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Phaffia rhodozyma with high output of lycopene and production method of lycopene

A technology for lycopene and a production method is applied in the field of Phaffia rhodozyma strain with high lycopene production and lycopene production, and can solve the problems of low stability coefficient of engineering bacteria, complicated operation process and the like

Active Publication Date: 2019-05-28
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the stability coefficient of engineering bacteria is low, and there may be hidden safety hazards, and there are still many problems to be solved before industrial production
At present, the main strain of industrial production of lycopene is Blakeslea trispora, but most of the products are mixed carotenoids, and the operation process is complicated.

Method used

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  • Phaffia rhodozyma with high output of lycopene and production method of lycopene
  • Phaffia rhodozyma with high output of lycopene and production method of lycopene

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Embodiment 1 Preparation of Phaffia rhodozyme strain

[0022] Collect the orchard soil of Jilin Agricultural University, inoculate the orchard soil on a YPD medium plate, and cultivate at a constant temperature of 20-28°C for 48-96 hours, select red colonies and inoculate them in the YEPD medium, and carry out 150-270r in a constant temperature shaker at 20-28°C Cultivate for 48-96 hours per minute, measure the lycopene production, and obtain the strain with higher lycopene production as PR106, then carry out bacterial morphology, physiology and biochemistry and 26s rRNA identification, the length of 26s rRNA RNA is 522bp, through phylogeny The tree compares this strain to Phaffia rhodozyma ( Phaffia rhodozyma ), named Phaffia rhodozyma PR106, and was deposited in the China Center for Type Culture Collection on January 24, 2019, at Wuhan University, Wuhan City, Hubei Province, and the preservation number is CCTCC M2019083.

Embodiment 2

[0023] Embodiment 2 strain culture

[0024] Strain activation: Take Phaffia rhodozyme PR106, culture it in a shaker flask, culture it by streaking, and culture it in a biochemical incubator at 20-28°C. After 7 days, select a large and dark single colony and insert it into the liquid medium , the liquid medium was sterilized at 121°C for 15 minutes, the bacteria were cultured at 18-28°C, 150-270r / min for 48-96h, and passed on for three generations according to this procedure;

[0025] Seed culture: the activated strain after subculture, use YEPD liquid medium, the inoculation amount is 1-10%, and cultivate in a shaker for 48-96h under the conditions of 18-28°C and 120-300r / min.

Embodiment 3

[0026] Example 3 Extraction and Purification of Lycopene

[0027] Phaffia rhodozyme PR106 was inoculated into a 100mL Erlenmeyer flask with 30mL of fermentation medium at 10% inoculum, cultured on a shaking table at 20°C and 180r / min for 72h; dissolved oxygen was 40-90%, and lye was added to control , pH6.0-7.5; the fermentation medium can also be replaced by YEPD medium, but considering the production cost and fermentation cycle, the fermentation medium is the most suitable;

[0028] Then carry out cell disruption, take 20 ml of fermentation broth and put it into a 50 ml centrifuge tube, centrifuge at 5000r / min at 4°C for 10 minutes to collect the precipitate to obtain the bacteria, wash with distilled water, centrifuge and discard the supernatant three times to obtain yeast sludge, add 5 mL of pre-prepared 3mol / L hydrochloric acid solution, oscillate evenly, soak for 30 minutes to acidify the yeast cell wall, bathe in boiling water for 4-5 minutes, and then immediately put i...

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Abstract

The invention discloses phaffia rhodozyma. The collection number of the phaffia rhodozyma is CCTCC M 2019083. A production method of the lycopene comprises the following steps of (1) selecting the phaffia rhodozyma, activating, inoculating into a fermenting culture medium, and culturing in a bed shaking way, so as to obtain a fermenting solution; (2) centrifuging the fermenting solution, collecting precipitate to obtain strain, washing by water, centrifuging, removing supernatant, adding 3mol / L of hydrochloric acid solution, uniformly oscillating, soaking, boiling in water bath, cooling, centrifuging, collecting precipitate, washing by water, centrifuging, and removing supernatant, so as to obtain cell fragments; (3) adding acetone, oscillating and extracting under the light shading condition, centrifuging, and collecting supernatant; (4) relieving pressure and evaporating, adding methanol, obtaining a methanol lycopene solution, transferring into a chromatographic column, eluting by methanol, collecting the eluting solution, relieving pressure and evaporating, so as to obtain the lycopene; preparing the dry strain with lycopene content of 30.32mg / g.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a Phaffia rhodozyme strain with high lycopene production and a lycopene production method. Background technique [0002] Lycopene, also known as ψ-carotene, molecular formula C 40 h 56 , with a relative molecular mass of 536.8, composed of 11 conjugated double bonds and 2 non-conjugated double bonds. It is orange-yellow in a lower concentration solution, usually dark red powder or oily liquid, and pure lycopene is needle-like dark red, insoluble in water, difficult to dissolve in polar organic solvents, soluble in low polar organic solvents Organic solvents, so less polar organic solvents are usually used to extract lycopene. Lycopene is a terpenoid with strong antioxidant activity. It has various physiological functions such as quenching singlet oxygen, scavenging free radicals, and inhibiting lipid peroxidation. Its ability to quench singlet oxygen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12P5/02C12R1/645
Inventor 王玉华张晶代伟长朴春红于寒松刘俊梅
Owner JILIN AGRICULTURAL UNIV
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