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Method for producing aromatic substances by Trichoderma reesei

A technology of Trichoderma reesei and fermentation substrate, applied in the field of producing beta-phenylethanol and/or phenylacetaldehyde, can solve the problems of expensive L-phenylalanine raw material, limited initial raw material of phenylethanol, and high fermentation cost , to achieve the effect of shortening production time, non-toxic growth, and diversifying metabolic capacity

Active Publication Date: 2020-06-26
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The bottleneck mainly exists in that the process of converting L-phenylalanine into β-phenylethanol in yeast depends on the conversion efficiency of this metabolic process in the bacteria, and this process is only available when L-phenylalanine is the only nitrogen source Only when it can happen, the initial raw material for the production of phenylethanol is limited, and the raw material of L-phenylalanine is expensive, and the fermentation cost is relatively high

Method used

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  • Method for producing aromatic substances by Trichoderma reesei
  • Method for producing aromatic substances by Trichoderma reesei
  • Method for producing aromatic substances by Trichoderma reesei

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Trichoderma reesei utilizes bran to produce β-phenylethanol

[0038] Fermentation medium (g / L): bran 60 (Yihai Kerry (Kunshan) Food Industry Co., Ltd.), glucose 1, ammonium sulfate 1.84, yeast extract 0.5 (OXOID, product number LP0021), potassium dihydrogen phosphate 0.2, Magnesium sulfate heptahydrate 0.5, calcium chloride dihydrate 0.03, L-phenylalanine added at 0, 1.5, 3, 4.5, 6 and 7.5 g / L, pH natural.

[0039] The above medium was divided into 250mL shake flasks, 50ml per bottle, and sterilized at 115°C for 20min.

[0040] Strain cultivation: inoculate the spores of nine kinds of Trichoderma reesei in Table 1 on PDA medium respectively, and culture them at 28° C. for 7 days. PDA medium formula: boil 200g potatoes for half an hour, filter through 4 layers of gauze, and make 1 liter. After sterilization at 115°C, add 20 g of glucose.

[0041] Preparation of spore suspension: the Trichoderma reesei strains cultured on the PDA culture plate for 7 days we...

Embodiment 2

[0050] Example 2 Trichoderma reesei utilizes defatted rice bran meal to produce β-phenylethanol

[0051] In order to further investigate the effect of using other cheap carbon sources to cultivate Trichoderma reesei to produce β-phenylethanol, randomly selected strains GIM3.498, GIM3.538, and CBS383.78 were fermented on the medium with defatted rice bran meal as the carbon source.

[0052] Fermentation medium (g / L): defatted rice bran meal 60 (Yihai Kerry (Jiamusi) Grain and Oil Industry Co., Ltd.), glucose 1, ammonium sulfate 1.84, yeast extract (OXOID, product number LP0021) 0.5, potassium dihydrogen phosphate 0.2 , magnesium sulfate heptahydrate 0.5, calcium chloride dihydrate 0.03, L-phenylalanine addition 0, 1.5, 3, 4.5, 6 and 7.5, pH natural.

[0053] The above medium was divided into 250mL shake flasks, 50mL per shake flask, and sterilized at 115°C for 20min.

[0054] The experimental method is the same as in Example 1, except that the culture medium used is different....

Embodiment 3

[0062] Example 3 Fermentation in 7.5L and 30L fermenters using fermentation strain CICC40929

[0063] In combination with Examples 1-2, we proved that CICC40929, BRC31326, GIM3.498, GIM3.538, CBS383.78, CICC40358, CICC40360, CICC41491, and CICC41493 can produce β-phenylethanol at the shake flask level. Through Example 3, we further verified the ability of the Trichoderma reesei strain to produce phenylethanol at the fermenter level, wherein CICC40929 was randomly selected for fermentation in a 7.5-liter tank and a 30-liter tank, respectively. For the 7.5 liter fermenter experiment, the main carbon source used was bran; the 30 liter fermenter used glucose as the carbon source.

[0064] (1) Fermentation of CICC40929 in 7.5L tank

[0065] Fermentation medium

[0066] Fermented seed medium (g / L): malt extract 20, glucose 20, ammonium sulfate 1.84, yeast extract 0.5, potassium dihydrogen phosphate 0.2, magnesium sulfate heptahydrate 0.5, calcium chloride dihydrate 0.03, pH natu...

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Abstract

The invention provides a method for producing beta-phenethyl alcohol and / or phenylacetaldehyde. The method is to provide a cheap carbon source (such as bran and degreased rice bran) to ferment trichoderma reesei to produce a natural aromatic substance, for example beta-phenethyl alcohol and phenylacetaldehyde. The method for producing the beta-phenethyl alcohol, disclosed by the invention, can reduce the production cost, and utilizes the agricultural byproducts such as the bran and the degreased rice bran, so as to reduce the waste treatment cost.

Description

technical field [0001] The present invention relates to a method for producing natural fragrance substances by Trichoderma reesei. Specifically, the present invention relates to a method for producing β-phenylethanol and / or phenylacetaldehyde using Trichoderma reesei. Background technique [0002] β-phenylethanol (2-Phenylethanol, 2-PE for short) is a derivative of aromatic hydrocarbons, which widely exists in nature and was first discovered as a characteristic aroma compound in roses. It is a pleasant rose-scented volatile compound isolated from the essential oils of many flowers such as rose, carnation, hyacinth, Aleppo pine, orange blossom, lily of the valley and geranium, and is It is a widely used fragrance compound in the field of perfume and cosmetics because of its rose scent. [0003] β-Phenethyl alcohol can be converted into phenylethyl acetate with acetic acid (or acetic anhydride). The product has a sweet aroma of rose honey and peach, and can be used to prepar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/22C12P7/24C12R1/885
CPCC12P7/22C12P7/24
Inventor 曾阿娜曾祥康袁永红朱辰冯奇许骏
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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