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A strain of Pichia pastoris capable of degrading citric acid and its application

A Pichia pastoris, citric acid technology, applied in the preparation of fungi, alcoholic beverages, microorganism-based methods, etc., can solve the problems of affecting storage stability, poor tolerance, reducing the flavor and quality of fruit wine, etc. Excellent acid-reducing ability, improved flavor and nutrition, low-cost effect

Active Publication Date: 2020-05-19
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The organic acid in kiwi fruit wine is mainly composed of citric acid. The commonly used acid reduction method in production is chemical method. Although the addition of chemical reagents such as calcium carbonate and potassium bicarbonate has obvious acid reduction effect, it will introduce a large amount of metal ions and reduce the acidity. The flavor and quality of fruit wine will also affect the stability of later storage
In May 2017, Zhang Duo et al. reported a citric acid-degrading strain, Pichia kudriavzevii, which was isolated and purified from citric acid plant wastewater by enrichment and cultivation. Bacterial strain, this bacterial strain can degrade about 84% citric acid in the citric acid medium, but this bacterial strain is very poor to the fruit juice tolerance ability of high sugar, high acid, high alcohol content, thus has limited its application in fruit wine, and The acid-reducing ability of this strain still needs to be further improved

Method used

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  • A strain of Pichia pastoris capable of degrading citric acid and its application
  • A strain of Pichia pastoris capable of degrading citric acid and its application
  • A strain of Pichia pastoris capable of degrading citric acid and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Screening and Identification of Example 1 Bacterial Strains

[0020] (1) Isolation of strains

[0021] Fresh, unpreserved citrus, lemon skin and soil collected from orange groves were used as raw materials, added to a certain volume of normal saline under sterile conditions, and shaken at 26°C and 120r / min in a shaker for 20min. Take 5ml of the supernatant, add it to 100ml of YPD medium containing 1000mg / L streptomycin, and carry out enrichment culture at 28°C, 120r / min shaking culture for 24h, and carry out gradient dilution of the obtained culture solution, the gradient is 10 -1 ~10 -8 , take 1ml of the diluted bacterial solution of each gradient and mix well with the WL nutrient solid medium with citric acid as the only carbon source that has not yet solidified, and after it solidifies, place it upside down in a constant temperature incubator at 28°C for static culture for 3 to 4 days. According to the morphological characteristics of the yeast, suspected colonies ...

Embodiment 2

[0030] The biological acid reduction experiment of embodiment 2 bacterial strains

[0031] Preparation of fermentation broth:

[0032] 4g / L yeast extract powder, 5g / L peptone, 20g / L glucose, 10g / L citric acid, stock solution A40ml / L, stock solution B1ml / L.

[0033] Stock solution A: Potassium dihydrogen phosphate 5.5g, potassium chloride 4.25g, calcium chloride 1.25g, magnesium sulfate 1.25g, dilute to 400ml,

[0034] Stock solution B: 0.25g ferric chloride, 0.25g manganese sulfate, dilute to 100ml.

[0035] Sterilize the fermentation broth at 121°C for 15 minutes for later use; place 1 ml of the frozen JT-1-3 strain thawed on ice into 100 ml of YPD medium, culture at 28°C and shake at 120 r / min for 24 hours to obtain yeast activation liquid, Centrifuge the yeast activation solution at 4000r / min for 20min, wash the obtained cell pellet with sterile physiological saline, then centrifuge the obtained bacterial suspension at 4000r / min for 20min, repeat the above operation until...

Embodiment 3

[0036] Embodiment 3: the biological acid reduction experiment of kiwi fruit wine

[0037] Fresh kiwifruits are selected, washed and peeled, squeezed and added with pectinase for enzymatic hydrolysis, then filtered to obtain the supernatant, pasteurized and set aside. The initial citric acid content measured by high performance liquid chromatography was 12.85g / L.

[0038] Put 1ml of the JT-1-3 freezing solution thawed on ice into 100ml YPD medium, culture at 28°C and shake at 120r / min for 24h to obtain the yeast activation solution, and apply the yeast activation solution to the activation solution at 4000r / min Centrifuge for 20 minutes, use sterile normal saline to wash the obtained bacterial precipitate, then centrifuge the obtained bacterial suspension at 4000r / min for 20 minutes, repeat the above operation until the bacterial precipitate without peculiar smell is obtained, and then wash all the bacterial precipitate into the kiwi fruit In the fruit wine, the inoculum amount ...

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Abstract

The invention discloses a strain of Pichia fermentans strain JT-1-3 capable of degrading citric acid, which is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2018146. The invention also discloses a method for degrading fruit wine citric acid, which comprises inoculating the fruit juice with the Pichia pastoris JT-1-3, and then carrying out fermentation treatment to prepare fruit wine with low citric acid content. The bacterial strain provided by the invention not only has more excellent acid-lowering ability, but also has tolerance to various substances, and can be more widely used in the fermentation of fruit wine for acid-lowering.

Description

technical field [0001] The invention belongs to the field of food, beverage and biotechnology, and relates to a strain of Pichia fermentans strain capable of degrading citric acid, and also relates to the application of the strain in degrading fruit wine citric acid. Background technique [0002] Fresh fruit juices such as citrus, kiwi and blueberry are fermented into fruit wine, which not only retains the unique aroma of the fruit to a certain extent, but also retains the organic acids, vitamins, flavonoids and polyphenols in the fruit to a large extent. This active ingredient has health functions such as increasing appetite, promoting digestion, anti-oxidation and improving cardiovascular and cerebrovascular functions, and can avoid waste caused by untimely consumption of fresh fruits. However, due to the higher organic acid content therein, the sour taste of the fruit wine will be too heavy, and the juice will lose its luster and turbidity, which reduces consumers' desire...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12G3/024C12R1/84
CPCC12G3/02C12N1/165C12R2001/84
Inventor 李二虎钟武潘思轶
Owner HUAZHONG AGRI UNIV