated transgenic rice seeds transiently and in a temporally controlled and tissue-specific manner. No adverse effect on plant development or seed formation was observed. Up to 0.6 and 1.4 U of phytase activity per mg of total extracted cellular proteins were obtained in germinated transgenic rice seeds expressing appA and SrPf6 phytases, respectively, which represent 46–60 times of phytase activities compared to the non-transformant. The appA and SrPf6 phytases produced in germinated transgenic rice seeds had high activity over broad pH ranges of 3.0–5.5 and 2.0–6.0, respectively. Phytase levels and inheritance of transgenes in one highly expressing plant were stable over four generations. Germinated transgenic rice seeds, which produce a highly active recombinant phytase and are rich in hydrolytic enzymes, nutrients and minerals, could potentially be an ideal feed additive for improving the phytate-phosphorus digestibility in monogastric animals.

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Authors and Affiliations

  1. Institute of Molecular Biology, Academia Sinica, Nankang, Taipei, 11529, Taiwan, ROC

    Chwan-Yang Hong & Su-May Yu

  2. Institute of BioAgricultural Sciences, Academia Sinica, Nankang, Taipei, 11529, Taiwan, ROC

    Kuo-Joan Cheng

  3. Taiwan Agricultural Research Institute, Wu-Fong, Taichung, Taiwan, ROC

    Tung-Hai Tseng & Chang-Sheng Wang

  4. Department of Agronomy, National Taiwan University, Taipei, Taiwan, ROC

    Chwan-Yang Hong & Li-Fei Liu

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  1. Chwan-Yang Hong
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  2. Kuo-Joan Cheng
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  3. Tung-Hai Tseng
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  4. Chang-Sheng Wang
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  5. Li-Fei Liu
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  6. Su-May Yu
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Correspondence to Li-Fei Liu.

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Hong, CY., Cheng, KJ., Tseng, TH. et al. Production of two Highly Active Bacterial Phytases with Broad pH Optima in Germinated Transgenic Rice Seeds. Transgenic Res 13, 29–39 (2004). https://doi.org/10.1023/B:TRAG.0000017158.96765.67

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  • DOI: https://doi.org/10.1023/B:TRAG.0000017158.96765.67

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