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 (Saccharomyce

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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