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Alicyclobacillus immunomagnetic microspheres and application thereof

A technology of magnetic microspheres and bacillus, applied in the application, material inspection, food preservation and other directions, can solve the problems of high cost, decreased filtering effect, increased production cost, etc., and achieve the effect of low cost, good effect and fast speed

Active Publication Date: 2014-12-24
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-temperature treatment will have a certain impact on the flavor of the juice, and the membrane filtration method is costly and the filtration effect decreases after a certain period of use, so the filter membrane needs to be replaced periodically according to the filtration effect, resulting in an increase in production costs

Method used

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  • Alicyclobacillus immunomagnetic microspheres and application thereof
  • Alicyclobacillus immunomagnetic microspheres and application thereof
  • Alicyclobacillus immunomagnetic microspheres and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] (1) Silanization reaction of magnetic microspheres

[0045] Dissolve 9.5 g of sodium silicate in 200 mL of deionized water and adjust the pH value to 12-13 with hydrochloric acid, add 5 g of Fe to the sodium silicate solution 3 o 4 Magnetic nucleus ultrasonic treatment for 30 minutes, adjust the pH of the reaction system to 6-7 with hydrochloric acid under the condition of 80-90 ℃ water bath, obtain microspheres by magnetic separation and wash with water to obtain silanized magnetic microspheres with neutral pH;

[0046] (2) Amination modification of magnetic microspheres

[0047] 2 g of silanized magnetic microsphere suspension, 1 mL of deionized water and 10 mL of APTES were added to 150 mL of methanol, after ultrasonic treatment for 30 min, 150 mL of glycerol was added, fully reacted at 85-90 °C for 3 h, and the microspheres were obtained by magnetic separation. And washed three times with methanol and deionized water respectively, freeze-dried to obtain aminated m...

Embodiment 2

[0053] This example differs from example 1 in that the Fe used in this example 3 o 4 The magnetic core is prepared by the following method:

[0054] 3.4 g of FeCl 3 ·6H 2 O with 1.2g FeCl 2 4H 2 O was added to 800 mL of deionized water, and 4 g of PEG-4000 was added, stirred and dissolved under the protection of nitrogen, and ammonia solution was added dropwise to make the pH of the reaction system greater than 10; after that, the reaction system was fully matured in a water bath at 75-85 °C for 20- 40 min, magnetically separated to obtain microspheres and washed with water to obtain Fe 3 o 4 Magnetic nucleus; the Fe was detected 3 o 4 The magnetic core particle size is 15-25 nm, and the magnetic saturation magnetization is 72.18 emu / g.

Embodiment 3

[0056] The difference between this example and Example 1 is that the immunomagnetic microspheres of this example are blocked by a blocking solution: a mixed system containing 5 g BSA and 0.05 ml Tween-20 per 100 ml of PBS is used as the blocking solution, and the adsorption antibody is added The immunomagnetic microspheres were reacted at 37°C for 60 min, and then magnetically separated and fully washed to obtain the immunomagnetic microspheres after blocking treatment.

[0057] One, utilize the immunomagnetic microsphere prepared in embodiment 3 to separate Alicyclobacillus:

[0058] The specific operation steps are: prepare 10 mL of each concentration gradient sample shown in Table 1, add 20 mg immunomagnetic microspheres to each sample to separate and remove the target bacteria: 120-180 rpm, reaction adsorption time 60 min; Isolate the immunomagnetic microbeads that adsorb bacteria, wash the immunomagnetic microbeads with sterile water and suspend them in 200 μL of PBS buff...

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Abstract

The invention discloses alicyclobacillus immunomagnetic microspheres and an application thereof. The disclosed alicyclobacillus immunomagnetic microspheres are obtained by coating aminated magnetic microspheres with oxidized alicyclobacillus specific antibodies. A preparation method of the oxidized alicyclobacillus specific antibodies comprises the following steps: alicyclobacillus specific antibodies are oxidized by using a PBS solution of sodium persulfate, and then the oxidized alicyclobacillus specific antibodies are collected to obtain. The disclosed alicyclobacillus immunomagnetic microspheres can be used for effectively separating alicyclobacillus in fruit juice especially in apple juice.

Description

technical field [0001] The invention belongs to the technical field of food microorganism detection, and in particular relates to immunomagnetic microspheres of Alicyclobacillus and applications thereof. Background technique [0002] Alicyclobacillus, commonly known as acidophilic and thermostable bacteria, belongs to Gram-positive bacillus, rod-shaped, spore-forming, and strictly aerobic. It can survive and grow in a wide temperature range of pH 2.5-6.0, 20-60 ℃, is a common contaminating bacteria in apple and other fruit juices. Due to its heat resistance and acid resistance, ordinary pasteurization is difficult to kill it. The bacterium does not multiply in concentrated fruit juice, but when the concentrated fruit juice is diluted into low-concentration fruit juice, it can metabolize and produce substances with unpleasant flavors - guaiacol and 2,6-dibromophenol, which make the taste and flavor of the juice worse , forming a white precipitate or foggy turbidity, which ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68A23L3/3526
Inventor 岳田利王周利袁亚宏蔡瑞牛晨郭彩霞
Owner NORTHWEST A & F UNIV
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