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Method of using trichoderma reesei to produce aromatic substance

A technology of Trichoderma reesei and fermentation substrate, which is applied in the field of production of β-phenylethanol and/or phenylacetaldehyde, and can solve the problem of limited initial raw materials for phenylethanol, high fermentation costs, and expensive L-phenylalanine raw materials, etc. problem, to achieve the effect of shortening the production time, diversifying the metabolic capacity, and growing without toxicity

Active Publication Date: 2015-04-29
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 of using trichoderma reesei to produce aromatic substance
  • Method of using trichoderma reesei to produce aromatic substance
  • Method of using trichoderma reesei to produce aromatic substance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Trichoderma reesei uses 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, catalog number LP0021), potassium dihydrogen phosphate 0.2, Magnesium sulfate heptahydrate is 0.5, calcium chloride dihydrate is 0.03, and the addition amount of L-phenylalanine is 0, 1.5, 3, 4.5, 6 and 7.5g / L, and the pH is natural.

[0039] The above-mentioned culture medium was divided into 250 mL shake flasks, 50 mL each, and sterilized at 115°C for 20 min.

[0040] Strain culture: 9 spores of Trichoderma reesei in Table 1 were respectively inoculated on PDA medium and cultured at 28°C for 7 days. PDA medium formula: boil 200g potatoes for half an hour, filter with 4 layers of gauze, and make 1 liter. After sterilization at 115°C, add 20 g of glucose.

[0041] Spore suspension preparation: The Trichoderma reesei strains cultured on the PDA culture plate fo...

Embodiment 2

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

[0051] In order to further investigate the effect of using other cheap carbon sources to culture Trichoderma reesei to produce β-phenethyl alcohol, strains GIM3.498, GIM3.538, and CBS383.78 were randomly selected for fermentation on a medium using defatted rice bran meal as a 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, catalog number LP0021) 0.5, potassium dihydrogen phosphate 0.2 , Magnesium sulfate heptahydrate 0.5, Calcium chloride dihydrate 0.03, L-phenylalanine addition amount 0, 1.5, 3, 4.5, 6 and 7.5, pH natural.

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

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

Embodiment 3

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

[0063] Combined with Examples 1-2, we proved that CICC40929, BRC31326, GIM3.498, GIM3.538, CBS383.78, CICC40358, CICC40360, CICC41491, 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, where CICC40929 was randomly selected for fermentation in a 7.5-liter tank and a 30-liter tank. For the 7.5-liter fermentation tank experiment, the main carbon source used was bran; the 30-liter fermentation tank used glucose as the carbon source.

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

[0065] Fermentation medium

[0066] Fermentation 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, natural pH

[0...

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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 flavor 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 is widely present in nature and was first discovered as a characteristic aroma compound in roses. It is a pleasant volatile substance with a rose fragrance, which can be separated from the essential oils of many kinds of flowers (such as rose, carnation, hyacinth, Aleppo pine, orange blossom, lily of the valley and geranium). Because of its rose fragrance, it has become a fragrance compound widely used in the field of perfume and cosmetics. [0003] β-Phenylethanol can be converted into phenethyl acetate with acetic acid (or acetic anhydride). The product has the sweet aroma of rose honey and ...

Claims

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Application Information

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Patent Type & Authority Applications(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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