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Study on the Detection Method of the Phytase Activity in Transgenic Corn

Author: LiuHongMin
Tutor: LiJing
School: Jilin University
Course: Occupational and Environmental Health
Keywords: Phytase Genetically modified corn Detection methods Orthogonal test
CLC: S513
Type: Master's thesis
Year: 2011
Downloads: 110
Quote: 1
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Abstract


Phytase (Phytase) catalytic phytic acid and phytic acid saline solution into inositol phosphate (or phosphate) the general term for a class of enzymes, is a phosphate monoester hydrolase. Phytase can decompose the phytic acid, the release of a large amount of inorganic phosphorus, phytase, if added in the feed can be the decomposition of large amounts of phosphorus, monogastric animals phytic acid to enhance the absorption rate of the organic phosphorus, reducing the discharge of phosphorus , reducing phosphorus caused environmental pollution. Plant as phytase reactor has become a research hotspot. Corresponding national standards exist for dietary phytase activity detection method of, but less phytase activity assay method reported in the plant, therefore developed scientific and practical determination of plant phytase extraction and the determination of the method has practical significance. Objective: In this paper, to optimize the extraction conditions and testing conditions transgenic maize phytase to determine the best method for extraction and activity determination of phytase in corn and phytase gene in Daejeon and potted phytase activity in corn dynamic monitoring, to provide a basis for the evaluation of genetically modified corn phytase activity. Method: extraction method using ultrasonic and shock conditions affect phytase extracted, such as solid-liquid ratio, extraction time, extract the pH univariate analysis, selected in the univariate analysis phytase activity is relatively high the level of experimental factors, L9 (33) orthogonal table to optimize the extraction conditions of phytase. Spectrophotometric determination of the impact of phytase conditions, such as reaction time, substrate concentration, buffer pH univariate analysis, elected phytase activity relatively high level of experimental factors in the univariate analysis, the use of L9 (33) orthogonal table optimize the conditions of phytase Determination. Extraction and measurement methods, the application of this article to determine the different growth stages Daejeon and potted turn phytase maize phytase activity detection, and compare. Results: The extract of genetically modified corn in the phytase trial results show that, when the solid-liquid ratio of 1:4, the extraction time was 15min, the pH of the extract 5.50, the highest phytase activity, the highest extraction rate. Transgenic corn phytase activity measurement conditions of the study results show that the reaction time was 30min, the pH of the buffer solution is 5.50, the substrate concentration was the highest activity of the phytase detection O.10mmol / L. Daejeon and potted plants of different growth stages phytase gene in maize phytase activity compared with the control group, only part of the leaf, stem, root phytase activity is high, but there was no significant difference in the transgenic maize kernels The phytase activity higher than that in the control group. Conclusion: According to the results of this study the following conclusions: 1. Using ultrasonic methods to extract the best conditions for phytase gene in maize phytase solid-liquid ratio of 1:4, the extraction time was 15min, the pH of the extract is 5.50 2. spectrophotometry method to detect the best conditions for phytase in the phytase gene in maize is the reaction time is 30 min, the buffer pH value of 5.50, the substrate concentration for 0.10mmol / L. 3 the Daejeon and potted turn phytase maize only part of leaves, stems, roots phytase activity relative to the control group corn higher grain phytase activity is much higher than corn.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Corn ( maize )
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