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A kind of Pichia anomaly and its fermentation culture and application

A technology of Pichia anomaly and aquatic animals, applied in the fields of application, biochemical equipment and methods, microorganisms, etc., can solve the problems of unreported application and limited resource mining of Pichia anomaly strains, etc., and increase the nutritional value , Improve the nutritional value and function, and benefit the growth effect

Active Publication Date: 2020-11-06
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Subtropical regions in my country have abundant rainfall, high air humidity, and climate suitable for the growth of environmental microorganisms. Yeast strain resources are abundant, but the resource mining of abnormal Pichia pastoris is limited. Yeast killing systems have been reported, but there is no report on the application of Pichia anomaly liquid and solid-state fermentation cultures in livestock and poultry aquaculture

Method used

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  • A kind of Pichia anomaly and its fermentation culture and application
  • A kind of Pichia anomaly and its fermentation culture and application
  • A kind of Pichia anomaly and its fermentation culture and application

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1: Strain isolation and identification

[0032] Strain isolation

[0033] Take 10g of Anren Folk Sweet Liquor Koji sample in a 250ml Erlenmeyer flask containing 100mL sterile water, shake it on a vortexer and let it stand for a while, take 1mL of bacterial suspension and 9ml of normal saline and mix well, then take 1ml The bacteria suspension is mixed with 9mL of physiological saline, and so on, set different concentration gradients, the gradient is 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , Take 200μl of bacterial suspension and inoculate it on the separation plate in turn, culture it at 28℃ for 2~3d, pick a typical single colony according to the colony morphology (colony morphology: round, hairy surface is not smooth, color is Milky white opaque, raised in the middle, not easy to provoke, size 3mm-5mm), the abnormal Pichia yeast is screened out through microscopic examination.

[0034] PDA medium: 20g potato, 20g glucose, 20g agar, 1000ml water, natural pH, 115℃, steri...

Embodiment 2

[0040] Pichia anomala AR 2016 The large-scale fermentation:

[0041] Pichia anomala AR will be preserved 2016 Inoculate it on the PDA medium, place it in a constant temperature incubator at 28°C for 36 hours, pick out an inoculation loop of Pichia anomala AR 2016 In the seed culture solution (mass fraction (%): glucose 2%, peptone 1%, yeast powder 1%, natural pH, the rest is double-distilled water), cultured on a constant temperature shaker at 28° C. and 200 rpm for 24 hours.

[0042] The volume of the 500L fermentation tank is 65% (V / V), and the liquid fermentation medium includes, (mass fraction, %): glucose 4.0%, peptone 1.5%, yeast powder 2.0%, pH 6.0, 121℃ high temperature steam Disinfect for 20 minutes, cool to 40°C, inoculate 10% of the seed solution with 24 hours of bacterial age, and ferment for 96 hours at 28°C and 200 rpm. The biomass of the fermentation broth is 15g / L and the yield of killer factor is 5.86mg / L. The yield of β-1,3-glucanase was 964 U / L and the yield of p...

Embodiment 3

[0045] Pichia anomala AR 2016 Protein and amino acid content

[0046] The fermentation broth in Example 2 was centrifuged to collect bacterial sludge, and dried at 55° C. to obtain dry bacterial powder.

[0047] Amino acid determination: accurately weigh 120 mg of dry bacteria powder into a test tube, add 10 mL of 16 mol hydrochloric acid to the test tube containing the sample, shake and mix. Use an alcohol blowtorch to thin the lower 1 / 3 of the test tube to 4-6mm, and then seal the tube after vacuuming for 10 minutes. The treated test tube was hydrolyzed in a sand bath at 110°C constant temperature oven for 22 hours, cooled to room temperature, shaken and filtered, and 1mL of the filtrate was placed in a 50mL beaker. The filtrate was evaporated to dryness in a 60°C constant temperature water bath, and 0.02mol hydrochloric acid was added and diluted 4 times. Filter with 0.22μm membrane and measure with Hitachi L-8800 automatic amino acid analyzer. Kjeldahl method (GB5009.5-2003) ...

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Abstract

This article belongs to the field of biotechnology, and discloses a Pichia anomaly and its fermentation culture and application. The Pichia anomaly is: Pichia anomaly AR 2016 , CCTCC NO: M 2017594. Utilize the fermentation medium provided by the present invention to carry out liquid state fermentation to it, the biomass of fermentation broth is 15 g / L, the output of killing factor is 5.86 mg / L, and the output of β-1,3-glucanase is 964 The output of U / L and phytase was 1675 U / L. Pichia anomaly of the present invention has the effect of resisting and inhibiting Escherichia coli, Salmonella pullorum, sensitive C-type Candida albicans, Actinobacillus pleuropneumoniae, Sarcina luteus, Staphylococcus aureus, resistant to gastric acid and bile salts . Therefore, the Pichia anomaly provided by the invention can be used as a feed additive for livestock, poultry and aquatic animals, and has broad application prospects.

Description

Technical field [0001] The present invention relates to the field of biotechnology, and more specifically to an abnormal Pichia yeast and its fermentation culture and application. [0002] technical background [0003] Abnormal Pichia is a basidiomycete, the colony is milky white, it is a simple pseudohyphae, the cell shape is round, and it is the germination of vegetative cells. Pichia anomalous has anti-candida albicans, gastric acid and bile salt resistance, and its liquid and solid fermentation products also contain killer factors, β-1,3-glucanase and phytase. β-1,3-glucanase can catalyze the hydrolysis of β-glucan in grain cell walls. Phytase can hydrolyze phosphoric acid residues from phytic acid, destroying the strong affinity of phytic acid for mineral elements, thereby increasing the nutritional value of mineral elements, and improving the nutritional value of plant feeds. Abnormal Pichia pastoris fermentation produces killer factor, β-1,3-glucanase and phytase. The cult...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16A23K10/18A23K50/75A23K50/80A23K50/30A23K50/60A61K36/062A61P31/04A61P31/10C12R1/84
CPCA61K36/062A23K10/18A23K50/30A23K50/60A23K50/75A23K50/80C12N1/16C12N1/165C12R2001/84Y02A40/818
Inventor 唐志如孙志洪马娅君毛俊霞
Owner SOUTHWEST UNIV
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