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Fermentation method for natural perfume gamma-decalactone

A technology of natural flavor and fermentation method, applied in the directions of microorganism-based methods, fermentation, biochemical equipment and methods, etc., can solve the problem of incomplete conversion of castor oil, long period of biotransformation of castor oil, and efficiency of γ-decalactone biosynthesis. Low problems, to achieve the effect of lifting inhibition, improving conversion rate and increasing yield

Inactive Publication Date: 2014-06-11
SHANGHAI INST OF TECH
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Problems solved by technology

[0005] The purpose of the present invention is in order to solve above-mentioned biofermentation production gamma-decalactone process, the biotransformation period of castor oil is long, castor oil transforms incompletely and finally Due to technical problems such as low biosynthetic efficiency of the product γ-decalactone, a fermentation method of natural flavor γ-decalactone is provided, that is, using Saccharomyces cerevisiae (Saccharomyces cerevisiae) as the strain, adding Exogenous porcine pancreatic lipase carries out the conversion mode of coupling reaction, to improve the lipolysis process of castor oil, thereby reduces the biotransformation cycle of castor oil, the biotransformation cycle of castor oil of the present invention is only 12~30h, castor oil The conversion rate can reach 95.0%, and the biosynthesis efficiency of the final product γ-decalactone can reach 94.9%

Method used

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  • Fermentation method for natural perfume gamma-decalactone

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preparation example Construction

[0040] (2) Preparation of seed liquid for fermentation

[0041] Use the sterilized inoculation shovel to inoculate the slant bacteria with a size of 1cm*2cm into a 500mL shaker flask with 50mL seed medium, place it on a shaking table and control the speed at 200-300r / min, and the temperature is 28 Cultivate at ℃ for 24h to obtain seed solution;

[0042] Described seed culture medium is calculated by every liter, by the castor oil of 1.0g, the glucose of 20g, the (NH 4 ) 2 HPO 4 , 3.2g of KH 2 PO 4 , 1.5g of K 2 HPO 4 ·3H 2 O, 4.0g of MgSO 4 ·7H 2 O, 3.0g of NaCl, 15.0g of yeast extract and the remainder of water, the pH is natural.

[0043] 2. The content of γ-decalactone in the fermented liquid obtained in each embodiment of the present invention adopts the gas phase analysis method, and the instrument used is a gas chromatograph, namely Agilent 6890N, produced by Agilent Technology Co., Ltd. of the United States, and the chromatographic column is HP-5 Column (30mX...

Embodiment 1

[0051] Shake flask fermentation medium is calculated per liter, and it consists of 20g of castor oil, 10g of glucose, 10.0g of yeast extract, 0.1g of (NH 4 ) 2 HPO 4 , 0.5g of KH 2 PO 4 , 2.5g of K 2 HPO 4 ·3H 2 O, 1.2g of MgSO 4 ·7H 2 O, 7.0g of NaCl, 0.1g of porcine pancreatic lipase and the rest of the water, the pH is natural.

[0052] After sterilizing the above-mentioned shake flask containing the shake flask fermentation medium at 121°C for 20 minutes, insert 2% seed solution in a sterile room at a temperature of 25°C, the shake flask is 250mL, and the liquid volume is 20%, and then carried out fermentation culture on a shaker at a controlled temperature of 32°C and a rotational speed of 200r / min, and the fermentation ended after 30 hours of constant temperature cultivation.

[0053] After the fermentation is finished, it is calculated according to the volume ratio, that is, the ratio of fermented liquid: butyl acetate is 1:1. After adding butyl acetate to the ...

Embodiment 2

[0056] Shake flask fermentation medium is calculated per liter, and it consists of castor oil of 10g, glucose of 20g, yeast extract of 20g, (NH 4 ) 2 HPO 4 , 3.0g of KH 2 PO 4 , 1.2g of K 2 HPO 4 ·3H 2 O, 3.0g of MgSO 4 ·7H 2 O, the NaCl of 3.5g, the lipase of 2.0g and the water of remainder, pH is natural.

[0057] After sterilizing the above-mentioned shake flask containing the shake flask fermentation medium at 121°C for 20 minutes, insert 2% seed solution in a sterile room at a temperature of 25°C, the shake flask is 250 mL, and the liquid The amount is 20%, and then on a shaker, the temperature is controlled at 36°C, the rotation speed is 300r / min, and the fermentation is completed after 30 hours of constant temperature cultivation.

[0058] After the fermentation is finished, it is calculated according to the volume ratio, that is, the ratio of fermented liquid: butyl acetate is 1:1. After adding butyl acetate to the obtained fermented liquid, rotate and shake f...

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Abstract

The invention discloses a fermentation method for a natural perfume gamma-decalactone, Namely, under the conditions that ((i)saccharomyces cerevisiae ( / i)) CGMCCNo.2.630 is used as a strain, and castor oil is used as a substrate, porcine pancreatic lipase is added, a natural perfume gamma-decalactone is prepared through a batch fermentation or fed-batch fermentation method in an aerobic environment. The fermentation method for a natural perfume gamma-decalactone disclosed by the invention reduces the cycle of transforming castor oil into natural gamma-decalactone and improves the ratio of transforming castor oil into natural gamma-decalactone and the yield of final products gamma-decalactone, wherein the biological transformation cycle of castor oil is 12-30h, the transformation rate of castor oil can reach 95%, and the biosynthesis efficiency of gamma-decalactone can reach 94.9%, especially in a 5L fermentation tank, the final yield of gamma-decalactone prepared through batch fermentation can reach 4.81g / L, and the final yield of gamma-decalactone prepared through fed-batch fermentation can reach 6.50g / L.

Description

technical field [0001] The present invention relates to a fermentation method of natural flavor gamma-decalactone, in particular to a bacterial strain purchased by our laboratory which adopts metabolizable castor oil as gamma-decalactone. The China General Microbiological Culture Collection Center (CGMCC) is the Saccharomyces cerevisiae ( Saccharomyces cerevisiae ), the CGMCC number is cgmcc 2.630, and it is fermented through the synergistic effect with external porcine pancreatic lipase, a method for preparing natural flavor γ-decalactone by biological fermentation. Background technique [0002] γ-decalactone (γ-decalactone, GDL) has a boiling point of 281°C at standard atmospheric pressure. It is a colorless to pale yellow clear liquid with a pleasant peach and coconut aroma, and is easily soluble in organic solvents. γ-decalactone is not only found in peaches, coconuts, mangoes and other fruits, but also one of the characteristic taste substances of dairy products. So fa...

Claims

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Application Information

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IPC IPC(8): C12P17/04C12R1/865
Inventor 荣绍丰曹顺利蔡宝国管世敏李茜茜
Owner SHANGHAI INST OF TECH
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