Method for evaluating action effect of heat-resistant phytase in vitro

Abstract

The invention belongs to the technical field of enzyme preparation evaluation, and relates to a method for evaluating the action effect of heat-resistant phytase in vitro, which comprises the following steps: measuring the substance amount y0 of total phosphorus in a sample to be acted; adding a heat-resistant phytase solution subjected to high-temperature treatment into the sample to be acted, simulating digestion in a stomach environment, then simulating digestion in an intestine environment, then measuring the substance amount y2 of inorganic phosphorus digested by the heat-resistant phytase in the sample, and measuring the substance amount y1 of the inorganic phosphorus in the sample in the blank control; substituting y0, y2 and y1 into y in the formula I, and respectively calculatingthe total phosphorus content p0 in the sample to be acted, the inorganic phosphorus content p2 of the sample digested by the heat-resistant phytase and the inorganic phosphorus content p1 of the sample in the blank control; and finally, substituting p0, p2 and p1 into a formula II to calculate the phosphorus release rate P of the heat-resistant phytase in the sample, and evaluating the action effect of the heat-resistant phytase on the sample according to the phosphorus release rate P of the heat-resistant phytase in the sample, so that the method has the advantages of simplicity, rapidness, accuracy, stability, good reproducibility and the like.

Description

technical field [0001] The invention belongs to the technical field of enzyme preparation evaluation, and in particular relates to a method for evaluating the effect of heat-resistant phytase in vitro. Background technique [0002] Phytase has been widely used in livestock and poultry aquaculture and other breeding industries. After phytase is added to feed, it must withstand the high temperature treatment in the process of feed processing. After the feed is eaten, phytase needs to enzymatically decompose nutrients Tolerance to different pH environments in livestock and poultry, therefore, the enzymatic hydrolysis effect will be affected by many factors. However, there is still no unified standard for the evaluation method of phytase effect. The biomimetic digestion system is often used, but the evaluation period is still very long when many samples are evaluated. [0003] The invention patent with the publication number CN 106434850 A discloses a method for evaluating the ...

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Problems solved by technology

[0003] The invention patent with the publication number of CN 106434850 A discloses a method for evaluating the activity of high-temperature-resistant phytase, which specifically passes the action of phytase in the simulated gastric environment. The content of water-soluble phosphorus released by the sample is used to evaluate the activity of high-temperature-resistant phytase, but it has the following problems: 1) In this patent, it is believed that the content of water-soluble phosphorus released by high-temperature-resistant phytase in feed samples is greater, It shows that the higher the high temperature resistant phytase activity, but the more water-soluble phosphorus content released, that is, the higher the phosphorus release rate, it can only show that the better the substrate affinity for the evaluation raw material, the better the degradation effect, while the same phytic acid The degradation effect of enzymes on different raw materials is different, which is related to factors such as the total phosphorus content in raw materials and the structure of phytate phosphorus. Therefore, the content of water-soluble phosphorus released from feed samples treated with high-temperature-resistant phytase indicates the presence of high-temperature-resistant phytase activity. Inaccurate, and the patent uses the released phosphorus content to represent the phytase activity, which cannot reflect the influence of the total phosphorus in the raw material on the result; 2) When the enzyme preparation is subjected to high temperature treatment, the patent uses high-pressure sterilization Pot, the temperature is unstable, and it is only suitable for solid enzyme preparations, not for liquid enzyme preparations; 3) This patent only simulates the digestion of phytase in the stomach environment for 2 hours, but in fact, phytase is added to After entering the feed, it is generally digested in the stomach for 45min-1h, and then enters the intestine for 4h. Therefore, there is a certain difference between the simulated digestion and the actual digestion in the in vitro gastric environment, and it cannot accurately reflect the actual digestion.

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