Method for removing patulin in orange juice by utilizing immobilized inactivated yeast cells of magnetic microspheres and application of method

A patulin, yeast cell technology, applied in microorganism-based methods, immobilized on or in inorganic carriers, immobilized on/in organic carriers, etc., can solve the use limitations, separation and removal difficulties, limit free The practical application of living biological adsorption method, etc., to achieve the effect of efficient and rapid separation, avoiding adverse effects and low cost

Active Publication Date: 2016-08-10
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing studies have shown that patulin can almost completely disappear in the fermentation process of some living microorganisms. However, living microorganisms will inevitably have a certain impact on fruit juice products and human health, so its use is limited
In addition, due to the small size of microbial cells, it is difficult to completely separate and remove from fruit juice products after adsorbing patulin, which further limits the practical application of free living organism biosorption method in fruit juice production

Method used

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  • Method for removing patulin in orange juice by utilizing immobilized inactivated yeast cells of magnetic microspheres and application of method
  • Method for removing patulin in orange juice by utilizing immobilized inactivated yeast cells of magnetic microspheres and application of method
  • Method for removing patulin in orange juice by utilizing immobilized inactivated yeast cells of magnetic microspheres and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Evaluation of patulin removal ability of 8 strains in simulated acidified water system

[0030] 1. Preparation of main reagents

[0031] CM0005 lactic acid bacteria medium: yeast extract 7.5g, glucose 10.0g, tomato juice 100mL, peptone 7.5g, KH2PO4 2.0g, Tween 80 0.5mL, distilled water 900mL, pH 7.0. Sterilize at 121°C for 15min. C0006MRS medium: casein peptone 10.0g, beef powder 8.0g, yeast powder 4.0g, glucose 20g, magnesium sulfate 0.2g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g , Tween 80 1.0g, distilled water 1000mL, pH 6.2±0.2. Sterilize at 121°C for 15min. CM0181 yeast culture medium: yeast extract 3.0g, malt extract 3.0g, peptone 5.0g, glucose 10.0g, agar 20.0g, distilled water 1000mL. Sterilize at 115°C for 25 minutes.

[0032] CM0221 tryptone-soytone agar: tryptone-soytone agar 40.0g, distilled water 1000mL. Sterilize at 121°C for 15 minutes.

[0033] CM0787 enhanced Clostridium ...

Embodiment 2

[0057] Example 2: Evaluation of 3 kinds of preferred bacterial strains for free inactivation on the removal ability of patulin in orange juice

[0058] Take the three preferred bacteria in the -80°C glycerol stock solution with 2% inoculum size, activate and culture them in their corresponding medium for 20 hours, and then passage twice with 2% inoculum size, centrifuge for 10 min (5000r / min, 4°C) The cells were collected, washed with distilled water and centrifuged three times, sterilized at 121°C for 15 minutes, and then freeze-dried. Add 0.2 g of inactivated bacteria powder to 10 mL of orange juice system containing 1000 μg / L PAT, and then place it in a constant temperature shaker at 10 °C for 10 h at a speed of 120 r / min. After the treatment, the adsorption system solution was centrifuged using a high-speed refrigerated centrifuge (13000r / min, 10min, 4°C), and the residual amount of patulin in the supernatant was determined by HPLC to evaluate the adsorption efficiency.

...

Embodiment 3

[0060] Example 3: Analysis of Candida.utilis CICC1769 Morphological Characteristics

[0061] Insert the optimal adsorption strain Candida.utilis CICC1769 into the corresponding liquid medium for continuous activation for 3 generations, separate it by streaking in the solid medium, place it in a constant temperature incubator at 30°C for 36-48 hours, and observe the colony shape. At the same time, a single colony was picked for Gram staining, and the colony shape and individual shape were observed under a microscope. The results were as follows: Figure 4 Shown:

[0062] Candida.utilis CICC1769 colony morphology: Cultured at 30°C for 48 hours, it grows well on CM0181 yeast solid medium, the colonies are milky white, smooth, shiny, with neat edges. Colony individuals such as Figure 4 As shown, the cell shape is sausage-shaped, and the bacteria exist in single, paired or aggregated chains, with a width of about 4 μm and a length of about 10 μm, with smooth ends, Gram-positive,...

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Abstract

The invention relates to a method for removing patulin in orange juice by utilizing immobilized inactivated yeast cells of magnetic microspheres and application of the method. The method comprises the steps: adopting an inverse suspension crosslinking method, by means of glutaraldehyde crosslinking, polymerizing with chitosan molecules to prepare magnetic Fe3O4 / CTS microspheres, and optimizing the process parameters of immobilized inactivated Candida. u CICC1769 cells of the magnetic Fe3O4 / CTS microspheres, thereby utilizing the immobilized inactivated yeast cells of the magnetic microspheres to remove patulin in the orange juice. The method has the advantages of good adsorption effect, low cost and environmental protection, the efficient and rapid separation of the inactivated yeast cells and a target product can also be realized, and various adverse effects of living microorganisms on the product and the human body are simultaneously avoided.

Description

technical field [0001] The invention belongs to the technical field of food safety, and in particular relates to a method for removing patulin in citrus juice by using magnetic microspheres to immobilize inactivated yeast cells and an application thereof. Background technique [0002] PAT (patulin, PAT), also known as patulin, is a hemiacetal lactone with a molecular formula of C 7 h 6 o 4 , the chemical name is 4-hydroxy-4-hydro-furan-[3,2 carbon]-pyran-2(6 hydrogen)one. PAT has acute toxicity, including damage to the lungs, brain edema, liver, spleen, kidney and immune system of animals; it also has chronic toxicity, manifested in cytotoxicity, genotoxicity and immunotoxicity to animals. It has a strong affinity for sulfhydryl groups and can react with sulfhydryl-containing proteins and polypeptides to destroy single- and double-strand DNA, thereby inhibiting the synthesis of DNA and RNA, and can also inhibit the activities of various enzymes, especially enzymes in some...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10A23L2/80C12R1/72
CPCA23L2/80C12N11/10C12N11/14
Inventor 彭帮柱潘思轶徐晓云薛淑静关键范刚胡昊
Owner HUAZHONG AGRI UNIV
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