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Pichia anomala as well as fermentation culture and application thereof

A technology of Pichia anomaly and a culture medium, applied to Pichia anomaly and its fermentation culture and application fields, can solve the problems of unreported application, limited mining of Pichia anomaly strain resources, etc., to increase nutritional effect. price, improve nutritional value function, beneficial to growth effect

Active Publication Date: 2018-04-20
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Pichia anomala as well as fermentation culture and application thereof
  • Pichia anomala as well as fermentation culture and application thereof
  • Pichia anomala as well as fermentation culture and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: strain isolation and identification

[0031] Strain Isolation

[0032] Take 10g of Anren folk sweet wine koji sample in a 250ml Erlenmeyer flask filled with 100mL of sterile water, oscillate on a vortex instrument and let it stand for a while, take 1mL of bacterial suspension and mix with 9ml of normal saline, and then take 1ml Mix the bacterial suspension with 9mL of normal saline, and so on, set different concentration gradients, the gradient is 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 , inoculate 200 μl of the bacterial suspension on a separation plate in turn, place it at 28°C for 2-3 days, and pick a typical single colony according to the colony shape (colony shape: round, hairy and not smooth on the surface, and the color is Milky white opaque, raised in the middle, not easy to provoke, size 3mm-5mm), abnormal Pichia pastoris was roughly screened out through microscopic examination.

[0033] PDA medium: 20g potato, 20g glucose, 20g agar, 1000ml water,...

Embodiment 2

[0039] Pichia anomala AR 2016 Large-scale fermentation of:

[0040] The preserved Pichia anomala AR 2016 Inoculate on PDA medium, place in a constant temperature incubator at 28°C for 36 hours, and pick an inoculation loop of Pichia anomala AR 2016 In the seed culture solution (mass fraction (%): 2% glucose, 1% peptone, 1% yeast powder, natural pH, the rest is double distilled water), cultivated on a constant temperature shaker at 28°C and 200rpm for 24h.

[0041] The liquid 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, steamed at 121°C Sterilize for 20 minutes, cool down to 40°C, inoculate 10% seed solution with 24h of bacterial age, at 28°C, after 200rpm fermentation for 96h, the biomass of the fermentation liquid is 15g / L, and the output of phagocytic factor is 5.86mg / L. The yield of β-1,3-glucanase was 964U / L and that of phytase was 1675U / L.

[0...

Embodiment 3

[0044] Pichia anomala AR 2016 protein and amino acid content

[0045] The fermentation broth in Example 2 was centrifuged to collect the bacteria sludge, and dried at 55° C. to obtain dry bacteria powder.

[0046] Amino Acid Determination: Accurately weigh 120 mg of dry bacterial 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 4-6 mm, vacuumize for 10 minutes and then seal the tube. The treated test tube was hydrolyzed in a sand bath in a constant temperature oven at 110°C for 22 hours, taken out and cooled to room temperature, shaken and filtered, took 1mL of the filtrate in a 50mL beaker, evaporated the filtrate to dryness in a constant temperature water bath at 60°C, added 0.02mol hydrochloric acid to dilute 4 times, Filtrate with a 0.22 μm filter membrane and measure with a Hitachi L-8800 automatic amino acid analyzer. The crude protei...

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Abstract

The invention belongs to the technical field of biology, and discloses pichia anomala as well as a fermentation culture and application thereof. The pichia anomala is pichia anomala AR2016, CCTCC NO:M2017594. The fermentation culture medium provided by the invention is used for liquid state fermentation; the biomass of the fermentation liquid is 15g / L; the yield of the Killer toxin is 5.86mg / L; the yield of beta-1,3-glucanase is 964 U / L; the yield of phytase is 1675 U / L. The pichia anomala provided by the invention has the effects of inhibiting dysentery colibacillus, salmonella pullorum, sensitive C type candida albicans, actinobacillus pleuropneumoniae, sarcina lutea and staphylococcus aureus and resisting gastric acid and bile salts. Therefore the pichia anomala provided by the invention can be used as feed additives for livestock, poultry and aquatic animals; wide application prospects are realized.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to Pichia anomaly and its fermentation culture and application. technical background [0002] Pichia anomaly is a basidiomycete, the colony is milky white, it is a simple pseudohyphae, the cell shape is round, and it is a vegetative somatic cell that buds and reproduces. Pichia anomaly has anti-Candida albicans, gastric acid and bile salt resistance, and its liquid and solid fermentation products also contain killing factors, β-1,3-glucanase and phytase. β-1,3-glucanase can catalyze the hydrolysis of β-glucan in the cell wall of grain. Phytase can hydrolyze phosphate residues from phytic acid, destroy the strong affinity of phytic acid to mineral elements, increase the nutritional value of mineral elements, and improve the nutritional value of plant feed. Pichia anomaly produces phagocytosis factor, β-1,3-glucanase and phytase by fermentation. The culture cycle is shor...

Claims

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Application Information

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Patent Type & Authority Applications(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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