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Pichia kudriavzevii strain (Pichia kudriavzevii) capable of producing beta-phenethyl alcohol, culture method and application of pichia kudriavzevii strain

A technology of yeast strain and phenethyl alcohol is applied in the field of microorganisms to achieve the effects of optimizing medium components and culture conditions, improving yield and wide tolerance range of pH value.

Active Publication Date: 2020-08-14
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the problem that natural green β-phenylethanol cannot meet the market demand, and the practical application of increasing the content of β-phenylethanol in traditional brewed food liquor and improving the quality of liquor, the purpose of the present invention is to provide a high-yield β-phenylethanol newly isolated from liquor koji. -Phenylethyl alcohol strain, named as YF1702, and provide YF1702 production medium, culture method and application of β-phenylethanol

Method used

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  • Pichia kudriavzevii strain (Pichia kudriavzevii) capable of producing beta-phenethyl alcohol, culture method and application of pichia kudriavzevii strain
  • Pichia kudriavzevii strain (Pichia kudriavzevii) capable of producing beta-phenethyl alcohol, culture method and application of pichia kudriavzevii strain
  • Pichia kudriavzevii strain (Pichia kudriavzevii) capable of producing beta-phenethyl alcohol, culture method and application of pichia kudriavzevii strain

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Experimental program
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Embodiment 1

[0053] The isolation of embodiment 1 YF1702 bacterial strain

[0054] Collect Daqu (the starting material for liquor production), distiller's grains (fermented grains for liquor distillation) or soil from different liquor production enterprises to obtain yeast and store them in glycerol tubes. Take 100 μl of yeast preserved in glycerol tubes, Add 50mLYPD medium under aseptic operation, culture at 28°C and 180r / min for 24h, and activate the strain. Take 100 μL of the activated yeast, add it into the liquid fermentation medium under aseptic operation, and cultivate it at 30°C and 180r / min for 48h. The formula of the liquid fermentation medium is: glucose 100g / L, magnesium sulfate 0.2g / L, potassium dihydrogen phosphate 5g / L, L-phenylalanine 10g / L, and the initial pH value is the natural pH. Screen the strains that can transform L-phenylalanine into β-phenylethanol, and measure the content of β-phenylethanol, and finally obtain a strain numbered YF1702 in Luzhou Laojiao distiller...

Embodiment 2

[0056] Example 2 Identification of YF1702 bacterial strain

[0057] 1. Morphological characteristics: In order to identify the yeast strains involved, the following morphological characteristics were observed.

[0058] YPD medium bacterium colony morphology and cell observation: the bacterial strain pure culture obtained in embodiment 1 is inoculated on the YPD solid medium, observes its morphological characteristic after 48h ( figure 1 ), the results are shown in Table 1, and observed under an optical microscope after being magnified 400 times, the results are as follows figure 2 , fusiform, budding, without mycelium.

[0059] Table 1 Morphological characteristics of strain YF1702

[0060]

[0061] 2. Physiological and biochemical identification: The following physiological and biochemical determinations were carried out on the YF1702 strain:

[0062] (1) Sugar fermentation experiment: The tested sugars include the following sugar sources: D-galactose, L-sorbose, D-ara...

Embodiment 3

[0086] Growth characteristics of embodiment 3 YF1702 bacterial strain

[0087] The growth characteristic of YF1702 bacterial strain, comprises following experiment:

[0088] 1. Sugar tolerance: Based on YPD medium, prepare glucose and sucrose concentrations of 30%, 40%, 50%, 60%, 70%, 80%, and 90%, and distribute them in 250mL conical flasks. Make three parallels, culture at 28°C and 180r / min for 24h with 2% inoculum size, to measure OD 600 As a result of high glucose tolerance, results such as Figure 4 , 5 shown by Figure 4 It can be seen that the glucose tolerance of the strain exceeds 70%, which is determined by Figure 5 It can be seen that the sucrose tolerance of the strain is more than 80%, and it has better sugar tolerance.

[0089] 2. Growth pH: Based on YPD medium, prepare liquid medium with pH of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 and 13, and inoculate activated Test the strain, cultivate it at 28°C and 180r / min for 24h, and measure the OD 600 , the resu...

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Abstract

The invention discloses a pichia kudriavzevii strain (Pichia kudriavzevii) capable of producing beta-phenethyl alcohol, a culture method and application of the pichia kudriavzevii strain. The strain is preserved in the China General Microbiological Culture Collection Center on April 26, 2020, and the preservation number of the strain is CGMCC No.19729; the strain has good sugar tolerance, ethanoltolerance and phenethyl alcohol tolerance, and is wide in growth pH and temperature range; the strain can be used for biologically converting L-phenylalanine to prepare beta-phenylethanol, and detection shows that the yield of the beta-phenylethanol of the strain can reach 5.09 g / L, so that the application of the strain in industrial production of beta-phenylethanol is facilitated, and the strainhas the advantages of mild reaction conditions, no seasonal influence, relatively short period, relatively low cost, no pollution and the like.

Description

technical field [0001] The invention belongs to the field of microbes, and in particular relates to a strain of Pichia pastoris producing beta-phenylethanol, a culture method and application thereof. Background technique [0002] Baijiu is a drink with a long history in China. Various ethnic groups brew a variety of flavors according to their preferences, such as eight flavors: light-flavored, strong-flavored, sauce-flavored, sesame-flavored, and medicinal-flavored. In recent years, due to the development of technology and people's high requirements for food safety flavor, more and more researchers have begun to study the flavor substances in liquor. According to literature reports, 1737 kinds of trace flavor volatile substances and more than 200 kinds of alcohols have been detected in liquor. Among them, β-phenylethanol is one of the constituents in the aroma system of Daqu, and it is an important taste substance in liquor, which can be measured by solid phase microextract...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12P7/22C12R1/84
CPCC12P7/22C12N1/16C12R2001/84C12N1/165
Inventor 范光森孙宝国李秀婷成柳洁富志磊
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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