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A kind of strain and method for degrading naringin

A technology of naringin and strains, applied in the field of degrading naringin, can solve the problem of high cost, and achieve the effects of simple nutritional requirements and cost reduction.

Inactive Publication Date: 2019-04-26
方柏山
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The physical adsorption method will also absorb the nutrients in the fruit juice while absorbing the bitter substance. The food additive method just masks the bitter substance, but does not remove it; and the biological enzyme preparation method, Chinese patent 201010219099. December 1, 2010) disclosed a method for fermenting and producing pomelo juice debitterase, which requires separation and purification of the enzyme, and the cost of this process is relatively high

Method used

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  • A kind of strain and method for degrading naringin
  • A kind of strain and method for degrading naringin

Examples

Experimental program
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Effect test

Embodiment 1

[0034] (1) Isolation and purification of strains

[0035] Take 1g of fresh soil sample from pomelo orchard and add it to 100mL enrichment medium for 48 hours, then take 1mL of enrichment medium and transfer it to a new enrichment medium for continuous enrichment culture for 3 generations. Take 1mL of the enriched culture solution, perform serial dilutions with a 10-fold gradient, and select an appropriate dilution (10 -1 , 10 -5 , 10 -6 ) bacterial suspension 0.2mL, spread in the isolation medium, place in a 30°C incubator and cultivate for 72 hours, pick all the growing strains and separate them by streaking. Continuously streaked and cultured for 3 generations to obtain purified strains. Transfer these pure bacterial strains to liquid medium, 30 ℃, 150rpm, shaker culture, after cultivating 72h, measure the change situation of naringin content according to the following method:

[0036] Determination by high performance liquid chromatography, the conditions are: mobile ph...

Embodiment 2

[0046] Determination of growth curve and degradation of naringin.

[0047] Strain: Phyllobacterium sp. Wb51b CGMCC No. 6472.

[0048] Medium:

[0049] Seed medium (g / L): glucose 1; yeast powder 2; peptone 0.5; naringin 0.5; pH 3.0-5.0, 0.1MPa, sterilized for 20 minutes.

[0050] Fermentation medium (g / L): naringin 2; ammonium sulfate 1; dipotassium hydrogen phosphate 1; anhydrous calcium chloride 0.2; magnesium sulfate 0.5; pH 3.0-6.0, 0.1MPa, sterilization for 20 minutes.

[0051] Seed culture: Inoculate Phyllobacterium sp.Wb51b CGMCC No.6472 into a medium containing 0.5g / L naringin (250mL Erlenmeyer flask, liquid volume 100mL), temperature 30°C, rotation speed 150rpm, shaker culture for 24h.

[0052] Fermentation culture: Inoculate the Phyllobacterium sp.Wb51b CGMCC No.6472 strain after seed culture culture into the fermentation medium, the culture conditions are the same as the seed culture, and samples are taken every 4h to measure the biomass OD respectively 600nm and ...

Embodiment 3

[0057] Degradation of naringin by free cells

[0058] Strain: Phyllobacterium sp. Wb51b CGMCC No. 6472.

[0059] Medium (g / L): naringin 2; yeast extract 0.1; ammonium sulfate 1: calcium chloride 0.2; magnesium sulfate 0.5; dipotassium hydrogen phosphate 1;

[0060] Phyllobacterium sp.Wb51b CGMCC No.6472 was inoculated into the seed medium at a temperature of 30° C. and a rotational speed of 150 rpm, and cultured on a shaker for 72 hours.

[0061] Isolation of free cells:

[0062] The Phyllobacterium sp.Wb51b CGMCC No.6472 cells cultured for 72 hours were collected and centrifuged for 10 minutes in a centrifuge with a temperature of 4° C., a rotation speed of 8000 rpm, and a capacity of 50 mL. Discard the supernatant, wash three times with PBS buffer with pH 6.0 to remove naringin contained in the cells, and finally suspend the cells with PBS buffer with pH 6.0, and store them in a refrigerator at 4°C for later use.

[0063] Under the conditions of naringin concentration of ...

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Abstract

The invention relates to a microbial strain for degrading naringin and a method, and provides Wb51b (phyllobacterium sp.) capable of degrading the naringin. Under certain conditions, 2g / L of naringin serves as an unique carbon source and is used for culturing a microbial strain for 72 hours, concentration of thallus reaches 0.45 g / L, the content of the naringin is reduced to be 0.0037 g / L, and the naringin is nearly degraded completely. 0.6 g / L of naringin is directly degraded by free cells, reaction time is 30 minutes, the content of the naringin is reduced to be 0.1 g / L, and degradation rate reaches 83.3%; and 1.2 g / L of naringin is degraded by immobilized cells, reaction time is 60 minutes, the content of the naringin is reduced to be 0.076 g / L, and the degradation rate reaches 93.7%. The microbial strain can be used for degrading the naringin effectively, nutrition requirements are simple, and the microbial strain is easy to culture. In addition, the microbial strain does not have pathogenicity, and can be directly used in the food industry.

Description

technical field [0001] The invention relates to a leaf bacterium, in particular to a method for degrading naringin. Background technique [0002] Naringenin (Naringenine-7-rhamnosidoglucoside), also known as naringin, citrusin, and isohesperidin, is the most important bitter substance in pomelo peel. Reducing the content of naringin and establishing the industrialization technology of pomelo juice debittering will bring significant progress to the beverage industry. In recent years, the method of microbial degradation of naringin has attracted more and more attention. The types of microorganisms involved include fungi, bacteria and yeast, etc. Among them, fungi have achieved certain results by producing naringinase to degrade naringin; There are few studies on naringin. [0003] As the planting area of ​​honey pomelo continues to expand, the output of honey pomelo has increased year by year. The annual output value of honey pomelo in Fujian Province alone has reached 60 mi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/01
Inventor 方柏山
Owner 方柏山
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