Yarrowia lipolytica and method for producing gamma-decalactone by using same

A technology of Yarrowia lipolytica and decalactone, which is applied in the direction of microbial-based methods, biochemical equipment and methods, and bulk chemical production, to achieve the effects of increasing yield, good economic value and application potential, and improving yield

Inactive Publication Date: 2016-04-20
西藏天虹科技股份有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It can be seen that the production of GDL by chemical synthesis involves the application of organic so

Method used

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  • Yarrowia lipolytica and method for producing gamma-decalactone by using same

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Embodiment 1

[0036] The method of the present invention comprises the following steps:

[0037] 1. Fermentation strain: Yarrowia Lipolytica (Yarrowia Lipolytica) C85 screened by nitrosoguanidine (NTG) mutagenesis # .

[0038] Nitrosoguanidine (NTG) mutagenesis: Centrifuge the seed medium in the logarithmic growth phase of the medium (6000r / min5min), wash with 20mL phosphate buffer each time, centrifuge twice, combine, draw 20ml of bacterial suspension into a In the small Erlenmeyer flask with glass beads wrapped in a black bag, add 0.6ml NTG (preparation method: add 2.5mg NTG, distilled water to 10ml, final concentration is 0.25mg / ml). The colonies with better growth were selected and subjected to shake flask fermentation experiments, and were repeatedly screened by NTG treatment to obtain C85 # , after fermentation, the lactone output can reach more than 8.7g / L.

[0039] 2. Medium:

[0040] (1) Slant medium: wort slant (recipe: malt extract powder 90-120g / L, yeast powder 5-10g / L, agar...

Embodiment 2

[0051] Take the primary seeds in a 500mL Erlenmeyer flask, fill with 150-200mL seed culture medium, inoculum size 10%-15%, culture on a shaker at 25°C, 175r / min for 48h, initial pH 6.0, to obtain secondary seeds.

[0052] Inoculation of large-volume fermentation culture in a 50L fermenter: transfer the secondary seeds into a 50L fermenter (with 30L sterilized fermentation medium inside) according to the inoculum amount of 2%, and use 5% castor methanolic acid methyl ester respectively At 64h, 76h, 92h, and 102h, feed was fed in batches, adding 3% glucose and 2% methyl ricinoleate in a mass-volume ratio to the fermentation medium. The fermentation temperature was adjusted to 27° C. and the fermentation time was 288 hours. The measured concentration of γ-decalactone was 87.53%, the yield was 14.52 g / L, and the conversion rate was 49.34%.

[0053] Among them, the seed medium contains: glucose 15g / L, yeast extract 2.5g / L, peptone 3.5g / L, magnesium sulfate 2.4g / L, potassium dihydro...

Embodiment 3

[0056] Take the primary seeds in a 500mL Erlenmeyer flask, fill with 150-200mL seed culture medium, inoculum size 10%-15%, culture on a shaker at 25°C, 175r / min for 48h, initial pH 6.0, to obtain secondary seeds.

[0057] Inoculation of large-volume fermentation culture in a 50L fermenter: transfer the secondary seeds into a 50L fermenter (with 30L sterilized fermentation medium inside) according to the inoculum amount of 2%, and use 5% castor methanolic acid methyl ester respectively At 64h, 76h, 92h, and 102h, feed was fed in batches, adding 4% glucose and 7% methyl ricinoleate in a mass-volume ratio to the fermentation medium. The fermentation temperature was adjusted to 27° C. and the fermentation time was 288 hours. The measured concentration of γ-decalactone was 85.1%, the yield was 14.9 g / L, and the conversion rate was 49.2%. Among them, the seed medium contains: glucose 18g / L, yeast extract 7.5g / L, peptone 7.5g / L, magnesium sulfate 5g / L, potassium dihydrogen phosphate ...

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Abstract

The invention discloses a Yarrowia lipolytica C85#. The Yarrowia lipolytica C85# is collected by General Microorganism Center of China Committee for Culture Collection of Microorganisms; the collection number is CGMCC No.11736, and the collection time is November 25th, 2015. The invention also discloses a method for producing gamma-decalactone by using the Yarrowia lipolytica. The method optimizes the high-yield strain fermentation technique, determines the fermentation technological parameters, and enhances the yield of the gamma-decalactone.

Description

technical field [0001] The invention belongs to the technical field of industrial biology, relates to a Yarrowia lipolytica, and also relates to a method for preparing gamma-decalactone by fermentation of the Yarrowia lipolytica. Background technique [0002] γ-Decalactone (γ-Decalactone, GDL) is a lactone fragrance substance, it is a ten-carbon compound containing a five-membered lactone ring, the molecular formula is C 10 h 18 o 2 , the molecular weight is 170.25. GDL is a colorless liquid with coconut, peach, citrus, coconut aroma, boiling point 153°C / 2000Pa or l14-116°C / 66.7Pa, refractive index (nD21.5) 1.4610, slightly soluble in water, easily soluble in organic solvents. It is widely used in the preparation of flavors such as cream, peach, citrus and coconut, and is one of the most demanded products in spices. GDL naturally exists in peaches, strawberries, coconuts, mangoes, apricots, beer, and rum. It has vinegar, fruity, creamy, peach, and coconut aromas. It also...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12P17/04C12R1/645
CPCC12P17/04C12N1/145C12R2001/645Y02P20/54
Inventor 张春颖
Owner 西藏天虹科技股份有限责任公司
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